Vikas Dighe | NIRRH - Academia.edu (original) (raw)
Papers by Vikas Dighe
VETERINARY SCIENCE RESEARCH JOURNAL, 2014
The aim of the study was to assess the usefulness of 16S rRNA gene in identification of Brucella ... more The aim of the study was to assess the usefulness of 16S rRNA gene in identification of Brucella spp. and phylogenetic study. For identification purposes, a 1477-bp fragment of 16S rRNA gene of 16 isolates and 3 control reference strains was amplified and 3 isolates with B. abortus S19 sequenced. The obtained sequences were submitted to GenBank for species identification. Sequence analysis of a 1477-bp 16S rRNA fragment allows the identification of Brucella spp. However, for discrimination of closely related species sequencing of the entire 16S rRNA gene, additional sequencing of the 23S rRNA gene or sequencing of the 16S-23S rRNA intergenic spacer is essential.
Bioorganic & Medicinal Chemistry Letters
In our previous study, we had identified a 9-mer peptide (FSHβ (89-97)) derived from seat belt lo... more In our previous study, we had identified a 9-mer peptide (FSHβ (89-97)) derived from seat belt loop of human FSHβ and demonstrated its ability to function as FSHR antagonist in vivo. Structure analysis revealed that the four central residues 91STDC94 within this peptide may not be critical for receptor binding. In the present study, 91STDC94 residues were substituted with alanine to generate ΔFSHβ 89-97(91STDC94/AAAA) peptide. Analogous to the parent peptide, ΔFSHβ 89-97(91STDC94/AAAA) peptide inhibited binding of iodinated FSH to rat FSHR and reduced FSH-induced cAMP production. The peptide could impede granulosa cell proliferation leading to reduction in FSH-mediated ovarian weight gain in immature female rats. In these rats, peptide administration further downregulated androgen receptor and estrogen receptor-alpha expression and upregulated estrogen receptor-beta expression. The results indicate that substitution of 91STDC94 with alanine did not significantly alter FSHR antagonist activity of FSHβ (89-97) peptide implying that these residues are not critical for FSH-FSHR interaction and can be replaced with non-peptidic moieties for development of more potent peptidomimetics.
Indian Journal of Medical Research
Background & objectives: Due to limited information available on the frequency and spectrum o... more Background & objectives: Due to limited information available on the frequency and spectrum of cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene mutations in congenital bilateral absence of vas deferens (CBAVD) in Indian population, it is difficult to provide accurate genetic counselling to couples. The present study was undertaken to investigate the spectrum and frequency of CFTR gene mutations in Indian men with CBAVD and to determine the female CF carrier status. Methods: Direct DNA sequencing of the CFTR gene was carried out in eighty CBAVD men, their female partners and fifty controls from the general population. Pathological significance of the identified novel CFTR gene variants was carried out using in silico tools. Appropriate genetic counselling was provided to the couples prior to intracytoplasmic sperm injection (ICSI). Results: A significant association was observed for CFTR gene variants in Indian CBAVD men versus controls (odds ratio: 12.1; 95% confidence interval: 4.8-30.4; P<0.0001). A total of 20 CFTR gene variants were identified in 53 CBAVD men. Eight novel missense CFTR gene variants (L214V, A238P, E379V, L578I, F587L, L926W, R1325K and R1453Q); two novel splice-site gene variants (c.1-30C>G and IVS1+2T>G) and ten previously reported mutations (R75Q, c.1210-12[5], F508del, A309G, R334W, I444T, R668C, R709X, A1285V and Q1352H) were detected in CBAVD men. The novel and reported CFTR gene mutations were L926W (2.5%, P=0.26), R1453Q (2.5%, P=0.26), F508del (8.75%, P=0.03) and c.1210-12[5] (42.5%, P=0.002). A total of 13 (16.2%) female partners were found to be a CF carrier. Nine couples had a risk of transmitting mutant CFTR allele to the offspring. Interpretation & conclusions: The heterogeneous spectrum of CFTR gene in Indian population suggests the necessity of screening CBAVD men and female partners for accurate genetic counselling prior to undergoing ICSI.
Systems Biology in Reproductive Medicine
Over the recent years, FSHR has become an important target for development of fertility regulatin... more Over the recent years, FSHR has become an important target for development of fertility regulating agents, as impairment of FSH-FSHR interaction can lead to subfertility or infertility. In our previous study, we identified a 9-mer peptide (FSHβ (89-97)) that exhibited FSHR antagonist activity. The histopathological and biochemical observations indicated, in addition to FSHR antagonism, a striking resemblance to a PCOS-like state. These observations led us to hypothesize that use of FSHR antagonists can trigger a PCOS-like state. In the present study, to validate this hypothesis, we performed qRT-PCR validation using ovarian tissue samples from our previous study. Expression of three genes known to be differentially expressed in PCOS was evaluated and found to be similar to the PCOS state. To further test the hypothesis, theoretical simulations were carried out by using the human menstrual cycle model available in the literature. Model simulations for FSHR antagonism were indicative of increased testosterone levels, increased ratio of luteinizing hormone/follicle stimulating hormone, and stockpiling of secondary follicles, which are typical characteristics of PCOS. The findings of this study will be relevant while reviewing the utility of FSHR antagonists for fertility regulation and reproductive medicine.Abbreviations: FSH: Follicle-stimulating hormone; FSHR: Follicle-stimulating hormone receptor; cAMP: Cyclic adenosine 3'5' monophosphate; PKA: Protein kinase A; PI3K: Phosphoinositide 3-kinase; PKB: protein kinase B; ERK1/2: Extracellular signal-regulated protein kinase 1/2; MAPK: Mitogen-activated protein kinases; T: testosterone; E2: estradiol; PCOS: Polycystic ovarian syndrome; LH: luteinizing hormone; Lhcgr: luteinizing hormone/choriogonadotropin receptor; CYP17A1: cytochrome P450 family 17 subfamily A member 1; Inhba: inhibin subunit beta A; qRT-PCR: Real-Time quantitative reverse transcription polymerase chain reaction; FSHβ: Follicle-stimulating hormone β subunit; Ct: Cycle threshold; Rn18s: Rattus norvegicus 18S ribosomal RNA.
