滇桂艾纳香提取物促进DUB大鼠子宫收缩机制分析

Analysis of the mechanism of Blumea riparia extractpromoting uterine contractionvia in DUB rats

目的探讨滇桂艾纳香提取物通过促进子宫平滑肌收缩治疗功能失调性子宫出血(DUB)的作用机制。方法利用米非司酮联合米索前列醇建立DUB大鼠模型,成模的雌性大鼠40只,随机分5组:正常妊娠组、DUB模型组、益母草颗粒3.125 g·kg^(-1)组、滇桂艾纳香提取物0.4 g·kg^(-1)组以及0.2 g·kg^(-1)组,另取8只未受孕性成熟雌性大鼠作空白对照组,观察滇桂艾纳香提取物对DUB模型大鼠子宫平滑肌收缩的影响,Western blotting法测定大鼠子宫平滑肌中β_(2)肾上腺素受体(β_(2)AR)、环磷腺苷效应元件结合蛋白(CREB)及p-CREB蛋白表达水平,并检测大鼠子宫平滑肌中蛋白激酶A(PKA)活性和Ca^(2+)离子浓度。结果与DUB模型组相比,滇桂艾纳香提取物可提高大鼠子宫收缩频率(P<0.05),增加子宫平滑肌Ca^(2+)离子浓度,并且增加p-CREB/CREB蛋白表达(P<0.05);滇桂艾纳香提取物还可降低子宫平滑肌PKA活力(P<0.01),但对β_(2)受体表达没有明显影响。结论滇桂艾纳香提取物可能是通过抑制β_(2)受体的活化,间接增强转录调节因子CREB对与子宫平滑肌收缩相关的转录调控,从而促使子宫肌层收缩,产生治疗DUB的作用。

Objective To explore the therapeutic mechanism involved in uterine contraction promoting of Blumea riparia extract in dysfunctional uterine bleeding(DUB).Methods A total of 40 mature female rats were successfully modeled and randomly assigned into five groups:the normal pregnancy group,DUB model group,Motherwort Granules 3.125 g·kg^(-1) group,Blumea riparia extract 0.4 g·kg^(-1) group,and 0.2 g·kg^(-1) group.Additionally,eight non-pregnant mature female rats were selected as a blank control group.Mifepristone combined with Misocol were used to induce DUB in rats,and the effect of Blumea riparia extract on uterine contraction was evaluated.β_(2) adrenergic receptor,CREB and p-CREB proteins levels in the uterus smooth muscle were determined by Western blotting.Furthermore,PKA kinase activity and Ca^(2+)concentration in uterine smooth muscle were detected.Results Compared with the DUB model group,the uterine contraction frequency was increased by the treatment of Blumea riparia extract(P<0.05),and uterine Ca^(2+)concentration in smooth muscle as well as p-CREB/CREB protein expression were increased(P<0.05).Uterine PKA activity was reduced significantly after treating with Blumea riparia extract(P<0.01),while no significant changes in β_(2) receptor protein was observed.Conclusion Blumea riparia extract activated transcriptional regulation of CREB indirectly by inhibiting the activation of uterine β_(2) receptors.This may serve as one of the uterine contraction promoting mechanisms of Blumea riparia in DUB rats.