银甲片通过纠正外周血Th17/Treg平衡改善慢性输卵管炎大鼠不孕

Yinjia Tablets Improve Infertility in Rats with Chronic Salpingitis by Correcting Peripheral Blood Th17/Treg Balance

关键词:  慢性输卵管炎  银甲片  Th17/Treg平衡  受孕率

Key words:  chronic salpingitis  Yinjia tablets  Th17/Treg balance  conception rate

摘要:

目的 探讨银甲片是否通过纠正外周血Th17/Treg平衡改善慢性输卵管炎大鼠不孕。方法 100只雌性Wistar大鼠，其中82只构建慢性输卵管炎大鼠模型，造模成功后，取80只随机分为模型组、左旋氧氟沙星组(20 mg·kg^-1)、银甲片低剂量组(0.38 g·kg^-1·d^-1)、银甲片中剂量组(0.76 g·kg^-1·d^-1)、银甲片高剂量组(1.52 g·kg^-1·d^-1)，每组16只，另取16只设为空白对照组。治疗组分别按不同剂量灌胃给药，模型组与空白对照组灌服等量生理盐水(20 mL·kg^-1·d^-1)，每日1次，灌胃7 d后停药1 d，7 d为1个周期，共灌胃3个周期。给药结束后，观察大鼠受孕情况，采集血样和输卵管组织。采用HE染色观察输卵管组织病理变化；透射电镜观察输卵管组织超微结构变化；ELISA检测血清中TNF-α、IL-6含量；流式细胞术检测外周血Treg、Th17细胞比例；RT-qPCR和Western blotting检测输卵管组织IL-17、IL-10、Foxp3、RORγt mRNA及蛋白表达。结果 与空白对照组相比，模型组大鼠输卵管纤毛、微绒毛稀疏，线粒体肿胀；且TNF-α、IL-6含量、Th17细胞比例及输卵管组织IL-17、RORγt mRNA和蛋白表达明显升高，大鼠受孕率、IL-10、Treg细胞比例及Foxp3 mRNA和蛋白表达明显降低(P<0.05)。与模型组比较，银甲片高剂量组改善大鼠输卵管结构；明显降低模型大鼠TNF-α、IL-6含量、Th17细胞比例及输卵管组织IL-17、RORγt mRNA和蛋白表达；明显升高大鼠受孕率及IL-10、Treg细胞比例和Foxp3 mRNA和蛋白表达(P<0.05)。结论 银甲片可有效地调节Th17/Treg平衡，进而调节Treg和Th17细胞介导的炎性反应，提高慢性输卵管模型大鼠受孕率。

Abstract:

OBJECTIVE To explore whether Yinjia tablets improve infertility in rats with chronic salpingitis by correcting the peripheral blood Th17/Treg balance. METHODS One hundred female Wistar rats, 82 of which were constructed as a rat model of chronic salpingitis. After successful modeling, the rats were randomly divided into model group, levofloxacin group (20 mg·kg^-1), low-dose Yinjia tablets group(0.38 g·kg^-1·d^-1), medium-dose Yinjia tablets group(0.76 g·kg^-1·d^-1) and high-dose Yinjia tablets group(1.52 g·kg^-1·d^-1), 16 rats in each group, and other 16 rats were set up as control group. The treatment groups were administered by gavage at different doses, and the model group and the control group were given the same amount of saline(20 mL·kg^-1·d^-1), once a day. The gavage was stopped for 1 d after 7 d, and 7 d was one cycle, with a total of 3 cycles of gavage. After administration, the rats were observed for conception and blood samples and oviductal tissues were collected. HE staining was used to observe the pathological changes of fallopian tube tissues; transmission electron microscopy was used to observe the ultrastructural changes of fallopian tube tissues; ELISA was used to detect TNF-α and IL-6 in serum; flow cytometry was used to detect the ratio of Treg and Th17 cells in peripheral blood; RT-qPCR and Western blotting were used to detect IL-17, IL-10, Foxp3, RORγt mRNA and protein expression. RESULTS Compared with the control group, the cilia and microvilli in the model group were sparse and the mitochondria were swollen; the TNF-α and IL-6 contents, the proportion of Th17 cells, and the expression of IL-17 and RORγt mRNA and protein in the oviduct tissues were significantly higher, and the fertility rate, IL-10, the proportion of Treg cells, and the expression of Foxp3 mRNA and protein in the rats were significantly lower(P<0.05). Compared with the model group, the high-dose group of Yinjia tablets improved the structure of oviduct of rats; significantly reduced the TNF-α, IL-6 content, Th17 cell proportion and IL-17, RORγt mRNA and protein expression in oviduct tissue of model rats; and significantly increased the conception rate and IL-10, Treg cell proportion and Foxp3 mRNA and protein expression of rats(P<0.05). CONCLUSION Yinjia tablets can effectively regulate Th17/Treg balance, which in turn regulates Treg and Th17 cell-mediated inflammatory responses and improves the conception rate in chronic salpingitis rats.