Biochimica et Biophysica Acta (BBA) - Biomembranes
Interaction of follicle stimulating hormone (FSH) with its cognate receptor (FSHR) is critical fo... more Interaction of follicle stimulating hormone (FSH) with its cognate receptor (FSHR) is critical for maintaining reproductive health. FSHR has a large extracellular domain (ECD), composed of leucine rich repeats (LRRs) and hinge region, a transmembrane domain (TMD) and a short C-terminal domain (CTD). In this study, we have identified a short peptidic stretch in the hinge region (hFSHR(271-275)), through extensive computational modeling, docking and MD simulations, that is capable of independently interacting with the extracellular loops of FSHR(TMD). In vitro studies revealed that FSHR(271-275) peptide increased binding of [125I]-FSH to rat Fshr as well as FSH-induced cAMP production. Administration of FSHR(271-275) peptide in immature female rats significantly increased FSH-mediated ovarian weight gain and promoted granulosa cell proliferation. In summary, the results demonstrate that the synthetic peptide corresponding to amino acids 271-275 of hFSHR-hinge region stimulates FSH-FSHR interaction and behaves as positive allosteric modulator of FSHR. The study also lends evidence to the existing proposition that hinge region maintains the receptor in an inactive conformation in the absence of its ligand by engaging in intramolecular interactions with extracellular loops of TMD.
Materials Today Communications
Abstract Critical size calvarial bone defects and their repair is the major challenge in orthopae... more Abstract Critical size calvarial bone defects and their repair is the major challenge in orthopaedic surgery. Bone tissue engineering using varied combinations of scaffolds, stem cells, and growth factors is an emerging choice of treatment for variety of bone defects. Present study aimed to compare and evaluate the potential of poly e-caprolactone (PCL), PCL- graphene oxide (GO), PCL-GO-Cissus quadrangularis (CQ), and human umbilical cord-derived mesenchymal stem cells (hUCMSCs) seeded PCL (PCL- hUCMSCs), PCL-GO-CQ-hUCMSCs scaffolds to heal critical size calvarial bone defect in the rat models. Healthy adult Wister female rats (N = 30) with average body weight of 350 ± 30 g were divided to 6 groups with five animals in each group. Single critical size calvarial defects of 8 mm were created in the skull of each rat and same were treated with respective scaffolds. The outcome of the treatments was evaluated at 6 weeks and 12 weeks in terms of weight, haematological parameters and biochemical parameters for biocompatibility. New bone regeneration/healing was analysed using digital radiography and micro-computed tomography. Quality of bone formed was analysed by bone mineral density. Histological analysis of the bone tissues was performed using an optical microscope. All the scaffolds were biocompatible and there was no adverse effect of these scaffolds on animals. Higher bone regeneration was observed after 12 weeks of transplantation than 6 weeks. However, PCL-GO-CQ-hUCMSCs scaffolds exhibited highest bone regeneration 12 weeks post-transplantation. The unique combination of PCL-GO-CQ-hUCMSCs scaffold proved to be beneficial in bone regeneration by aiding nearly complete healing of defect site. Further studies with PCL-GO-CQ-hUCMSCs could pave the way for human clinical trials.
Parkinson’s disease (PD) ranks as second most prevalent neurodegenerative disorder but is devoid ... more Parkinson’s disease (PD) ranks as second most prevalent neurodegenerative disorder but is devoid of neuroprotective treatment. Approaches with disease modifying ability with symptomatic relief has become an utmost necessity. Further multifactorial nature of PD presents challenges for efficacy evaluation of any potential test compound. The stated study makes an attempt to address these issues by employing a rotenone induced PD model involving a bilateral intranigral stereotactic rotenone injection for evaluation of the neuroprotective efficacy of Daidzein (DZ). DZ a soy isoflavone, is known for its various health benefits viz. immunomodulation, cardiovascular effects etc. In this study, animals after intranigral rotenone (12 μg) injection, were treated with DZ at a dose of 5, 10 and 20 mg/kg for 30 days. The neurobehavioural evaluation comprised of Rota-rod, Open field and Barnes maze test. The biochemical analysis constituting oxidative stress (Reduced glutathione, superoxide dismut...
Food and Chemical Toxicology
Several scientific reports suggest perturbed reproductive and developmental defects associated wi... more Several scientific reports suggest perturbed reproductive and developmental defects associated with environmental exposure to Atrazine (ATR). ATR has been associated with altered endocrine and reproductive functioning in-vivo exposed during the critical window of development. Thus, the present study investigates the effect of ATR exposure on F1-F2 male progeny exposed through gestation and lactation. F0 dams administered with ATR at doses 2, 10, 70, and 100 mg/kg b. wt/day from gestation day 6 to postnatal day 21. The F1 male rats were monitored for sexual maturation and subjected to fertility assessment on PND75. Delayed testicular descent was observed in 10, 70, and 100 mg/kg b. wt/day ATR dose with significantly lower serum testosterone, sperm count, and motility with testicular defects in F1 male. Expression of Androgen receptor (AR), Estrogen receptors (ER α and ER β), StAR, Aromatase, and INSL-3 were upregulated at all doses indicating estrogenic/anti-androgenic activity of ATR. Fertility assessment revealed subfertility in F1 males with high (%) pre- and post-implantation loss at 10, 70, and 100 mg/kg b. wt/day dose as compared to control. Further, F2 fetuses exhibited congenital disabilities viz. decreased weight, crown-rump length, and anogenital distance with several other morphological deformities. To conclude, ATR exerted estrogenic and/or anti-androgenic activity with fetotoxic effects through the male germline.
Journal of Food Biochemistry
Journal of Human Reproductive Sciences
Aims and Objectives: To study development of neo-vagina by metaplastic conversion of peritoneum, ... more Aims and Objectives: To study development of neo-vagina by metaplastic conversion of peritoneum, To identify translational Stemness markers using NANOG/OCT4/SOX2 from serial neo-vaginal mRNA, cDNA and to study role of WNT and HOXA genes in patients undergoing vaginoplasty. Material and Methods: 75 MRKH Syndrome women underwent laparoscopic peritoneal vaginoplasty (LPV). Two patients underwent serial neo-vaginal biopsies on day 0, 7-9, 12-14, 21 and 33. Fifteen MRKHS and twelve controls were subjected for neo-vaginal biopsy to detect genes upregulation. Remaining patients were evaluated anatomically and functionally. Results: The translational stemness markers NANOG, OCT4 and SOX2 responsible for neo-vaginal formation were identified. Their appearance, concentration at different stages of conversion were demonstrated. The neo-vagina has shown up-regulation of these translational stemness markers. The study demonstrates expression of the specific genes (WNT4, WNT5A and WNT7A) and their role in formation of the neo-vagina. In the subjects stemness markers (NANOG, OCT4 and SOX2) appeared from day 9 to 14 of the neo-vaginal biopsies and after achieving the peak declined later. Genetic analysis showed low values in HOXA 9,10,11,13 and up-regulation of WNT 4A,5A,7 genes in neo-vagina. Conclusions: Study shows peritoneal metaplastic conversion to normal vagina, identified the translational stemness markers and genes responsible. The neo-vagina has shown up-regulation of these genes. The study demonstrates expression of the specific genes (WNT4, WNT5A and WNT7A) and their role in formation of the neo-vagina. Furthering this research, activating these genes may lead to treatment of developmental defects of Mullerian duct, obviating the need of transplant.
