柴黄清胰活血颗粒对重症急性胰腺炎大鼠肾素-血管紧张素系统的影响

Effect of Chaihuang Qingyi Huoxue Granules on Renin-Angiotensin System in Rats with Severe Acute Pancreatitis

目的基于肾素-血管紧张素系统(RAS)探讨柴黄清胰活血颗粒(柴胡、生大黄、赤芍、白芍、厚朴等)对重症急性胰腺炎(SAP)大鼠的作用机制。方法将64只SD大鼠随机分为4组:假手术组、模型组、柴黄清胰活血颗粒组(4.42 g·kg^(-1))、卡托普利组(5 mg·kg^(-1)),各组再分为12、24 h两个亚组,每组8只。采用经胰胆管逆行注射3.5%牛磺胆酸钠复制SAP大鼠模型。卡托普利组腹腔注射给药;柴黄清胰活血颗粒组灌胃给药,每6 h灌胃1次。术后12、24 h采集标本,采用生化法检测血清淀粉酶(AMY)活性;HE染色法观察胰腺组织病理变化;化学发光法检测血清醛固酮(ALD)含量;ELISA法检测血清肾素(Renin)、血管紧张素转换酶(ACE)、血管紧张素Ⅱ(Ang-Ⅱ)含量;Western Blot法检测胰腺组织AT1R蛋白表达。结果在12、24 h同一亚组中,与假手术组比较,模型组大鼠的血清AMY活性均明显升高(P<0.05),胰腺组织病理评分明显升高(P<0.05),血清ALD、Renin、Ang-Ⅱ、ACE水平均明显上升(P<0.05),胰腺组织AT1R蛋白表达明显上调(P<0.05)。与模型组比较,柴黄清胰活血颗粒组、卡托普利组大鼠的血清AMY活性均明显下降(P<0.05),胰腺组织病理评分明显降低(P<0.05),血清ALD、Renin、Ang-Ⅱ、ACE水平均明显下降(P<0.05),胰腺组织AT1R蛋白表达明显下调(P<0.05)。与卡托普利组比较,柴黄清胰活血颗粒组大鼠的血清AMY活性均明显降低(P<0.05),胰腺组织病理评分明显降低(P<0.05),血清ALD、Renin、Ang-Ⅱ、ACE水平均明显下降(P<0.05)。结论柴黄清胰活血颗粒可能通过下调ACE-Ang-Ⅱ-AT1R经典轴的表达,抑制Renin、ALD的生成,从而发挥对SAP大鼠的保护作用。

Objective To investigate the mechanism of Chaihuang Qingyi Huoxue Granules(Bupleuri Radix,unprocessed Rhei Radix et Rhizoma,Paeoniae Radix Rubra,Paeoniae Radix Alba,Cortex Magnoliae Officinalis,etc.)on rats with severe acute pancreatitis(SAP)based on the renin-Ang-Ⅰotensin system(RAS).Methods Sixty-four SD rats were randomly divided into four groups:sham operation group,model group,Chaihuang Qingyi Huoxue Granules group(4.42 g·kg^(-1))and Captopril group(5 mg·kg^(-1)).Each group was further divided into 12-hour and 24-hour subgroups,with 8 rats in each group.SAP rat model was replicated by retrograde injection of 3.5%sodium taurocholate into the biliopancreatic duct.The Captopril group was intraperitoneally injected with Captopril(5 mg·kg^(-1)),and the Chaihuang Qingyi Huoxue Granules group was given intragastric administration,once every 6 hours.The serum amylase(AMY)activity was detected by biochemical method at 12 hours and 24 hours after operation.The pathological changes of pancreatic tissue were observed by HE staining.Serum aldosterone(ALD)content was detected by chemiluminescence.Serum Renin,angiotensin converting enzyme(ACE)and angiotensin II(Ang-II)were detected by ELISA.The expression of AT1R protein in pancreatic tissue was detected by Western Blot.Results In the same subgroup at 12 and 24 hours,compared with the sham operation group,the serum AMY activity of rats in the model group was significantly increased(P<0.05),the pathological score of pancreatic tissue was significantly increased(P<0.05),the levels of serum ALD,Renin,Ang-II and ACE were significantly increased(P<0.05),and the expression of AT1R protein in pancreatic tissue was significantly up-regulated(P<0.05).Compared with the model group,the serum AMY activity of rats in Chaihuang Qingyi Huoxue Granules group and Captopril group was significantly decreased(P<0.05),the pathological score of pancreatic tissue was significantly decreased(P<0.05),the levels of serum ALD,Renin,Ang-II and ACE were significantly decreased(P<0.05),and the expression of AT1R protein in pancreatic tissue was significantly down-regulated(P<0.05).Compared with the Captopril group,the serum AMY of the rats in the Chaihuang Qingyi Huoxue Granules group was significantly decreased(P<0.05),the pathological score of pancreatic tissue was significantly decreased(P<0.05),and the serum ALD,Renin,Ang-II and ACE levels were significantly decreased(P<0.05).Conclusion Chaihuang Qingyi Huoxue Granules may inhibit the production of Renin and ALD by down-regulating the expression of ACE-Ang-II-AT1R classical axis,thus exerting a protective effect on SAP rats.