Zhang Xiao-Li, Zhao Xiao-Yong
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2017 Mar;37(3):351-355.
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.
目的 观察滋肾育胎丸(ZYP)对抗精子抗体(AsAb)阳性不育大鼠卵母细胞的干预作用。方法 采用主动免疫法建立AsAb阳性免疫性不育模型。将100只雌性SD大鼠随机分为空白对照组、模型组、ZYP低、中、高剂量组,每组20只。从造模第22天至第36天,对相应组大鼠进行灌胃给予不同药物,每天1次。模型组大鼠灌胃给予生理盐水(5 mL/kg)。ZYP低、中、高剂量组大鼠分别灌胃给予0.075、0.100、0.150 g/mL的ZYP。采用酶联免疫吸附测定(ELISA)法检测AsAb和转化生长因子β(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)和蛋白质免疫印迹法检测mRNA和蛋白质表达水平。采用流式细胞术(FCM)和脱氧尿苷三磷酸生物素缺口末端标记法(TUNEL)检测卵母细胞凋亡率。采用细胞计数试剂盒8(CCK-8)和5-乙炔基-2'-脱氧尿苷(EdU)检测卵母细胞增殖水平。采用侵袭小室(Transwell)和划痕实验检测卵母细胞迁移和侵袭能力。结果 与空白对照组比较,卵母细胞凋亡增加,卵母细胞增殖减少,迁移和侵袭能力增强,TGF-β的mRNA和蛋白质表达水平显著下调(P<0.05)。与模型组比较,ZYP高剂量组卵母细胞凋亡减少,卵母细胞增殖增加,迁移和侵袭能力增强,TGF-β的mRNA和蛋白质表达水平显著上调(P<0.05)。此外,ZYP高剂量组的作用明显优于低、中剂量组(P<0.05)。结论 高剂量ZYP的应用可明显上调TGF-β水平,改善卵母细胞功能。