Chitosan alleviated menopausal symptoms and modulated the gut microbiota in estrogen-deficient rats
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ORIGINAL CONTRIBUTION
Chitosan alleviated menopausal symptoms and modulated the gut microbiota in estrogen‑deficient rats Xuangao Wu1 · Min Jung Kim2 · Hye Jeong Yang2 · Sunmin Park1 Received: 11 May 2020 / Accepted: 3 September 2020 © Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Purpose Menopause disturbs energy, glucose, and lipid metabolisms and changes the composition of the gut microbiota, but dietary fibers without phytoestrogens may ameliorate menopausal metabolic disorders. The objective of the present study was to assess whether consuming the prebiotics chitosan and citrus pectin can improve postmenopausal symptoms, possibly by modulating the gut microbiota in ovariectomized (OVX) rats, and the mechanism of action was examined. Methods The OVX rats were given 4.5% cellulose (OVX-Control), chitosan (OVX-Chitosan), or citrus pectin (OVX-Pectin) in a 43% fat diet and the sham rats were given the same diet as the OVX-Control for 12 weeks. Sham-operated rats had the same diet as OVX-Control (Normal-Control). Body-weight, visceral fat mass, tail skin temperature, serum 17β-estradiol, glucose intolerance, and insulin tolerance were determined. Gut microbiota in the fecal samples was measured by NGS and analyzed with PICRUSt2. Short-chain fatty acids (SCFA) and metabolomic characteristics of serum were also measured with UPLC-mass spectrometry. Results Chitosan and citrus pectin were selected because the incubation of rat feces with these two prebiotics in vitro had shown increased butyrate production. OVX-Chitosan reduced the weight, visceral fat content, and tail skin temperature, and OVX-Chitosan and OVX-Pectin improved glucose tolerance, compared to the OVX-Control. Both alleviated dyslipidemia, compared to the OVX-Control. OVX-Chitosan and OVX-Pectin elevated serum propionate and butyrate concentrations but only OVX-Chitosan lowered serum acetate concentrations. In PICRUSt2, chitosan upregulated the functional genes of gut microbiota involved in valine, leucine, and isoleucine biosynthesis, whereas the OVX-Control exhibited significantly upregulated lipopolysaccharide biosynthesis. OVX-Pectin exhibited increased α-diversity in the fecal bacteria. Metabolomic analysis revealed higher serum urate concentrations in the OVX-Control group than the other groups, and serum arginine and leucine concentrations were higher in the OVX-Chitosan group (P 200 base pairs according to the MiSeq SOP using Mothur software (Version 1.43.0). The sequences were divided into the Classification Operation Units (OTUs) according to the 97% sequence similarity, and then the chimera sequences were searched and removed with a 0.97 threshold. The sequences were classified into OTUs to compare according to the Greengenes bacterial reference. The classified OTUs were counted in each sample. Finally, a total of 8,052 and 20,537 OTUs classifications were obtained, with an average of 18,531 ± 727 and 18,483 ± 1251 sequence numbers per sample in vitro and in vivo studies, respectively.
Metabolomics of the serum C
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