Peptides
FSH-FSHR interaction is critical for folliculogenesis as well as progression of several cancers. ... more FSH-FSHR interaction is critical for folliculogenesis as well as progression of several cancers. FSHR peptidic antagonists can circumvent the side effects associated with currently available steroidal contraceptives. The present study aims to identify the shortest peptidic stretch of FSH that can exhibit FSHR antagonistic activity. Based on homology and structural analysis of FSH-FSHR complex (PDB ID: 4AY9), a minimal continuous stretch within FSHβ seat-belt loop (FSHβ (89-97)) was identified to be crucial for FSH-FSHR interaction. Binding affinity and activity of FSHβ (89-97) peptide was evaluated using in silico, in vitro and in vivo methods. The peptide could significantly inhibit binding of [ 125 I] FSH to rat FSHR as well as FSH-induced cAMP production. In vivo administration of this peptide resulted in reduced ovarian weight in immature Holtzman female rats. The peptide inhibited transition of follicles from pre-antral to antral stage as well as progression of granulosa cells beyond G0/G1 phase. Administration of FSHβ (89-97) peptide in adult female rats inhibited conversion of testosterone to estradiol and could significantly retard folliculogenesis. In summary, FSHβ (89-97) peptide is a potential candidate for further optimization for use as fertility regulator or theranostic agent in cancer therapy.
European Journal of Drug Metabolism and Pharmacokinetics
An innovative intranasal aqua-triggered in-situ (ATIS) gel is a polymer-free in-situ gelling micr... more An innovative intranasal aqua-triggered in-situ (ATIS) gel is a polymer-free in-situ gelling microemulsion which gels instantaneously on contact with minute quantities of water to form a mucoadhesive gel. The objective of the study was to develop ATIS diazepam (ATIS-diazepam) as an alternative to the injection for epileptic emergencies and evaluate its brain uptake and nose-to-brain targeting efficiency in rats. ATIS-diazepam (1 mg/100 µL) was prepared and characterized for in vitro formulation characteristics. An LC–MS/MS method was developed and validated for the bioanalysis of diazepam. In vivo studies for pharmacokinetics, brain uptake and nasal irritation of intranasal ATIS-diazepam were conducted in rats. Brain uptake was investigated with brain microdialysis, a highly sensitive technique enabling quantification of free drug, which correlates to efficacy. ATIS-diazepam exhibited globule size < 200 nm, low viscosity, negative zeta potential and good stability. A significant increase in mucoadhesion was exhibited by ATIS-diazepam following the addition of a small quantity of water. ATIS-diazepam showed burst release in pH 6.4 with 50% diazepam release in ~ 10 min, which was sustained over 1 h. The absolute bioavailability was ~ 50% with both intranasal free-diazepam and ATIS-diazepam. Intranasal administration of ATIS-diazepam revealed immediate absorption with rapid and high brain extracellular fluid concentration compared to intravenous free-diazepam solution. The estimated direct transport potential and drug targeting efficiency of intranasal ATIS-diazepam was significantly higher (2-fold) than intranasal free-diazepam solution, which was attributed to the mucoadhesive and microemulsion properties of ATIS-diazepam. The nasal irritation study revealed the safety of ATIS-diazepam compared to free-diazepam solution. Intranasal ATIS-diazepam showed promise of higher direct nose-to-brain targeting, better safety and hence has an immense implication in the treatment of epileptic emergencies.
Brain Research Bulletin
Parkinson's disease (PD) is an age associated, progressive, second most common neurodegenerat... more Parkinson's disease (PD) is an age associated, progressive, second most common neurodegenerative disease, caused due to degeneration of dopaminergic neurons in the substantia nigra (SN). Various studies imply mitochondrial dysfunction, oxidative stress, altered degradation of misfolded proteins in PD pathogenesis. Ursolic acid (UA), a natural pentacyclic triterpenoid carboxylic acid, is reported to possess a number of biological activities viz. anti-oxidant, anti-inflammatory. The focus of our study was to assess the neuroprotective potential of UA against the rotenone induced pathophysiological alterations. In this study rats were subjected to stereotaxic bilateral injection of rotenone (12 µg/µl) in SN. Further, they were treated per-orally with UA (5 and 10 mg/kg) for 30 days. During the study, neurobehavioural studies comprising Rota-rod, open field and Barnes maze test (BMT) were conducted. At the end of 30 days, the antioxidant (Reduced glutathione, superoxide dismutase, catalase and lipid peroxidation), inflammatory (TNF-α) parameters, immunohistochemistry for TH positive neurons, Glial Fibrillary Acidic Protein (GFAP) and mitochondrial complex I and mitochondrial biogenesis (MB) were assessed. The results showed a significant amelioration in the motor deficits by UA which can be attributed to the protection of TH positive neurons degeneration. A significant improvement in the cognitive function due to UA was observed in BMT. Biochemically, the oxidative stress and inflammation triggered by rotenone was significantly diminished by UA. UA also significantly obviated the complex I inhibition and promoted MB. The preliminary results thus firmly advocate the neuroprotective potential of UA to prevent rotenone induced neurotoxicity in rats.
