Department of Reproductive Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China.
Department of Gynecology, Jincheng Maternity and Child Care Hospital, Jincheng 048000, Shanxi Province, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2023 Feb 25;52(1):33-45. doi: 10.3724/zdxbyxb-2022-0456.
To explore the effect and mechanism of Chinese medicine Bushen Huatan formula in treatment of polycystic ovary syndrome (PCOS).
Twenty-four SPF female C57BL/6J mice were randomly divided into 3 groups with 8 animals in each group. Control group was given drinking water ; PCOS was induced by giving letrozole gavage and high-fat diet in model group and treatment group; treatment group received Bushen Huatan formula suspension for 35 d. The sex hormone levels of mice were detected by enzyme-linked immunosorbent assay. Ovary morphology was observed under light microscope after hematoxylin and eosin staining. The feces in the colon of mice were collected, and the gut microbiota was detected by 16S rRNA sequencing. The short chain fatty acids were detected by gas chromatography-mas spectrometry. The expression of peroxisome proliferator activated receptor (PPARγ) was detected by immunohistochemistry. The mRNA expression of mucin-2, occludin-1, tight junction protein zonula occludens 1 (-) and in intestinal epithelium were detected by realtime RT-PCR. The expression of inducible nitric oxide synthase (iNOS) and PPARγ was detected by Western blotting.
Compared with the control group, the body weight, serum levels of follicle stimulating hormone, luteinizing hormone and testosterone in the model group were increased, and serum levels of estradiol were decreased (all <0.01); the ovarian structure under light microscope was consistent with the characteristics of PCOS. Compared with the model group, the serum levels of sex hormone and ovarian structure in treatment group were improved. The overall structure of gut microbiota in PCOS model mice changed. Compared with control group, there were significantly reduced abundance of , and increased abundance of , and inthe model group at phylum level (all <0.05); there were significantly reduced abundance of , and increased abundance of , , and at genus level (all <0.05). The disordered condition of gut microbiota was significantly improved in treatment group. Compared with control group, the contents of acetic acid, propionic acid and butyric acid in feces of model group were significantly decreased (all <0.05); while the contents of propionic acid and butyric acid in treatment group were significantly increased compared with model control group (both <0.05). Compared with control group, the mRNA expression of - and protein expression of iNOS in model group were significantly increased, and the protein expression of PPARγ and the mRNA expressions of - and - were significantly decreased (all <0.05). Compared with model group, the mRNA expression of - and protein expression of iNOS in treatment group were decreased, and the protein expression of PPARγ and the mRNA expressions of mucin-2 and occludin-1 were increased.
PCOS induced by letrozole high-fat diet induces microflora imbalance in mice. Chinese medicine Bushen Huatan formula may increase the level of short chain fatty acid by regulating gut microbiota, thereby activating the intestinal PPARγ pathway and improving intestinal barrier function to act as a cure for PCOS.
探讨补肾化痰方治疗多囊卵巢综合征(PCOS)的作用及机制。
将 24 只 SPF 级 C57BL/6J 雌性小鼠随机分为 3 组,每组 8 只。对照组给予饮用水;模型组和治疗组给予来曲唑灌胃和高脂饮食诱导 PCOS;治疗组给予补肾化痰方混悬液治疗 35 d。采用酶联免疫吸附试验检测小鼠血清性激素水平。苏木精-伊红染色后观察卵巢形态。收集小鼠结肠粪便,采用 16S rRNA 测序检测肠道菌群。采用气相色谱-质谱联用仪检测短链脂肪酸。采用免疫组织化学法检测过氧化物酶体增殖物激活受体(PPARγ)的表达。采用实时 RT-PCR 检测肠上皮细胞中黏蛋白-2(mucin-2)、闭合蛋白-1(occludin-1)、紧密连接蛋白 zonula occludens 1(-)和 的 mRNA 表达。采用 Western blot 检测诱导型一氧化氮合酶(iNOS)和 PPARγ的表达。
与对照组比较,模型组小鼠体重、血清卵泡刺激素、黄体生成素和睾酮水平升高,雌二醇水平降低(均<0.01);卵巢结构符合 PCOS 特征。与模型组比较,治疗组小鼠血清性激素及卵巢结构改善。PCOS 模型小鼠肠道菌群整体结构发生改变。与对照组比较,模型组小鼠厚壁菌门中 、拟杆菌门中 丰度降低,变形菌门中 、放线菌门中 、梭杆菌门中 丰度升高(均<0.05);拟杆菌属、普雷沃氏菌属、双歧杆菌属、罗斯氏菌属、粪杆菌属丰度降低,乳杆菌属、梭菌属、罗氏菌属丰度升高(均<0.05)。治疗组小鼠肠道菌群紊乱状态明显改善。与对照组比较,模型组小鼠粪便中乙酸、丙酸和丁酸含量明显降低(均<0.05);治疗组小鼠粪便中丙酸和丁酸含量较模型对照组升高(均<0.05)。与对照组比较,模型组小鼠 -和 iNOS 蛋白表达增加,PPARγ蛋白表达及 -和 -mRNA 表达降低(均<0.05);与模型组比较,治疗组小鼠 -和 iNOS 蛋白表达降低,PPARγ蛋白表达及 mucin-2 和 occludin-1mRNA 表达升高。
来曲唑高脂饮食诱导的 PCOS 可导致小鼠肠道菌群失衡。补肾化痰方可能通过调节肠道菌群增加短链脂肪酸水平,进而激活肠道 PPARγ 通路,改善肠道屏障功能,从而发挥治疗 PCOS 的作用。