加味龟鹿二仙汤对原发性卵巢功能不全小鼠卵巢miR-190表达的影响

Effects of modified Guilu Erxian Decoction on miR-190 expression of POI mice

目的:观察加味龟鹿二仙汤对原发性卵巢功能不全(POI)小鼠卵巢miR-190表达的影响,探讨加味龟鹿二仙汤对POI的干预作用及机制,为临床上中药治疗POI提供理论依据。方法:将连续两个动情周期正常的40只昆明雌性小鼠纳入实验,随机分为空白组、模型组、野山药复合营养软胶囊(DHEA)组、中药组。模型组、DHEA组及中药组连续15d腹腔注射环境化学物质(VCD)构建POI模型。其后中药组给予加味龟鹿二仙汤中药药液0.015mL/g灌胃,DHEA组给予DHEA药液0.015mL/g灌胃,空白组和模型组均给予0.9%氯化钠溶液0.015mL/g灌胃。4d为1个疗程,每4天停1d,共30d。观察小鼠动情周期和卵巢组织形态学变化;应用实时荧光定量PCR技术检测小鼠卵巢组织中miR-190表达水平。结果:与空白组比较,模型组小鼠卵巢miR-190表达明显升高(P<0.01);与模型组比较,DHEA组和中药组小鼠卵巢miR-190表达量均显著降低(P<0.01);且中药组小鼠卵巢miR-190表达低于DHEA组(P<0.01)。结论:加味龟鹿二仙汤能有效增加POI小鼠卵巢组织原始卵泡和初级卵泡数量,其机制可能与抑制卵巢mi R-190表达有关。

Objective: Investigated the effects of the modified Guilu Erxian Decoction on miR-190 expression of the POI mice. The intervention effect and mechanism of the modified Guilu Erxian Decoction on POI were researched, which can provide a theoretical basis for clinical treatment of POI. Methods: Forty female Kunming mice with continuous twice normal estrus cycles were involved in the experiments. They were randomly divided into control group, model group, DHEA group and traditional Chinese medicine(TCM) group. The mice of the model group, the DHEA group and the TCM group were intraperitoneal injection with VCD for 15 consecutive days to establish the mice model of POI. Then, the mice of the TCM group were given 0.015 mL/g modified Guilu Erxian Decoction to filled into the stomach, while the DHEA group were given 0.015 mL/g DHEA, and the control group and the model group were given 0.015 mL/g normal saline. The total treatment was 30 days, and 4 days were a period of treatment, stopping 1 day after every 4 days. Investigated the estrus cycles and morphology changes of ovarian tissue. Furthermore, the qRT-PCR were performed to detect the miR-190 expression in mice’s ovarian tissue. Results: The ovarian miR-190 expression of the model group was higher than that of the control group(P<0.01). The ovarian miR-190 expression of the DHEA group and the TCM group were lower than that of the model group(P<0.01), and the ovarian miR-190 expression of the TCM group was slightly lower than that of the DHEA group(P<0.01). Conclusion: The modified Guilu Erxian Decoction could effectively increase the primordial follicles and secondary follicles in POI mice’s ovarian tissue, the mechanism may be related to the inhibition of ovarian miR-190 expression.