利拉鲁肽经AMPK途径对平滑肌细胞KCa3.1蛋白表达的影响  

The Influence of Liraglutide on KCa3.1 of Smooth Muscle Cells via AMPK Pathway

作　　者：董鹏[1] 李小红[1] 王俊红[1] 刘超峰 王玉环[1] 张春虹 DONG Peng;LI Xiaohong;WANG Junhong;LIU Chaofeng;WANG Yuhuan;ZHANG Chunhong(The Second Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710054,Shaanxi,China)

机构地区：[1]西安交通大学第二附属医院,西安710054 [2]陕西省中医院

出　　处：《中西医结合心脑血管病杂志》2020年第4期575-578,共4页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：陕西省社会发展科技攻关项目(No.2016SF-106)。

摘　　要：目的探讨利拉鲁肽经激活腺苷酸活化的蛋白激酶(AMPK)途径对平滑肌细胞KCa3.1蛋白表达的影响。方法制备SD大鼠血管平滑肌细胞模型,大鼠主动脉平滑肌细胞采用利拉鲁肽进行干预,测定未加入利拉鲁肽及加入利拉鲁肽15 min后磷酸化AMPK(PAMPK)/AMPK比值。将平滑肌细胞分为4组培养:A组为正常组,B组采用血管紧张素Ⅱ100 nmol/L,C组采用利拉鲁肽1μmol/L+血管紧张素Ⅱ100 nmol/L,D组采用利拉鲁肽1μmol/L+AMPK阻滞剂4μmol/L+血管紧张素Ⅱ100 nmol/L。培养72 h后使用Western-Blotting检测KCa3.1蛋白表达。结果显微镜下可见细胞呈长梭状、三角状,α-actin细胞免疫荧光染色后,胞浆着色为红色,确认为血管平滑肌细胞。利拉鲁肽与大鼠血管平滑肌作用15 min后,与基线比较PAMPK水平明显上升,PAMPK与AMPK比值明显上调,差异有统计学意义(P<0.05)。C组平滑肌细胞KCa3.1蛋白表达较B组下降(P<0.05)。结论利拉鲁肽可激活AMPK细胞通路,抑制平滑肌细胞上KCa3.1蛋白表达。Objective To observe the influence of liraglutide on KCa3.1 of smooth muscle cells via adenosine 5′-monophosphate-activated protein kinase(AMPK)pathway.Methods The aortic smooth muscle cells of sprague-dawley(SD)rats were treated with liraglutide,and phosphorylated AMPK(PAMPK)/AMPK ratio was measured at 0,and 15 minute after adding liraglutide.The smooth muscle cells were divided into 4 groups:A group was normal group,B group was treated with angiotensinⅡ(AngⅡ)100 nmol/L,C group was treated with liraglutide 1μmol/L+AngⅡ100 nmol/L,D group was treated with liraglutide 1μmol/L+AMPK blocker 4μmol/L+AngⅡ100 nmol/L.The expression of KCa3.1 of vascular smooth muscle cells were detected using Western-Blotting after 72 h.Results The cells were long fusiform and triangular under the microscope.After immunofluorescence staining ofα-actin cells,the cytoplasm was red,which was confirmed as vascular smooth muscle cells.Compared with the baseline value,the levels of PAMPK and PAMPK/AMPK were significantly increased after 15 minutes of liraglutide treatment(P<0.05).The expression of KCa3.1 in C group was lower than that in B group(P<0.05).Conclusion Liraglutide could activate AMPK cell pathway,and inhibit the expression of KCa3.1 ion channel in smooth muscle cells.

关 键 词：平滑肌细胞 利拉鲁肽 腺苷酸活化的蛋白激酶 KCa3.1蛋白 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]