益智仁盐制前后组成缩泉丸对下丘脑-垂体-肾上腺皮质系统与β_3-AR、M_3R表达的比较

Effects of Fructus Alpiniae Oxyphyllae after salt processing in Suoquan wan on HPA and expression of β_3-AR and M_3R

目的:观察益智仁盐制前后组成缩泉丸对下丘脑-垂体-肾上腺皮质系统(HPA)的调节与膀胱β肾上腺素能受体(β_3-AR)与胆碱能神经受体(M_3R)mRNA和蛋白表达的影响,拟阐明盐制入肾-肾主水的机制。方法:采用酶联免疫吸咐法(ELISA)测定血浆中促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ATCH)和cAMP的浓度,RT-PCR测定β_3-AR与M_3R mRNA的表达,Western blot测定β_3-AR与M_3R的蛋白表达。结果:与模型组比较,缩泉丸中益智仁盐制后能提高肾阳虚多尿大鼠血浆中CRH、ATCH和cAMP的浓度(P<0.05),增加β_3-AR mRNA、蛋白表达(P<0.05),降低M_3RmRNA、蛋白表达(P<0.05),且比生品组作用增强(P<0.05)。结论:缩泉丸中益智仁盐制后对腺嘌呤所致肾阳虚多尿模型大鼠HPA轴有上调作用而调节体液平衡,同时也通过调节膀胱逼尿肌β_3-AR、M_3R mRNA和蛋白表达增强缩尿作用,证实了缩泉丸中用盐益智仁的科学性。

OBJECTIVE To study on the influence of Fructus Alpiniae Oxyphyllae（FAO）after salt processing in Suoquan wan（SQW）to HPA and expression of M3 R and β3-AR,and investigate the mechanism of salt processing to enter and regulate the kidney.METHODS The contents of CRH,ATCH and cAMP were determined by ELISA.RT-PCR and Western blot were chosen to detect the expression of M3 R and β3-AR mRNA and proteins.RESULTS Compared with the model group,salt-processed SQW increased the concentrations of CRH,ATCH and cAMP（P0.05）,and the expression of β3-AR mRNA and protein（P0.05）,decreased the expression of M3 R mRNA and protein（P0.05）.After salt processing,FAO effects were strengthened obviously compared with the raw drug（P0.05）.CONCLUSION Salt-processed SQW can regulate the HPA in adenine induced kidney yang deficiency with polyuria,and adjust fluid balance.Polyuria arresting effects can be enhanced by adjusting effects of M3 R and β3-AR mRNA and protein expression in detrusor of bladder,which provides the scientific evidence for using salt processed FAO in SQW.