Effects of Bu Shen Hua Zhuo formula on the LPS/TLR4 pathway and gut microbiota in rats with letrozole-induced polycystic ovary syndrome - PubMed (original) (raw)

Effects of Bu Shen Hua Zhuo formula on the LPS/TLR4 pathway and gut microbiota in rats with letrozole-induced polycystic ovary syndrome

Yang Wang et al. Front Endocrinol (Lausanne). 2022.

Abstract

Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in gynecology. Traditional Chinese medicine (TCM) is widely used for the treatment of PCOS in China. The Bu Shen Hua Zhuo formula (BSHZF), a TCM decoction, has shown great therapeutic efficacy in clinical practice. However, the mechanism underlying the BSHZF function in PCOS remains unclear. This study aimed to identify the potential mechanisms of action of BSHZF in the treatment of PCOS. PCOS-model rats treated with letrozole were administered different doses of BSHZF, metformin, and 1% carboxymethylcellulose. Serum sex hormones, fasting blood glucose, and fasting insulin levels were measured, and the morphology of the ovaries was observed in each group, including the normal group. The structure and abundance of the gut microbiota in rats were measured using 16S ribosomal RNA gene sequencing. Toll-like receptor 4 (TLR4) and phospho-NF-κB p65 levels in the ovarian tissue of the rats were detected using Western blotting. Furthermore, the levels of lipopolysaccharide (LPS) and inflammatory cytokines TNF-α, IL-6, and IL-8 in the serum of rats were detected by ELISA. The results showed that BSHZF administration was associated with a decrease in body weight, fasting blood glucose, fasting insulin, and testosterone and changes in ovarian morphology in PCOS-model rats. Moreover, BSHZF was associated with an increase in the α-diversity of gut microbiota, decrease in the relative abundance of Firmicutes, and increase in Lactobacillus and short chain fatty acid-producing bacteria (Allobaculum, Bacteroides, Ruminococcaceae_UCG-014). Furthermore, BSHZF may promote carbohydrate and protein metabolism. In addition, BSHZF was associated with a decrease in the serum level of LPS and TLR4 expression, thereby inhibiting the activation of the NF-κB signaling-mediated inflammatory response in ovarian tissue. Therefore, the beneficial effects of BSHZF on PCOS pathogenesis are associated with its ability to normalize gut microbiota function and inhibit PCOS-related inflammation.

Keywords: LPS/TLR4 pathway; gut microbiota; polycystic ovary syndrome; rats; traditional Chinese medicine.

Copyright © 2022 Wang, Xiao, Liu, Tong, Yu, Qi, Bu, Pan and Xing.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1

Figure 1

Associations of the Bu Shen Hua Zhuo formula (BSHZF) with the polycystic ovary syndrome (PCOS) phenotype in rats. (A), Weight curve. (B) Fasting blood glucose (FBG) levels. (C) Fasting insulin (FINS) levels. (D) T levels. * P<0.05, ** P<0.01 versus the model group.

Figure 2

Figure 2

Associations of BSHZF with ovary morphology changes in PCOS-model rats. Hematoxylin and eosin staining of representative ovaries under a 40-fold magnification light microscope. The cystic follicles with monolayer granular cells are indicated by a hashtag (#), while the corpora lutea are indicated by asterisks (*). Scale bar: 180 μm. Control: normal rats, Model: rats administered letrozole, MET: rats administered letrozole and metformin, CHL: rats administered letrozole and a low dose of BSHZF; CHM: rats administered letrozole and a medium dose of BSHZF; CHH: rats administered letrozole and a high dose of BSHZF.

Figure 3

Figure 3

Associations of BSHZF with the regulation of gut microbiota composition in PCOS rats. The feces sample of control 1 was unqualified and could not be used for testing. Furthermore, control 5, model 5, CHM 1, and CHH 6 were eliminated, owing to the overall data deviation. Ultimately, we included four rats in the control group, four in the model group, and five in the CH (CHL, CHM, and CHH) and MET groups. (A), The rarefaction curve. (B), Venn diagram of operational taxonomic units (OTUs). (C), Shannon indexes. (D–G), PCoA plots on OTU (D), phylum (E), family (F), and genus (G) levels. Bacterial composition on phylum (H), family (I), and genus (J) levels. (K) Linear discriminant analysis. (L), relative abundance of Firmicutes. (M) and (N) Relative abundances of bacterial groups with statistically significant differences at the genus level between model and CHH groups and (M) between model and CHM groups (N). (O) and (P) Predicted metabolic pathways by PICRUSt analysis and the statistical difference was estimated by STAMP software between model and CHH groups and (O) between model and CHM groups (P). * P<0.05, ** P<0.01. Control: normal rats, Model: rats administered letrozole, MET: rats administered letrozole and metformin, CHL: rats administered letrozole and a low dose of BSHZF, CHM: rats administered letrozole and a medium dose of BSHZF, CHH: rats administered letrozole and a high dose of BSHZF.

Figure 4

Figure 4

Associations of BSHZF with lipopolysaccharide (LPS) and inflammatory cytokine expression in rats with PCOS. (A) LPS levels in serum measured by ELISA. (B) and (C) Protein expression levels of Toll-like receptor 4 in ovarian tissue detected by Western blot. (D) and (E) Protein expression levels of phospho-NF-κB p65 in ovarian tissue detected by Western blot. (F), IL-18 protein levels in serum measured by ELISA. (G) TNF-α protein levels in serum measured by ELISA. (H), IL-6 protein levels in serum measured by ELISA. * P<0.05, ** P<0.01 versus model group. Control: normal rats, Model: rats administered letrozole, MET: rats administered letrozole and metformin, CHL: rats administered letrozole and a low dose of BSHZF, CHM: rats administered letrozole and a medium dose of BSHZF, CHH: rats administered letrozole and a high dose of BSHZF.

Similar articles

Cited by

References

    1. Goodarzi MO, Dumesic DA, Chazenbalk G, Azziz R. Polycystic ovary syndrome: etiology, pathogenesis and diagnosis. Nat Rev Endocrinol (2011) 7(4):219–31. doi: 10.1038/nrendo.2010.217 - DOI - PubMed
    1. Bruni V, Capozzi A, Lello S. The role of genetics, epigenetics and lifestyle in polycystic ovary syndrome development: the state of the art. Reprod Sci (2021) 29(3):668–79. doi: 10.1007/s43032-021-00515-4 - DOI - PubMed
    1. Ng NYH, Jiang G, Cheung LP, Zhang Y, Tam CHT, Luk AOY, et al. . Progression of glucose intolerance and cardiometabolic risk factors over a decade in Chinese women with polycystic ovary syndrome: A case-control study. PloS Med (2019) 16(10):e1002953. doi: 10.1371/journal.pmed.1002953 - DOI - PMC - PubMed
    1. Joham AE, Kakoly NS, Teede HJ, Earnest A. Incidence and predictors of hypertension in a cohort of Australian women with and without polycystic ovary syndrome. J Clin Endocrinol Metab (2021) 106(6):1585–93. doi: 10.1210/clinem/dgab134 - DOI - PubMed
    1. Løvvik TS, Carlsen SM, Salvesen Ø, Steffensen B, Bixo M, Gómez-Real F, et al. . Use of metformin to treat pregnant women with polycystic ovary syndrome (PregMet2): a randomised, double-blind, placebo-controlled trial. Lancet Diabetes Endocrinol (2019) 7(4):256–66. doi: 10.1016/s2213-8587(19)30002-6 - DOI - PubMed

MeSH terms

Substances

LinkOut - more resources