Veterinary Medicine and Science
AI is the predominant methodology used in bovine reproduction around the world. Main objective of... more AI is the predominant methodology used in bovine reproduction around the world. Main objective of the dairy farm is to produce one calf per cow annually. An important element of economical calf production is top quality bull semen for successful fertilization and improved herd health (Swanson & Herman, 1940). Generation of reactive oxygen species (ROS) throughout freeze thaw cycle accompanied by low antioxidant levels in seminal plasma and in
Environmental Pollution
Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol, TCS), is a broad-spectrum antimicrobial agen... more Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol, TCS), is a broad-spectrum antimicrobial agent and extensively used in household and daily daycare products. Recently, several reports have demonstrated the endocrine disruptive action of TCS to alter the testicular steroidogenesis. However, the gestational and lactational effects of TCS exposure on F1 offspring has not been studied. Present study aimed to investigate the effect of gestational and lactational exposure to TCS on F1 male progeny and its effect on fertility. Pregnant dams (F0) were administered with different doses of TCS (0.1, 4, 40 and 150 mg/kg b. wt./day) and Diethylstilbestrol (1 μg/kg b. wt./day), as a positive control daily by subcutaneous injection during Gestation Day 6 to Postnatal Day 21. Delayed testicular descent was observed at 150 mg/kg b. wt./day dose group. Dose-dependent decrease in testosterone level, sperm count and motility was observed. Significantly decreased expression of steroid hormone receptors (AR, ERα and ERβ), StAR and aromatase were observed in F1 male rats; indicating its prolonged effect on spermatogenesis and steroidogenesis in adulthood and poor development in F2 fetuses. Further, gestational and lactational exposure to TCS has negative impact on the fertility of F1 male rats. The F1 male rats were found sub-fertile with increased (%) pre- and post-implantation loss (at 40 and 150 mg/kg b.wt./day dose) with a simultaneous decrease in litter size. The significant decrease in mean fetal weight and crown-rump length (CRL) of F2 fetuses were observed at 0.1, 4, 40 and 150 dose groups indicating impaired development of F2 fetuses caused by TCS exposure. Present study emphasizes for the first time that TCS exposure during the vulnerable developmental time point (gestation and lactation) adversely affects reproductive functions and fertility of F1 male rats, which were transmitted to F2 generations leading to reduced CRLs and weights of F2 fetuses.
Environmental Pollution
Parabens are class of preservatives used in vast majority of commercial products, and a potential... more Parabens are class of preservatives used in vast majority of commercial products, and a potential Endocrine Disrupting Chemical (EDC). The present study was undertaken to delineate the effects of n-butylparaben on F1 male progeny exposed maternally through gestation and lactation via subcutaneous route. The F0 dams were given subcutaneous injections of n-butylparaben from gestation day (GD) 6 to postnatal day (PND) 21 with doses of 10, 100, 1000 mg/kg Bw/day in corn oil. The F1 male rats were monitored for pubertal development and sexual maturation; these were sacrificed on PND 30, 45 and 75. On PND 75, these F1 male rats were subjected for fertility assessment with unexposed female rats. A delayed testicular descent at 100 and 1000 mg/kg Bw dose and delayed preputial separation at 10 mg/kg Bw dose was observed in exposed F1 male rats. Decreased sperm count, motility and Daily Sperm Production was observed at 100 mg/kg Bw dose at PND 75. Interestingly, the sperm transit time in the epididymis was accelerated at this dose. Significant perturbed testicular expression of steroid receptors (ERα and β, AR), INSL3 and StAR genes with increased T and LH levels indicates direct effect on spermatogenesis and steroidogenesis. These F1 generation adult rats were sub-fertile with increased (%) pre- and post-implantation loss at 100 and 1000 mg/kg Bw/day dose. This is the first report on n-butylparaben highlighting the involvement of testicular leydig cells with accelerated sperm transit time leading to reduced fertility in the maternally exposed F1 male rats through estrogenic/anti-androgenic action.
Frontiers in Pharmacology
Epilepsy is a brain disorder characterized by sudden recurrent seizures. Considering the fact tha... more Epilepsy is a brain disorder characterized by sudden recurrent seizures. Considering the fact that epileptogenesis is a process that affects the quality of life, our goal is to delay the process of epileptogenesis and to increase the latency of epileptic attacks, offering better quality of life to patients. Traditional system of medicines has a promise in some of the medicines, which have been used for the treatment of epilepsy. One such medicinal plant is Eclipta alba (EA). According to Ayurvedic philosophy, the juice of leaves of EA is pounded with garlic and pepper for the treatment of epilepsy. Taking clue from the Ayurvedic system of medicines, we formulated coumarin fraction of EA, namely, coumarin nasal formulation (CNF) for its nasal delivery. CNF was analyzed by using high performance liquid chromatography (HPLC) and ultraviolet absorption spectroscopy for its drug content determination. In vitro drug release studies were performed in simulated nasal electrolyte solution (SNES) maintaining constant pH of 5.5 at 37 • C. Irritation by CNF was evaluated using hen's egg test chorioallantoic membrane (HET-CAM) assay. Formulation was found to be non-irritant in HET-CAM assay. CNF was further assessed in vivo by measuring the progress and attainment of pentylenetetrazole (PTZ) kindling in mice. Neuronal changes were assessed by hematoxylin and eosin (H&E) and Nissl staining technique. Glial fibrillary acidic protein (GFAP) a neuroinflammatory marker and tumor necrosis factor alpha (TNF-α) an inflammatory marker were also measured. CNF (10 mg/kg, nasal route) when given as a pretreatment lowered seizure score and delayed the progression of seizure similar to diazepam. CNF decreased the PTZ induced oxidative damage, TNF-α as well as GFAP levels in the midbrain tissue particularly in hippocampus region. The results suggest that CNF may be a promising therapeutic approach to offer protection from sudden recurrent seizures alone or in combination with current drugs in management of epilepsy.
Naunyn-Schmiedeberg's Archives of Pharmacology
Alzheimer's disease (AD) is the leading neurodegenerative disorder with extracellular sen... more Alzheimer's disease (AD) is the leading neurodegenerative disorder with extracellular senile plaques and neurofibrillary tangles as the major hallmarks. The objective was to evaluate the effect of phloretin in a chronic model of sporadic AD by injecting aggregated form of Aβ25-35 peptide sequence intracerebroventricularly (icv) in Wistar rats. To achieve this, male Wistar rats were injected with aggregated Aβ25-35 peptide icv, followed by 21 days phloretin (2.5 mg/kg, 5 mg/kg) administration after recovery period. Barnes maze and elevated plus maze along with the biochemical estimation of antioxidant enzymes activities were conducted. The hippocampus region of the rat brains were stained with Congo red and Nissl stain. TNF-α was estimated in the brain homogenates using the ELISA assay. In this study, phloretin improved the spatial memory formation and retention in Barnes maze test. Additionally, phloretin alleviated the antioxidant defense biomarkers and thereby reduced oxidative stress, decreased TNF-α-mediated neuroinflammation. Furthermore, phloretin treatment showed decreased amyloid beta accumulation in the CA1 region and less number of pyknotic nuclei in the dentate gyrus of the Aβ25-35-injected rat brains. The above experimental findings evinced the promising role of phloretin in Aβ25-35-injected rats and which further envisage its potential to be explored in the treatment of AD.
