针药合用通过调控TGF-β1信号通路促进肾虚型排卵障碍大鼠卵巢颗粒细胞形态的恢复

Combination of acupuncture and traditional Chinese medication promotes the morphological recovery of ovarian granulosa cells by regulating TGF-β1 signaling pathway

目的探索针药合用促进肾虚型排卵障碍大鼠卵巢颗粒细胞形态恢复的具体机制。方法利用羟基脲灌胃方法构建肾虚型排卵障碍大鼠动物模型,对模型组大鼠分别给予针刺(取穴气海、关元、双侧三阴交)+中药(补肾健脾方水煎剂)(针药合用组)、中药+假针(中药组)、针刺+安慰剂中药(针刺组)、假针+安慰剂中药(空白对照组)处理。取各组大鼠新鲜的卵巢组织,利用实时荧光定量PCR和Western blot技术分别检测其余各组大鼠卵巢新鲜组织中的转化生长因子-β1(TGF-β1)、smad3以及smad7的mRNA和蛋白表达情况。结果3个实验组的TGF-β1、smad3的mRNA和蛋白水平均较空白对照组明显降低,而smad7的mRNA和蛋白水平均较空白对照组明显升高。同时,针药合用组的TGF-β1、smad3的mRNA和蛋白水平较中药组、针刺组及空白对照组均明显降低,而smad7的mRNA和蛋白水平较中药组、针刺组及空白对照组均明显升高。结论针药合用通过抑制TGF-β1信号通路的活化进而促进肾虚型排卵障碍大鼠卵巢颗粒细胞形态的恢复。

Objective To investigate the mechanism of promoting the morphological recovery of ovarian granulosa cells in kidney-deficiency ovulatory disorder rats by combination of acupuncture and traditional Chinese medication.Methods The animal model of ovulation obstacle of kidney deficiency type was established by intragastric hydroxyurea lavage.Then the rats of model group were treated with acupuncture(Qi-sea,Guangyuan and bilateral Sanyinjiao)plus traditional Chinese medication(decoction for tonifying kidney and strengthening spleen)(Experimental Group 1);traditional Chinese med medication plus false needle(Experimental Group 2);acupuncture plus placebo(Experimental Group 3);and false needle plus placebo(Negative Control Group).The expression of TGF-1,smad3 and smad7 mRNA and protein in fresh ovary tissues of each group were detected by real-time PCR and Western blot,respectively.Results The mRNA and protein levels of TGF-β1 and smad3 in the three experimental groups were significantly lower than those in the control group,while the mRNA and protein levels of smad7 were significantly higher than those in the control group.Meanwhile,TGF-β1 and smad3 mRNA and protein levels in experimental Group 1 were significantly lower than those in experimental Group 2,3 and control group,while the mRNA and protein levels of smad7 in experimental Group 1 were significantly higher than those in experimental Group 2,3 and control Group.Conclusion The combination of acupuncture and traditional Chinese medication can inhibit the activation of TGF-β1 signaling pathway and promote the morphological recovery of ovarian granulosa cells in kidney-deficiency ovulatory rats.