[滋肾育泰丸对免疫不孕症模型大鼠卵母细胞生物学行为的调节作用]。

中国中西医结合杂志 Pub Date : 2017-03-01
Xiao-Li Zhang, Xiao-Yong Zhao
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引用次数: 0

摘要

目的观察滋肾育泰丸(ZYP)对抗精子抗体(AsAb)阳性不育大鼠oo细胞的干预作用。方法采用主动免疫法建立AsAb阳性免疫不育模型。将100只雌性SD大鼠随机分为空白对照组、模型组和ZYP低、中、高剂量组,每组20只。从造模第22天至第36天,各组大鼠灌胃给予不同药物,每天1次。模型组大鼠灌胃给予生理盐水5 mL/kg。ZYP为0。ZYP低、中、高剂量组大鼠分别给予075、0.100、0.150 g/mL。ELISA法检测血清AsAb和TGF-β水平。大鼠处死后收集卵母细胞。采用实时定量聚合酶链反应(RT-PCR)和Western blot检测mRNA和蛋白表达水平。采用流式细胞术(FCM)和转移酶介导的三磷酸脱氧尿苷-生物素缺口末端标记法(TUNEL)检测细胞凋亡率。采用细胞计数试剂盒8 (CCK-8)和5-乙基-2′-脱氧尿苷(EdU)检测卵母细胞增殖水平。采用入侵室(Transwell)和划痕实验测定了虫卵的迁移和入侵。结果与空白对照组相比,大鼠卵母细胞凋亡增加,增殖能力减弱,迁移和侵袭能力增强,TGF-β mRNA和蛋白表达水平显著下调(P
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Regulatory Effect of Zishen Yutai Pill on the Biological Behavior of Oocytes in Immune Infertility Model Rats].

Objective To observe the intervention effect of Zishen Yutai Pill (ZYP) on the oo- cytes of infertile rats with positive anti-sperm antibody (AsAb). Methods AsAb positive immune infertil- ity model was established using active immunity. Totally 100 female SD rats were randomly divided into the blank control group, the model group, low, middle, and high dose ZYP groups, 20 in each group. Dif- ferent medications were administered to rats in corresponding groups by gastrogavage from the 22nd day of modeling to the 36th day, once per day. Normal saline (5 mL/kg) was administered to rats in the model group by gastrogavage. ZYP at 0. 075, 0.100, 0.150 g/mL was respectively administered to rats in low, middle, and high dose ZYP groups. AsAb and transforming growth factor β (TGF-β) levels were meas- ured by ELISA. Oocytes were collected after rats were sacrificed. mRNA and protein expression levels were detected using Real-time quantitative polymerase chain reaction ( RT-PCR) and Western blot. Oo- cyte apoptosis rate was determined by flow cytometry (FCM) and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL). The oocyte proliferation level was detected by cell count- ing kit 8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU). Oodyte migration and invasion was determined u- sing invasion chamber (Transwell) and scratch experiment. Results Compared with the blank control group, the apoptosis of oocytes increased, the proliferation of oocytes was reduced, the capabilities of migration and invasion were enhanced, mRNA and protein expression levels of TGF-β were significantly down-regulated (P <0. 05). Compared with the model group, the apoptosis of oocytes was reduced, the proliferation of oocytes was increased, the capabilities of migration and invasion were enhanced, mRNA and protein expression levels of TGF-β were significantly up-regulated in the high dose ZYP group (P < 0. 05). Besides, their effects were obviously superior in the high dose ZYP group than in low and middle dose ZYP groups (P <0. 05). Conclusion The application of high dose ZYP could obviously up-regulate the level of TGF-β and improve the function of oocytes.

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