VETERINARY SCIENCE RESEARCH JOURNAL, 2014
The aim of the study was to assess the usefulness of 16S rRNA gene in identification of Brucella ... more The aim of the study was to assess the usefulness of 16S rRNA gene in identification of Brucella spp. and phylogenetic study. For identification purposes, a 1477-bp fragment of 16S rRNA gene of 16 isolates and 3 control reference strains was amplified and 3 isolates with B. abortus S19 sequenced. The obtained sequences were submitted to GenBank for species identification. Sequence analysis of a 1477-bp 16S rRNA fragment allows the identification of Brucella spp. However, for discrimination of closely related species sequencing of the entire 16S rRNA gene, additional sequencing of the 23S rRNA gene or sequencing of the 16S-23S rRNA intergenic spacer is essential.
Bioorganic & Medicinal Chemistry Letters
In our previous study, we had identified a 9-mer peptide (FSHβ (89-97)) derived from seat belt lo... more In our previous study, we had identified a 9-mer peptide (FSHβ (89-97)) derived from seat belt loop of human FSHβ and demonstrated its ability to function as FSHR antagonist in vivo. Structure analysis revealed that the four central residues 91STDC94 within this peptide may not be critical for receptor binding. In the present study, 91STDC94 residues were substituted with alanine to generate ΔFSHβ 89-97(91STDC94/AAAA) peptide. Analogous to the parent peptide, ΔFSHβ 89-97(91STDC94/AAAA) peptide inhibited binding of iodinated FSH to rat FSHR and reduced FSH-induced cAMP production. The peptide could impede granulosa cell proliferation leading to reduction in FSH-mediated ovarian weight gain in immature female rats. In these rats, peptide administration further downregulated androgen receptor and estrogen receptor-alpha expression and upregulated estrogen receptor-beta expression. The results indicate that substitution of 91STDC94 with alanine did not significantly alter FSHR antagonist activity of FSHβ (89-97) peptide implying that these residues are not critical for FSH-FSHR interaction and can be replaced with non-peptidic moieties for development of more potent peptidomimetics.
Indian Journal of Medical Research
Background & objectives: Due to limited information available on the frequency and spectrum o... more Background & objectives: Due to limited information available on the frequency and spectrum of cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene mutations in congenital bilateral absence of vas deferens (CBAVD) in Indian population, it is difficult to provide accurate genetic counselling to couples. The present study was undertaken to investigate the spectrum and frequency of CFTR gene mutations in Indian men with CBAVD and to determine the female CF carrier status. Methods: Direct DNA sequencing of the CFTR gene was carried out in eighty CBAVD men, their female partners and fifty controls from the general population. Pathological significance of the identified novel CFTR gene variants was carried out using in silico tools. Appropriate genetic counselling was provided to the couples prior to intracytoplasmic sperm injection (ICSI). Results: A significant association was observed for CFTR gene variants in Indian CBAVD men versus controls (odds ratio: 12.1; 95% confidence interval: 4.8-30.4; P<0.0001). A total of 20 CFTR gene variants were identified in 53 CBAVD men. Eight novel missense CFTR gene variants (L214V, A238P, E379V, L578I, F587L, L926W, R1325K and R1453Q); two novel splice-site gene variants (c.1-30C>G and IVS1+2T>G) and ten previously reported mutations (R75Q, c.1210-12[5], F508del, A309G, R334W, I444T, R668C, R709X, A1285V and Q1352H) were detected in CBAVD men. The novel and reported CFTR gene mutations were L926W (2.5%, P=0.26), R1453Q (2.5%, P=0.26), F508del (8.75%, P=0.03) and c.1210-12[5] (42.5%, P=0.002). A total of 13 (16.2%) female partners were found to be a CF carrier. Nine couples had a risk of transmitting mutant CFTR allele to the offspring. Interpretation & conclusions: The heterogeneous spectrum of CFTR gene in Indian population suggests the necessity of screening CBAVD men and female partners for accurate genetic counselling prior to undergoing ICSI.
Systems Biology in Reproductive Medicine
Over the recent years, FSHR has become an important target for development of fertility regulatin... more Over the recent years, FSHR has become an important target for development of fertility regulating agents, as impairment of FSH-FSHR interaction can lead to subfertility or infertility. In our previous study, we identified a 9-mer peptide (FSHβ (89-97)) that exhibited FSHR antagonist activity. The histopathological and biochemical observations indicated, in addition to FSHR antagonism, a striking resemblance to a PCOS-like state. These observations led us to hypothesize that use of FSHR antagonists can trigger a PCOS-like state. In the present study, to validate this hypothesis, we performed qRT-PCR validation using ovarian tissue samples from our previous study. Expression of three genes known to be differentially expressed in PCOS was evaluated and found to be similar to the PCOS state. To further test the hypothesis, theoretical simulations were carried out by using the human menstrual cycle model available in the literature. Model simulations for FSHR antagonism were indicative of increased testosterone levels, increased ratio of luteinizing hormone/follicle stimulating hormone, and stockpiling of secondary follicles, which are typical characteristics of PCOS. The findings of this study will be relevant while reviewing the utility of FSHR antagonists for fertility regulation and reproductive medicine.Abbreviations: FSH: Follicle-stimulating hormone; FSHR: Follicle-stimulating hormone receptor; cAMP: Cyclic adenosine 3'5' monophosphate; PKA: Protein kinase A; PI3K: Phosphoinositide 3-kinase; PKB: protein kinase B; ERK1/2: Extracellular signal-regulated protein kinase 1/2; MAPK: Mitogen-activated protein kinases; T: testosterone; E2: estradiol; PCOS: Polycystic ovarian syndrome; LH: luteinizing hormone; Lhcgr: luteinizing hormone/choriogonadotropin receptor; CYP17A1: cytochrome P450 family 17 subfamily A member 1; Inhba: inhibin subunit beta A; qRT-PCR: Real-Time quantitative reverse transcription polymerase chain reaction; FSHβ: Follicle-stimulating hormone β subunit; Ct: Cycle threshold; Rn18s: Rattus norvegicus 18S ribosomal RNA.
