益气养阴活血通络方抑制JAK2/STAT3信号通路减轻糖尿病肾病大鼠肾组织细胞凋亡  

Yiqiyangyin Huoxuetongluo Prescription Reduces Renal Cell Apoptosis in Rats with Diabetic Nephropathy by Inhibiting JAK2/STAT3 Signaling Pathway

作　　者：杨凤文[1] 高飞 陈素枝[1] 袁国栋[1] 张倩[1] 檀金川 YANG Feng-wen;GAO Fei;CHEN Su-zhi;YUAN Guo-dong;ZHANG Qian;TAN Jin-chuan(Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China;Hebei University of Chinese Medicine,Shijiazhuang 050200,China)

机构地区：[1]河北省中医院,石家庄050011 [2]河北中医学院,石家庄050200

出　　处：《中国实验方剂学杂志》2021年第11期89-97,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省中医药管理局项目(2015013,2016025)。

摘　　要：目的:观察益气养阴活血通络方对糖尿病肾病(DN)大鼠Janus酪氨酸蛋白激酶2(JAK2)/信号传导及转录激活因子3(STAT3)信号通路及细胞凋亡的影响,探讨其干预DN的作用机制。方法:SD大鼠100只随机分为造模组80只,正常组20只,高糖高脂饮食联合一次性腹腔注射链脲佐菌素(STZ)建立糖尿病肾病大鼠模型。造模结束后每组随机处死3只大鼠,光镜和电镜下观察肾脏组织病理学变化确认造模成功。将造模成功的大鼠随机分为模型组(生理盐水,等体积)、益气养阴活血通络方低中高剂量组(益气养阴活血通络方,5.775,11.550,23.100 g·kg^(-1))和厄贝沙坦组(厄贝沙坦片,0.016 g·kg^(-1)),各组分别给予相应剂量药物灌胃,每日1次,连续干预16周。给药结束后检测大鼠24 h尿蛋白(UTP)水平,血清总胆固醇(TC),甘油三酯(TG),肌酐(SCr),尿素氮(BUN)水平;蛋白免疫印迹法(Western blot)检测JAK2/STAT3信号通路关键蛋白的表达水平;免疫组化法(IHC)检测大鼠肾组织中B细胞淋巴瘤-2相关X蛋白(Bax),B细胞淋巴瘤-2(Bcl-2),紧密连接相关蛋白-1(ZO-1),肌动蛋白4(actinin-4)蛋白表达水平。结果:与正常组比较,模型组大鼠UTP,血清中TC,TG,BUN,SCr水平均有明显升高(P<0.05);大鼠肾组织病理损伤严重;磷酸化JAK2(p-JAK2),磷酸化STAT3(p-STAT3),Bax蛋白表达水平明显上调;肾组织细胞凋亡明显增多;Bcl-2,ZO-1,actinin-4蛋白表达水平下调(P<0.05)。与模型组比较,益气养阴活血通络方和厄贝沙坦组大鼠UTP,血清中TC,TG,BUN,SCr水平均有不同程度的下降(P<0.05);大鼠肾组织病理损伤有所减轻;p-JAK2,p-STAT3,Bax蛋白表达水平不同程度的下降;肾组织细胞凋亡减轻;Bcl-2,ZO-1,actinin-4蛋白表达水平不同程度的升高(P<0.05)。结论:益气养阴活血通络方药可通过抑制JAK2/STAT3信号通路的激活,减轻肾脏细胞凋亡,改善DN的预后。Objective:To observe the effect of Yiqiyangyin Huoxuetongluo prescription on janus kinase2/signal transducer and activator of transcription 3(JAK2/STAT3)signaling pathway and cell apoptosis in rats with diabetic nephropathy(DN),and to explore the mechanism of its intervention in DN. Method:A total of 100 SD rats were randomly divided into an experimental group(n=80)and a normal group(n=20). The DN model was induced by high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin(STZ)in the experimental group,and confirmed by the pathological changes of kidney tissues in rats(three in each group)observed under light and electron microscopes. The model rats were randomly divided into a model group(normal saline,equal volume),low-,medium-,and high-dose(5.775,11.550,and 23.100 g·kg^(-1))Yiqiyangyin Huoxuetongluo prescription groups,and an irbesartan group(irbesartan tablets,0.016 g·kg^(-1)).After drug intervention(i. g.,once a day for 16 consecutive weeks),the 24-hour urine total protein(UTP),serum total cholesterol(TC),triglyceride(TG),creatinine(SCr),and blood urea nitrogen(BUN)levels of the rats were measured. Western blot was used to detect the protein expression of JAK2/STAT3 signaling pathway. Immunohistochemistry was used to determine the protein expression of B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),zonula occludens-1(ZO-1),and actinin-4 in rat kidney tissues. Result:Compared with the normal group,the model group exhibited elevated UTP,serum TC,TG,BUN,and SCr levels(P<0.05),severe pathological damage of rat kidney tissues,up-regulated expression of phospho-JAK2(p-JAK2),phospho-STAT3(p-STAT3),and Bax,increased renal cell apoptosis,and diminished expression of Bcl-2, ZO-1, and actinin-4(P<0.05). Compared with the model group, the Yiqiyangyin Huoxuetongluo prescription group and the irbesartan group showed dwindled UTP,serum TC,TG,BUN,and SCr levels(P<0.05), relieved pathological damage of rat kidney tissues, down-regulated p-JAK2, p-STAT3, and Bax expression,and up-reg

关 键 词：益气养阴活血通络方 糖尿病肾病(DN) Janus酪氨酸蛋白激酶2(JAK2)/信号传导及转录激活因子3信号通路(STAT3) 细胞凋亡 

分 类 号：R2-0[医药卫生—中医学] R22