Tumor necrosis factor alpha and palmitate simulate bovine fatty liver disease in vitro when using abattoir-derived primary bovine hepatocytes isolated by a novel nonperfusion method (original) (raw)
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Invited Review: Pathology, Etiology, Prevention, and Treatment of Fatty Liver in Dairy Cows*
Journal of dairy science, 2004
Fatty liver (i.e., hepatic lipidosis) is a major metabolic disorder of many dairy cows in early lactation and is associated with decreased health status and reproductive performance. In severe cases, milk production and feed intake are decreased. Therefore, a practical preventative or an efficacious treatment of fatty liver could save millions of dollars yearly in treatment, replacement, and production losses for dairy farmers. Fatty liver develops when the hepatic uptake of lipids exceeds the oxidation and secretion of lipids by the liver, which usually is preceded by high concentrations of plasma NEFA mobilized from adipose tissue. Excess lipids are stored as triacylglycerol in the liver and are associated with decreased metabolic functions of the liver. Liver can be categorized into normal liver or mild, moderate, or severe fatty liver; the latter can be subdivided further into nonencephalopathic severe fatty liver and hepatic encephalopathy. Insufficient or unbalanced dietary intake, obesity, and elevated estrogen concentrations are involved in the etiology of fatty liver, which is associated with greater incidence of dystocia, diseases, infections, and inflammations. Because even mild fatty liver is associated with decreased health status and reproductive performance of dairy cows, prevention of fatty liver by supplying cows with sufficient nutrients and a clean and health-promoting environment in the peripartal period would reduce production losses of cows more than would any treatment of fatty liver. This, however, might not be enough for cows that are obese or do not eat well, had calving difficulties or twins, have metabolic or infectious diseases, or are in severe negative energy balance because of high milk production immediately after calving. Potential and commonly used preventatives, as well as treatments, are discussed in the review. Currently, detection of fatty liver is possible only by
Culture of bovine hepatocytes: a non-perfusion technique for cell isolation
Cytotechnology, 2006
In this work we have studied the isolation and culture of mature bovine hepatocytes on plastic dishes without exogenous matrix. The liver has been disaggregated in a collagenase solution instead of undergoing a perfusion step. After a few days in culture, the plates showed several clusters of different cell types. Although the average yield was 1.60-0.57·10 8 viable liver cells per gram of tissue, these cultures were formed by non-parenchymal cells and only very few or none by parenchymal cells. In these cultures, actin structures used as a marker for Stellate (Ito) cells have been visualized by immunocytochemical techniques. In order to increase the proportion of parenchymal cells a centrifugation on Percoll, which separates cell sub-populations, has been introduced. Though the yield was lower than in the previous method, these prepurified cultures were only composed of hepatocytes. It has been shown that these cells exhibited albumin synthesis, which is a specific hepatocytes function. In addition, these cultures were capable of producing metabolites of 7-ethoxycoumarin at a higher rate than non purified cell cultures. Therefore this simplified procedure for the isolation and culture of functional and viable hepatocytes may be applied for in vitro studies in bovine.
Pathological Alterations in Hepatocytes of Dairy Cows With a Tendency to Emaciation and Fattening
Folia Veterinaria
The aim of this study was to demonstrate the histochemical and histopathological alterations in the livers of cows with a tendency to become emaciated (body condition score - BCS1 and 2) and a tendency to become fattened (BCS4 and 5) in comparison to the cows of average body condition (BCS3) presented as a control. The histochemical analysis (PAS reaction) showed that the influence of emaciation and fattening in our study was manifested by a decreased occurrence of glycogen and a decreased level of the PAS-positive matter in the hepatocytes of dairy cows with BCS1, 2, 4 and 5. An abundant accumulation of lipids in the form of large lipid droplets, liposomes and lipoproteins observed in the hepatocytes of emaciated and fattened (BCS1 and 5) cows may be related to moderate-severe steatosis. These observations suggest a relationship between liver steatosis and the occurrence of lipoproteins in cows with a tendency toward emaciation and fattening.