Biochimica et Biophysica Acta (BBA) - Biomembranes
Interaction of follicle stimulating hormone (FSH) with its cognate receptor (FSHR) is critical fo... more Interaction of follicle stimulating hormone (FSH) with its cognate receptor (FSHR) is critical for maintaining reproductive health. FSHR has a large extracellular domain (ECD), composed of leucine rich repeats (LRRs) and hinge region, a transmembrane domain (TMD) and a short C-terminal domain (CTD). In this study, we have identified a short peptidic stretch in the hinge region (hFSHR(271-275)), through extensive computational modeling, docking and MD simulations, that is capable of independently interacting with the extracellular loops of FSHR(TMD). In vitro studies revealed that FSHR(271-275) peptide increased binding of [125I]-FSH to rat Fshr as well as FSH-induced cAMP production. Administration of FSHR(271-275) peptide in immature female rats significantly increased FSH-mediated ovarian weight gain and promoted granulosa cell proliferation. In summary, the results demonstrate that the synthetic peptide corresponding to amino acids 271-275 of hFSHR-hinge region stimulates FSH-FSHR interaction and behaves as positive allosteric modulator of FSHR. The study also lends evidence to the existing proposition that hinge region maintains the receptor in an inactive conformation in the absence of its ligand by engaging in intramolecular interactions with extracellular loops of TMD.
Materials Today Communications
Abstract Critical size calvarial bone defects and their repair is the major challenge in orthopae... more Abstract Critical size calvarial bone defects and their repair is the major challenge in orthopaedic surgery. Bone tissue engineering using varied combinations of scaffolds, stem cells, and growth factors is an emerging choice of treatment for variety of bone defects. Present study aimed to compare and evaluate the potential of poly e-caprolactone (PCL), PCL- graphene oxide (GO), PCL-GO-Cissus quadrangularis (CQ), and human umbilical cord-derived mesenchymal stem cells (hUCMSCs) seeded PCL (PCL- hUCMSCs), PCL-GO-CQ-hUCMSCs scaffolds to heal critical size calvarial bone defect in the rat models. Healthy adult Wister female rats (N = 30) with average body weight of 350 ± 30 g were divided to 6 groups with five animals in each group. Single critical size calvarial defects of 8 mm were created in the skull of each rat and same were treated with respective scaffolds. The outcome of the treatments was evaluated at 6 weeks and 12 weeks in terms of weight, haematological parameters and biochemical parameters for biocompatibility. New bone regeneration/healing was analysed using digital radiography and micro-computed tomography. Quality of bone formed was analysed by bone mineral density. Histological analysis of the bone tissues was performed using an optical microscope. All the scaffolds were biocompatible and there was no adverse effect of these scaffolds on animals. Higher bone regeneration was observed after 12 weeks of transplantation than 6 weeks. However, PCL-GO-CQ-hUCMSCs scaffolds exhibited highest bone regeneration 12 weeks post-transplantation. The unique combination of PCL-GO-CQ-hUCMSCs scaffold proved to be beneficial in bone regeneration by aiding nearly complete healing of defect site. Further studies with PCL-GO-CQ-hUCMSCs could pave the way for human clinical trials.
Parkinson’s disease (PD) ranks as second most prevalent neurodegenerative disorder but is devoid ... more Parkinson’s disease (PD) ranks as second most prevalent neurodegenerative disorder but is devoid of neuroprotective treatment. Approaches with disease modifying ability with symptomatic relief has become an utmost necessity. Further multifactorial nature of PD presents challenges for efficacy evaluation of any potential test compound. The stated study makes an attempt to address these issues by employing a rotenone induced PD model involving a bilateral intranigral stereotactic rotenone injection for evaluation of the neuroprotective efficacy of Daidzein (DZ). DZ a soy isoflavone, is known for its various health benefits viz. immunomodulation, cardiovascular effects etc. In this study, animals after intranigral rotenone (12 μg) injection, were treated with DZ at a dose of 5, 10 and 20 mg/kg for 30 days. The neurobehavioural evaluation comprised of Rota-rod, Open field and Barnes maze test. The biochemical analysis constituting oxidative stress (Reduced glutathione, superoxide dismut...
Food and Chemical Toxicology
Several scientific reports suggest perturbed reproductive and developmental defects associated wi... more Several scientific reports suggest perturbed reproductive and developmental defects associated with environmental exposure to Atrazine (ATR). ATR has been associated with altered endocrine and reproductive functioning in-vivo exposed during the critical window of development. Thus, the present study investigates the effect of ATR exposure on F1-F2 male progeny exposed through gestation and lactation. F0 dams administered with ATR at doses 2, 10, 70, and 100 mg/kg b. wt/day from gestation day 6 to postnatal day 21. The F1 male rats were monitored for sexual maturation and subjected to fertility assessment on PND75. Delayed testicular descent was observed in 10, 70, and 100 mg/kg b. wt/day ATR dose with significantly lower serum testosterone, sperm count, and motility with testicular defects in F1 male. Expression of Androgen receptor (AR), Estrogen receptors (ER α and ER β), StAR, Aromatase, and INSL-3 were upregulated at all doses indicating estrogenic/anti-androgenic activity of ATR. Fertility assessment revealed subfertility in F1 males with high (%) pre- and post-implantation loss at 10, 70, and 100 mg/kg b. wt/day dose as compared to control. Further, F2 fetuses exhibited congenital disabilities viz. decreased weight, crown-rump length, and anogenital distance with several other morphological deformities. To conclude, ATR exerted estrogenic and/or anti-androgenic activity with fetotoxic effects through the male germline.
Journal of Food Biochemistry
Journal of Human Reproductive Sciences
Aims and Objectives: To study development of neo-vagina by metaplastic conversion of peritoneum, ... more Aims and Objectives: To study development of neo-vagina by metaplastic conversion of peritoneum, To identify translational Stemness markers using NANOG/OCT4/SOX2 from serial neo-vaginal mRNA, cDNA and to study role of WNT and HOXA genes in patients undergoing vaginoplasty. Material and Methods: 75 MRKH Syndrome women underwent laparoscopic peritoneal vaginoplasty (LPV). Two patients underwent serial neo-vaginal biopsies on day 0, 7-9, 12-14, 21 and 33. Fifteen MRKHS and twelve controls were subjected for neo-vaginal biopsy to detect genes upregulation. Remaining patients were evaluated anatomically and functionally. Results: The translational stemness markers NANOG, OCT4 and SOX2 responsible for neo-vaginal formation were identified. Their appearance, concentration at different stages of conversion were demonstrated. The neo-vagina has shown up-regulation of these translational stemness markers. The study demonstrates expression of the specific genes (WNT4, WNT5A and WNT7A) and their role in formation of the neo-vagina. In the subjects stemness markers (NANOG, OCT4 and SOX2) appeared from day 9 to 14 of the neo-vaginal biopsies and after achieving the peak declined later. Genetic analysis showed low values in HOXA 9,10,11,13 and up-regulation of WNT 4A,5A,7 genes in neo-vagina. Conclusions: Study shows peritoneal metaplastic conversion to normal vagina, identified the translational stemness markers and genes responsible. The neo-vagina has shown up-regulation of these genes. The study demonstrates expression of the specific genes (WNT4, WNT5A and WNT7A) and their role in formation of the neo-vagina. Furthering this research, activating these genes may lead to treatment of developmental defects of Mullerian duct, obviating the need of transplant.
