基于血管生成探讨补肾养精颗粒对早发性卵巢功能不全大鼠模型的影响

Study of the Effects of Bushen Yangjing Granules on a Rat Model of Early Ovarian Insufficiency Based on Angiogenesis

目的基于血管生成及凋亡相关因子探讨补肾养精颗粒治疗早发性功能不全的机制。方法选定动情周期正常的60只SD雌性大鼠,随机分为空白组、模型组、补肾养精颗粒组(低剂量组(1.75 g·kg^(-1))、中剂量组(3.50 g·kg^(-1))、高剂量组(7.00 g·kg^(-1))、补佳乐组。通过阴道涂片观察动情周期的变化;ELISA检测大鼠血清E2、AMH、NF-κB、MMP-2、Caspase-3、Caspase-9浓度;Western blot检测卵巢组织中MMP-2、MMP-9、VEGF、Caspase-3的表达;HE染色观察大鼠卵泡发育及血管分布情况。结果(1)模型组大鼠的动情周期明显延长,予补肾养精颗粒干预后明显缩短;(2)模型组的血清E2、AMH、NF-κB、MMP-2浓度较低,予补肾养精颗粒干预后其表达升高(P<0.05);(3)模型组的Caspase-3、Caspase-9表达偏高,予补肾养精颗粒干预后其表达降低(P<0.05);(4)在卵巢组织MMP-2、MMP-9、VEGF的蛋白含量方面,模型组均低于空白组,予补肾养精颗粒干预后其表达升高;(5)在HE方面,模型组的卵泡数目及血管较少,予补肾养精颗粒干预后可改善。结论补肾养精颗粒可通过升高血管生成相关因子NF-κB、MMP-2、MMP-9、VEGF的表达,降低凋亡相关因子Caspase-3、Caspase-9的表达,进而改善血清E2、AMH水平,促进卵巢血管的形成及卵泡的发育。

Objective To study the mechanism of early-onset sexual insufficiency based on angiogenesis and apoptosis-related factors treated by Bushen Yangjing Granule.Methods A total of 60 SD female rats with normal estrus cycle were randomly divided into blank group,model group,Bushen Yangjing Granule group:low dose group(1.75 g·kg^(-1)),medium dose group(3.50 g·kg^(-1)),high dose group(7.00 g·kg^(-1))and Progynova group.We observed the change of estrus cycle by vaginal smear.Concentration of serum E2,AMH,NF-κB,MMP-2,Caspase-3,Caspase-9 were detected by ELISA.The expression of MMP-2,MMP-9,VEGF,Caspase-3 were detected by Western blot.The follicular development and blood vessel distribution in rats were observed by HE.Results(1)In terms of estrous cycle,the model group was significantly prolonged.After the intervention of Bushen Yangjing Granule,it was shortened obviously;(2)The concentrations of E2,AMH,NF-κB and MMP-2 were relatively low in the model group,which were improved after the intervention of Bushen Yangjing granules(P<0.05);(3)The expression of Caspase-3 and caspase-9 in the model group was relatively high,which could be significantly reduced after the intervention of Bushen Yangjing Granules.(4)The protein content of MMP-2,MMP-9 and VEGF in the ovarian tissue of the model group was relatively low,which could be significantly improved after the intervention of Bushen Yangjing Granules.(5)The number of follicles and blood vessels in the model group were relatively less,which could be improved by the intervention of Bushen Yangjing Granules and Progynova.Conclusion Bushen Yangjing Granule can increase the expression of angiogenesis related factors(NF-κB,MMP-2,MMP-9 and VEGF)reduce the expression of apoptosis related factors(caspase-3 and caspase-9),improve the level of serum E2 and AMH,promote the formation of ovarian blood vessels and the development of follicles.