Pakistan Veterinary Journal, 2015
Received: Revised: Accepted: August 30, 2014 November 20, 2014 February 07, 2015 Serious lipid mobilization in adipose tissue of dairy cow is the pathological basis of ketosis and fatty liver. Many Studies demonstrated that pre-adipocytes are unstable and can differentiate into multiple cells in vitro. The aim of this study was to investigate the differentiation process of dairy cow pre-adipocytes and to establish a stable pre-adipocyte induction method. Pre-adipocytes were isolated from dairy cow adipose tissue and were cultured for 15 days. The originally rounded cells converted to a spindle fibroblast-like morphology and accumulated lipids during culturing. The lipid droplets and TG content in the adipocytes gradually increased from 4 to 15 days in culture. The adipocytes reached maximal proliferation after 11 days in culture. Additionally, the expression of peroxisome proliferator-activated receptor γ2 (PPARγ2) and sterol regulatory element binding protein 1c (SREBP-1c) increase...
Exploration of serum sensitive biomarkers of fatty liver in dairy cows
Scientific Reports, 2018
Serum proteins are sensitive with diseases in dairy cows, and some of them could be used as biomarkers for fatty liver. This study aimed to explore serum biomarkers for fatty liver in dairy cows. A total of 28 early lactating dairy cows were chosen from a commercial dairy herds, liver samples were collected for determining concentration of triacylglycerol (TAG), and serum samples were collected for measuring fibroblast growth factor-21 (FGF-21), adiponectin, Lipoprotein-associated phospholipase A2 (LP-PLA2), and hemoglobin (Hb). Dairy cows were divided into fatty liver (liver TAG > 5%, wet weight) and control group (liver TAG < 5%, wet weight). Concentration of FGF-21 was greater in fatty liver cows, while the concentration of LP-PLA2 and Hb was less. The concentration of FGF-21 and total Hb had strong correlation with the liver TAG as well as good prediction power (kappa value = 0.79 and 0.58, respectively). These results suggested that the serum concentration of FGF-21 and t...
Physiological incompatibilities of porcine hepatocytes for clinical liver support
Liver Transplantation, 2006
In fulminant hepatic failure, the use of bioartificial liver support (BAL) with porcine hepatocytes is the subject of a current and controversial debate.1 Specifically, the issue of cross-species physiological incompatibilities has not been addressed so far. We therefore investigated the effects of species-specific cytokines in single and cocultures on hepatocyte function. Hepatocyte cultures were isolated from human resection specimens and from Landrace pigs. Single and cocultures were exposed to porcine and human interleukin (IL)-6 or tumor necrosis factor (TNF)-α. Changes in expression of C-reactive protein (CRP), albumin, CCAAT enhancer binding protein (C/EBP)-α and C/EBP-β and metabolic competence of cultured cells was studied by measuring testosterone metabolite production. After human or porcine IL-6 dosing, CRP was induced up to 100-fold in human hepatocyte cultures, while porcine hepatocytes responded marginally (2- to 5-fold). Treatment with human or porcine IL-6 or TNF-α resulted in reduced albumin production, albeit at different levels when human and porcine hepatocytes were compared (P = 0.001). Unlike human, porcine hepatocytes produced less of 6α-hydroxytestosterone (6α-HT) (P < 0.001) and 7α-HT (P < 0.001) after human or porcine IL-6 dosing and treatment with species-specific TNF-α induced (human hepatocytes) or decreased (porcine hepatocytes) 6β-HT production (P = 0.021). In coculture with free exchange of metabolites, porcine hepatocytes produced less 6α-HT (P = 0.048) and 16α-HT (P = 0.033), whereas after treatment with human IL-6 reduced CRP gene and protein expression was observed with human hepatocytes (P = 0.013). In conclusion, species-specific responses of hepatocytes to cytokines and interactions with xenobiotic metabolites may limit the clinical effectiveness of porcine hepatocytes in BAL. Liver Transpl 12:1832–1840, 2006. © 2006 AASLD.