Peptides
FSH-FSHR interaction is critical for folliculogenesis as well as progression of several cancers. ... more FSH-FSHR interaction is critical for folliculogenesis as well as progression of several cancers. FSHR peptidic antagonists can circumvent the side effects associated with currently available steroidal contraceptives. The present study aims to identify the shortest peptidic stretch of FSH that can exhibit FSHR antagonistic activity. Based on homology and structural analysis of FSH-FSHR complex (PDB ID: 4AY9), a minimal continuous stretch within FSHβ seat-belt loop (FSHβ (89-97)) was identified to be crucial for FSH-FSHR interaction. Binding affinity and activity of FSHβ (89-97) peptide was evaluated using in silico, in vitro and in vivo methods. The peptide could significantly inhibit binding of [ 125 I] FSH to rat FSHR as well as FSH-induced cAMP production. In vivo administration of this peptide resulted in reduced ovarian weight in immature Holtzman female rats. The peptide inhibited transition of follicles from pre-antral to antral stage as well as progression of granulosa cells beyond G0/G1 phase. Administration of FSHβ (89-97) peptide in adult female rats inhibited conversion of testosterone to estradiol and could significantly retard folliculogenesis. In summary, FSHβ (89-97) peptide is a potential candidate for further optimization for use as fertility regulator or theranostic agent in cancer therapy.
European Journal of Drug Metabolism and Pharmacokinetics
An innovative intranasal aqua-triggered in-situ (ATIS) gel is a polymer-free in-situ gelling micr... more An innovative intranasal aqua-triggered in-situ (ATIS) gel is a polymer-free in-situ gelling microemulsion which gels instantaneously on contact with minute quantities of water to form a mucoadhesive gel. The objective of the study was to develop ATIS diazepam (ATIS-diazepam) as an alternative to the injection for epileptic emergencies and evaluate its brain uptake and nose-to-brain targeting efficiency in rats. ATIS-diazepam (1 mg/100 µL) was prepared and characterized for in vitro formulation characteristics. An LC–MS/MS method was developed and validated for the bioanalysis of diazepam. In vivo studies for pharmacokinetics, brain uptake and nasal irritation of intranasal ATIS-diazepam were conducted in rats. Brain uptake was investigated with brain microdialysis, a highly sensitive technique enabling quantification of free drug, which correlates to efficacy. ATIS-diazepam exhibited globule size < 200 nm, low viscosity, negative zeta potential and good stability. A significant increase in mucoadhesion was exhibited by ATIS-diazepam following the addition of a small quantity of water. ATIS-diazepam showed burst release in pH 6.4 with 50% diazepam release in ~ 10 min, which was sustained over 1 h. The absolute bioavailability was ~ 50% with both intranasal free-diazepam and ATIS-diazepam. Intranasal administration of ATIS-diazepam revealed immediate absorption with rapid and high brain extracellular fluid concentration compared to intravenous free-diazepam solution. The estimated direct transport potential and drug targeting efficiency of intranasal ATIS-diazepam was significantly higher (2-fold) than intranasal free-diazepam solution, which was attributed to the mucoadhesive and microemulsion properties of ATIS-diazepam. The nasal irritation study revealed the safety of ATIS-diazepam compared to free-diazepam solution. Intranasal ATIS-diazepam showed promise of higher direct nose-to-brain targeting, better safety and hence has an immense implication in the treatment of epileptic emergencies.
Brain Research Bulletin
Parkinson's disease (PD) is an age associated, progressive, second most common neurodegenerat... more Parkinson's disease (PD) is an age associated, progressive, second most common neurodegenerative disease, caused due to degeneration of dopaminergic neurons in the substantia nigra (SN). Various studies imply mitochondrial dysfunction, oxidative stress, altered degradation of misfolded proteins in PD pathogenesis. Ursolic acid (UA), a natural pentacyclic triterpenoid carboxylic acid, is reported to possess a number of biological activities viz. anti-oxidant, anti-inflammatory. The focus of our study was to assess the neuroprotective potential of UA against the rotenone induced pathophysiological alterations. In this study rats were subjected to stereotaxic bilateral injection of rotenone (12 µg/µl) in SN. Further, they were treated per-orally with UA (5 and 10 mg/kg) for 30 days. During the study, neurobehavioural studies comprising Rota-rod, open field and Barnes maze test (BMT) were conducted. At the end of 30 days, the antioxidant (Reduced glutathione, superoxide dismutase, catalase and lipid peroxidation), inflammatory (TNF-α) parameters, immunohistochemistry for TH positive neurons, Glial Fibrillary Acidic Protein (GFAP) and mitochondrial complex I and mitochondrial biogenesis (MB) were assessed. The results showed a significant amelioration in the motor deficits by UA which can be attributed to the protection of TH positive neurons degeneration. A significant improvement in the cognitive function due to UA was observed in BMT. Biochemically, the oxidative stress and inflammation triggered by rotenone was significantly diminished by UA. UA also significantly obviated the complex I inhibition and promoted MB. The preliminary results thus firmly advocate the neuroprotective potential of UA to prevent rotenone induced neurotoxicity in rats.
