养精种玉汤经FTO调控卵巢细胞自噬的机制研究

Study on the Mechanism of Yang Jing ZhongYu Decoction Regulating Ovarian Autophagy by FTO

目的研究养精种玉汤经FTO对卵巢储备功能减退(DOR)模型大鼠细胞自噬的机制。方法正常SD雌性大鼠,雷公藤甲素制备DOR大鼠模型,设空白组、模型组、DHEA组、养精种玉汤低、中、高剂量组6组,每组各10只。ELISA检测血清E_(2)、FSH、AMH浓度;Western Blotting测定卵巢组织中FTO、ULK1、LC3BⅡ的蛋白表达量。结果(1)与空白组比较,模型组大鼠E_(2)、AMH下降(P<0.05)而FSH上升(P<0.05),FTO、ULK1、LC3BⅡ表达水平均下调(P<0.05);(2)与模型组比较,中药高剂量组E_(2)、FSH显著升高(P<0.05),中药中剂量与低剂量组AMH显著升高(P<0.05),中药高剂量组和中剂量组FTO、ULK1、LC3B-Ⅱ蛋白相对表达显著升高(P<0.05),低剂量组及DHEA组ULK1蛋白表达显著升高(P<0.05)。结论养精种玉汤能通过上调RNA-m6A甲基化修饰的脱甲基酶FTO的表达,启动卵巢细胞自噬,从而调节卵巢性激素水平,改善DOR模型大鼠的卵巢功能。

OBJECTIVE To study the effect of“YangJing ZhongYu Decoction”( YJZYD) on the expression of FTO in ovary of rats with diminished ovarian reserve( DOR) function and the mechanism of regulating ovarian autophagy.METHODS SD female rats with normal estrous cycle were selected,and triptolide was used to make a model.A blank group,a model group,a DHEA group,and low,medium-and high-dose groups of YJZYD were set up.10rats in each group.Ovarian histological examination was performed by HE staining;serum E_(2),FSH,AMH concentrations were detected by ELISA;the remative protein expression levels of FTO,ULK1 and LC3B-Ⅱ in ovarian tissue were determined by Western Blotting.RESULTS(1) Compare with control group,E_(2) and AMH decreased( P <0.05),while FSH increased( P < 0.05),and the expression levels of FTO,ULK1 and LC3B-Ⅱ decreased( P <0.05) in model group;(2)Compared with the model group,E_(2)and FSH increased significantly in the high-dose group( P < 0.05),AMH increased significantly in the medium and low-dose groups( P < 0.05),the relative protein expression of FTO,ULK1 and LC3B-Ⅱ increased significantly in the high-dose group and medium-dose group( P <0.05),and the expression of ULK1 protein increased significantly in the low-dose group and DHEA group( P <0.05).CONCLUSION YJZYD can upregulate the expression of RNA-m6A methylation modified demethylase FTO and initiate ovarian autophagy,so as to regulate the level of ovarian sex hormone and improve the ovarian function of DOR model rats.