滋肾育胎丸对免疫性不孕模型大鼠卵母细胞生物学行为的调节作用

发布时间：2018-04-28 04:24

本文选题：滋肾育胎丸 + 免疫性不孕　；参考：《中国中西医结合杂志》2017年03期

【摘要】：目的探讨滋肾育胎丸对抗精子抗体(As Ab)阳性的不孕大鼠卵母细胞的干预作用。方法采用主动免疫建立As Ab阳性的免疫性不孕模型。将100只雌性SD大鼠随机分为空白对照组、模型组及滋肾育胎丸低、中、高剂量组(简称中药低、中、高剂量组),每组20只。造模22天开始灌胃给药,模型组给予5 m L/kg生理盐水灌胃,中药低、中、高剂量组分别给予0.075、0.100、0.150 g/m L滋肾育胎丸灌胃,每天1次,至36天。分别在造模21天及36天采用ELISA法测定As Ab和转化生长因子(transforming growth factor-β,TGF-β)水平。处死大鼠后取卵母细胞,采用实时定量聚合酶链反应(Real-time PCR)及Western blot分别检测TGF-βmRNA及其蛋白表达水平;采用流式细胞学(FCM)和末端终未标记(TUNEL)法测定卵母细胞凋亡率;采用细胞计数-8法(CCK-8)及5-乙炔基-2'脱氧尿嘧啶核苷(5-ethynyl-2'-deoxyuridine,Ed U)检测卵母细胞增殖水平;采用划痕实验及侵袭小室(Transwell)测定卵母细胞迁移与侵袭能力。结果与空白对照组比较,模型组卵母细胞凋亡增加,细胞增殖减少,迁移与侵袭能力减弱,TGF-βmRNA及其蛋白表达水平显著下调(P0.05);与模型组比较,中药高剂量组卵母细胞凋亡减少,细胞增殖增加,迁移与侵袭能力增强,TGF-βmRNA及其蛋白表达水平显著上调(均P0.05);且中药高剂量组作用明显优于中药低、中剂量组(P0.05)。结论应用高剂量滋肾育胎丸干预免疫性不孕大鼠可明显上调其卵母细胞TGF-β水平,改善卵母细胞功能。
[Abstract]:Objective to investigate the intervention effect of Zishen Yuet Wan (ZYT) on oocytes of infertile rats with positive sperm antibody (as Ab). Methods the immune infertility model with positive Asab was established by active immunity. 100 female Sprague-Dawley rats were randomly divided into blank control group, model group, low, medium and high dose groups (20 rats in each group). The model group was given intragastric administration of 5 m L/kg normal saline for 22 days, and the low, middle and high dose groups were given 0.075 ~ 0.100 ~ 0.150 g / ml Zishen Yu-fetus pill, once a day to 36 days. The levels of Asab and transforming growth factor- 尾 (TGF- 尾) were measured by ELISA method on the 21st and 36th day respectively. The expression of TGF- 尾 mRNA and its protein were detected by real-time quantitative polymerase chain reaction (PCR) and Western blot, and the apoptosis rate of oocytes was determined by flow cytology (FCM) and terminal unlabeled Tunel (Tunel). Cell count (CCK-8) and 5-ethynyl-2'deoxyuridine (Ed) were used to detect the proliferation of oocytes, and the migration and invasiveness of oocytes were measured by scratch test and invasive chamber transwelling. Results compared with the blank control group, the apoptosis of oocytes in the model group increased, the cell proliferation decreased, and the expression of TGF- 尾 mRNA and its protein decreased significantly in the model group, and the apoptosis of the oocytes in the high dose group decreased compared with that in the model group, and the expression of TGF- 尾 mRNA and its protein decreased in the model group. The expression level of TGF- 尾 mRNA and its protein was increased significantly (all P0.05%), and the effect of high dose group was better than that of low dose group, and the middle dose group was significantly higher than that of middle dose group (P0.05%). Conclusion the high dose Zishen Yuet pill can obviously upregulate the TGF- 尾 level of oocytes and improve the oocyte function in immunological infertility rats.
【作者单位】：南方医科大学附属花都医院围产医学中心;
【基金】：广东省科技厅科技项目(No.2015A030302004,No.2015A030302003) 广东省中医药局科技项目(No.20161191,No.20151061) 广东省自然科学基金资助项目(No.2016A030313419)
【分类号】：R285.5;R-332

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