Veterinary Medicine and Science
AI is the predominant methodology used in bovine reproduction around the world. Main objective of... more AI is the predominant methodology used in bovine reproduction around the world. Main objective of the dairy farm is to produce one calf per cow annually. An important element of economical calf production is top quality bull semen for successful fertilization and improved herd health (Swanson & Herman, 1940). Generation of reactive oxygen species (ROS) throughout freeze thaw cycle accompanied by low antioxidant levels in seminal plasma and in
Environmental Pollution
Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol, TCS), is a broad-spectrum antimicrobial agen... more Triclosan (5-chloro-2-(2, 4-dichlorophenoxy) phenol, TCS), is a broad-spectrum antimicrobial agent and extensively used in household and daily daycare products. Recently, several reports have demonstrated the endocrine disruptive action of TCS to alter the testicular steroidogenesis. However, the gestational and lactational effects of TCS exposure on F1 offspring has not been studied. Present study aimed to investigate the effect of gestational and lactational exposure to TCS on F1 male progeny and its effect on fertility. Pregnant dams (F0) were administered with different doses of TCS (0.1, 4, 40 and 150 mg/kg b. wt./day) and Diethylstilbestrol (1 μg/kg b. wt./day), as a positive control daily by subcutaneous injection during Gestation Day 6 to Postnatal Day 21. Delayed testicular descent was observed at 150 mg/kg b. wt./day dose group. Dose-dependent decrease in testosterone level, sperm count and motility was observed. Significantly decreased expression of steroid hormone receptors (AR, ERα and ERβ), StAR and aromatase were observed in F1 male rats; indicating its prolonged effect on spermatogenesis and steroidogenesis in adulthood and poor development in F2 fetuses. Further, gestational and lactational exposure to TCS has negative impact on the fertility of F1 male rats. The F1 male rats were found sub-fertile with increased (%) pre- and post-implantation loss (at 40 and 150 mg/kg b.wt./day dose) with a simultaneous decrease in litter size. The significant decrease in mean fetal weight and crown-rump length (CRL) of F2 fetuses were observed at 0.1, 4, 40 and 150 dose groups indicating impaired development of F2 fetuses caused by TCS exposure. Present study emphasizes for the first time that TCS exposure during the vulnerable developmental time point (gestation and lactation) adversely affects reproductive functions and fertility of F1 male rats, which were transmitted to F2 generations leading to reduced CRLs and weights of F2 fetuses.
Environmental Pollution
Parabens are class of preservatives used in vast majority of commercial products, and a potential... more Parabens are class of preservatives used in vast majority of commercial products, and a potential Endocrine Disrupting Chemical (EDC). The present study was undertaken to delineate the effects of n-butylparaben on F1 male progeny exposed maternally through gestation and lactation via subcutaneous route. The F0 dams were given subcutaneous injections of n-butylparaben from gestation day (GD) 6 to postnatal day (PND) 21 with doses of 10, 100, 1000 mg/kg Bw/day in corn oil. The F1 male rats were monitored for pubertal development and sexual maturation; these were sacrificed on PND 30, 45 and 75. On PND 75, these F1 male rats were subjected for fertility assessment with unexposed female rats. A delayed testicular descent at 100 and 1000 mg/kg Bw dose and delayed preputial separation at 10 mg/kg Bw dose was observed in exposed F1 male rats. Decreased sperm count, motility and Daily Sperm Production was observed at 100 mg/kg Bw dose at PND 75. Interestingly, the sperm transit time in the epididymis was accelerated at this dose. Significant perturbed testicular expression of steroid receptors (ERα and β, AR), INSL3 and StAR genes with increased T and LH levels indicates direct effect on spermatogenesis and steroidogenesis. These F1 generation adult rats were sub-fertile with increased (%) pre- and post-implantation loss at 100 and 1000 mg/kg Bw/day dose. This is the first report on n-butylparaben highlighting the involvement of testicular leydig cells with accelerated sperm transit time leading to reduced fertility in the maternally exposed F1 male rats through estrogenic/anti-androgenic action.
Frontiers in Pharmacology
Epilepsy is a brain disorder characterized by sudden recurrent seizures. Considering the fact tha... more Epilepsy is a brain disorder characterized by sudden recurrent seizures. Considering the fact that epileptogenesis is a process that affects the quality of life, our goal is to delay the process of epileptogenesis and to increase the latency of epileptic attacks, offering better quality of life to patients. Traditional system of medicines has a promise in some of the medicines, which have been used for the treatment of epilepsy. One such medicinal plant is Eclipta alba (EA). According to Ayurvedic philosophy, the juice of leaves of EA is pounded with garlic and pepper for the treatment of epilepsy. Taking clue from the Ayurvedic system of medicines, we formulated coumarin fraction of EA, namely, coumarin nasal formulation (CNF) for its nasal delivery. CNF was analyzed by using high performance liquid chromatography (HPLC) and ultraviolet absorption spectroscopy for its drug content determination. In vitro drug release studies were performed in simulated nasal electrolyte solution (SNES) maintaining constant pH of 5.5 at 37 • C. Irritation by CNF was evaluated using hen's egg test chorioallantoic membrane (HET-CAM) assay. Formulation was found to be non-irritant in HET-CAM assay. CNF was further assessed in vivo by measuring the progress and attainment of pentylenetetrazole (PTZ) kindling in mice. Neuronal changes were assessed by hematoxylin and eosin (H&E) and Nissl staining technique. Glial fibrillary acidic protein (GFAP) a neuroinflammatory marker and tumor necrosis factor alpha (TNF-α) an inflammatory marker were also measured. CNF (10 mg/kg, nasal route) when given as a pretreatment lowered seizure score and delayed the progression of seizure similar to diazepam. CNF decreased the PTZ induced oxidative damage, TNF-α as well as GFAP levels in the midbrain tissue particularly in hippocampus region. The results suggest that CNF may be a promising therapeutic approach to offer protection from sudden recurrent seizures alone or in combination with current drugs in management of epilepsy.
Naunyn-Schmiedeberg's Archives of Pharmacology
Alzheimer's disease (AD) is the leading neurodegenerative disorder with extracellular sen... more Alzheimer's disease (AD) is the leading neurodegenerative disorder with extracellular senile plaques and neurofibrillary tangles as the major hallmarks. The objective was to evaluate the effect of phloretin in a chronic model of sporadic AD by injecting aggregated form of Aβ25-35 peptide sequence intracerebroventricularly (icv) in Wistar rats. To achieve this, male Wistar rats were injected with aggregated Aβ25-35 peptide icv, followed by 21 days phloretin (2.5 mg/kg, 5 mg/kg) administration after recovery period. Barnes maze and elevated plus maze along with the biochemical estimation of antioxidant enzymes activities were conducted. The hippocampus region of the rat brains were stained with Congo red and Nissl stain. TNF-α was estimated in the brain homogenates using the ELISA assay. In this study, phloretin improved the spatial memory formation and retention in Barnes maze test. Additionally, phloretin alleviated the antioxidant defense biomarkers and thereby reduced oxidative stress, decreased TNF-α-mediated neuroinflammation. Furthermore, phloretin treatment showed decreased amyloid beta accumulation in the CA1 region and less number of pyknotic nuclei in the dentate gyrus of the Aβ25-35-injected rat brains. The above experimental findings evinced the promising role of phloretin in Aβ25-35-injected rats and which further envisage its potential to be explored in the treatment of AD.