Hepatocyte apoptosis in dairy cattle during the transition period
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire, 2012
The objective of this study was to investigate hepatocyte apoptosis in dairy cows during the transition period. Four clinically healthy, pregnant dairy cattle were used. The cows had no clinical diseases throughout this study. Blood samples were collected and livers were biopsied from the cows at 3 different times: 3 weeks before expected partition (wk -3); during parturition (wk 0), and 3 weeks (wk +3) after parturition. The damage to deoxyribonucleic acid (DNA) caused by hepatocytes was evaluated by comet assay. The apoptotic features of hepatocytes were examined by immunohistochemistry and electron microscopic analyses. The hepatic triglyceride content markedly increased at wk 0 and wk +3 compared with the values at wk -3. The results of the comet assay showed increases in the mean tail moment values of hepatic cells after parturition in all cows, which suggested increased DNA damage. Histopathologically, the hepatocytes began to contain lipid droplets at wk 0 and were severely o...
Strong relationships between mediators of the acute phase response and fatty liver in dairy cows
Canadian Journal of Animal Science, 2005
The objective of this study was to investigate the relationship between activation of acute phase response and fatty liver in transition dairy cows. Fatty liver was induced in dairy cows by feeding 8 kg of cracked corn 1 mo before the expected day of parturition. Liver and blood samples were obtained at days -4, 3, 8, 12, 14, 22, 27, and 36 postpartum. Cows that developed fatty liver (n = 4) reached peak total lipids in the liver at day 12 postpartum with 11.4% (wet wt.) compared with 6.6% in control cows (n = 4). Cows with fatty liver had greater plasma tumor necrosis factor-alpha (TNF-α), nonesterified fatty acids (NEFA), and lower lactate concentrations than did control cows at day -4. During highest concentrations of total lipids in the liver, for at least one time-point, fatty-liver cows had greater concentrations of plasma serum amyloid A (SAA), haptoglobin, and NEFA and lower concentrations of calcitonin gene-related peptide (CGRP), prostaglandin E2 (PGE2), cortisol, and TNF-...
The functional state of liver cells in dairy cows during transition and lactation
2009
The objective of this study was to determine characteristic blood parameters, i.e. indicators of the functional state of liver in the following groups of cows: in cows (n=15) from day 15 to day 1 prior to parturition, in cows (n=15) from day 1 to day 15 after parturition and in those (n=15) from day 90 to day 100 of lactation. Blood glucose levels were statistically significantly lower in the early lactation cows than in the preparturient cows and those examined during the maximum lactation period, which suggested an increased glucose uptake by the mammary gland at the beginning of lactation and decreased gluconeogenesis in the liver as induced by fatty infiltration and liver cell degeneration. The significantly lower blood triglyceride levels as well as the low total blood cholesterol levels in the postparturient cows reflected their accumulation in liver cells, as opposed to the cows before parturition and those during maximum lactation. Significantly lower blood levels of total p...
Physiological reports, 2017
Human primary hepatocytes are the gold standard for investigating lipid metabolism in nonalcoholic fatty liver disease (NAFLD); however, due to limitations including availability and donor variability, the hepatoma cell lines Huh7 and HepG2 are commonly used. Culturing these cell lines in human serum (HS) has been reported to improve functionality; however, direct comparison of fatty acid (FA) metabolism in response to culturing in HS is lacking. The aim of this study was to compare FA metabolism between HepG2 and Huh7 cells in response to culturing in different sera. Both HepG2 and Huh7 cells were grown in media containing 11 mmol/L glucose and either 2% HS or 10% fetal bovine serum. After 3 days, insulin and insulin-like growth factor-1 signaling were measured. At 7 days, intracellular triacylglycerol (TAG) and media 3-hydroxybutyrate, TAG and apolipoprotein B were measured, as was the FA composition of intracellular TAG and phospholipids. Both cell lines demonstrated higher level...