马来酸依那普利叶酸片联合通心络对蛋氨酸诱导大鼠主动脉平滑肌细胞增殖的影响  

Effect of Enalapril Maleate and Folic Acid Tablets Combined with Tongxinluo on the Proliferation of Aortic Smooth Muscle Cells in Rats Induced by Methionine

作　　者：黄剑锋[1] 肖善花 黄绍烈[1] 段勇 胡永辉[3] 左汉恒[4] 巢胜吾[5] 陈可冀[6] 吴以岭 贾振华[8] 聂志华 袁丽霞 赵宇[11] 江小杰 HUANG Jian-feng;XIAO Shan-hua;HUANG Shao-lie;DUAN Yong;HU Yong-hui;ZUO Han-heng;CHAO Sheng-wu;CHEN Ke-ji;WU Yi-ling;JIA Zhen-hua;NIE Zhi-hua;YUAN Li-xia;ZHAO Yu;JIANG Xiao-jie(Deparment of UItrasonic Diagnosis,The First Affiliated Hospital of Nanchang University,Nanchang 330006;Department of Cardiovascular Medicine,The Third Hospital of Changsha,Changsha 410000;Department of Cardiovascular Medicine,Tai’an Hospital of Traditional Chinese Medicine,Shandong 271000;Department of Cardiovascular Medicine,Affiliated Hospital of Jining Medical University,Shandong 272000;Department of Cardiovascular Medicine,Shanghai People’s Liberation Army 455 Hospital,Shanghai 200052;National Clinical Research Center for Chinese Medicine Cardiology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091;Department of Cardiovascular Medicine,Hebei Institute of Integrated Traditional Chinese and Western Medicine,Shijiazhuang 050000;Department of Cardiovascular Medicine,Hebei Yiling Hospital,Shijiazhuang 050000;Department of Cardiovascular Medicine,Xiamen Haicang Hospital Department,Fujian 361000;Department of Cardiovascular Medicine,Zhengzhou Cardiovascular Hospital,Zhengzhou 450000;Department of Cardiovascular Medicine,The Fourth Affiliated Hospital of Nanchang University,Nanchang 330006;Department of Cardiovascular Medicine,Nanchang Third Hospital,Nanchang 330006)

机构地区：[1]南昌大学第一附属医院超声诊断科,南昌330006 [2]湖南长沙市第三医院心血管内科,长沙410000 [3]泰安市中医医院心血管内科,山东271000 [4]山东济宁医学院附属医院心血管内科,山东272000 [5]上海市解放军455医院心血管内科,上海200052 [6]国家中医心血管病临床医学研究中心,中国中医科学院西苑医院,北京100091 [7]河北省中西医结合医药研究院心血管内科,石家庄050000 [8]河北以岭医院心血管内科,石家庄050000 [9]厦门市海沧医院心血管内科,福建361000 [10]郑州市心血管病医院心血管内科,郑州450000 [11]南昌大学第四附属医院心血管内科,南昌330006 [12]南昌市第三医院心血管内科,南昌330006

出　　处：《中国中西医结合杂志》2022年第3期327-333,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：江西省科技厅重点项目(No.S2017YBYFE0196)。

摘　　要：目的探讨马来酸依那普利叶酸片联合通心络对蛋氨酸诱导的大鼠主动脉平滑肌细胞增殖的影响。方法通过蛋氨酸诱导大鼠主动脉平滑肌细胞增殖,采用马来酸依那普利叶酸片和通心络单独或者联合孵育细胞。实验随机分为对照组、蛋氨酸颗粒组、蛋氨酸颗粒加马来酸依那普利叶酸片组(简称EFA组)、蛋氨酸颗粒加通心络组(简称TXL组)及蛋氨酸颗粒加马来酸依那普利叶酸片加通心络组(简称EFA+TXL组);通过MTT实验探索蛋氨酸颗粒、马来酸依那普利叶酸片和通心络的最佳浓度。采用Western Blot和RT-qPCR检测葡萄糖调节蛋白94(GRP94)、半胱氨酸门冬氨酸特异性蛋白激酶12(Caspase-12)和葡萄糖调节蛋白78(GRP78)蛋白和mRNA表达,免疫荧光检测GRP94和Caspase-12表达。结果MTT结果显示:蛋氨酸颗粒最佳浓度为3.2 mmol/L,马来酸依那普利叶酸片和通心络最佳浓度分别选择10μg/mL和10μg/mL。与对照组比较,蛋氨酸颗粒组GRP94、Caspase-12和GRP78蛋白和m RNA表达明显升高(P<0.05);与蛋氨酸颗粒组比较,EFA组、TXL组、EFA+TXL组GRP94、Caspase-12和GRP78蛋白和m RNA表达明显下降(P<0.05);与EFA组和TXL组比较,EFA+TXL组中GRP94、Caspase-12和GRP78蛋白表达明显下降(P<0.05),GRP78 mRNA水平明显下降(P<0.05)。结论蛋氨酸可能通过内质网应激(ERS)途径参与并加重高血压引起的血管平滑肌重构的过程;马来酸依那普利叶酸片和通心络处理后能够通过抑制ERS保护血管平滑肌细胞,且联合作用效果更明显。Objective To explore the effect of enalaprilat maleate and folic acid tablets Combined with Tongxinluo on methione-induced proliferation of aortic smooth muscle cells in rats.Methods The proliferation of rat aortic smooth muscle cells was induced by the methionine and the cells were incubated with enalapril maleate and folate tablets and Tongxinluo alone or in combination.The cells were assigned to control group,methionine Granule group,methionine Granule+enalapril maleate and folic acid tablet group(EFA group),methionine Granule+Tongxinluo group(TXL group)and methionine Granule plus enalapril maleate and folic acid tablet+Tongxinluo group(EFA+TXL group).The optimum concentrations of methionine granules,enalapril maleate and folic acid tablets and Tongxinluo were determined by MTT assay.The protein and mRNA levels of glucose regulatory protein 94(GRP94),Caspase-12 and GRP78(GRP78)were measured by Western Blot and RT-qPCR.The expressions of GRP94 and Caspase-12 were detected by immunofluorescence assay.Results MTT results showed that the optimal concentration of methionine granules,enalapril maleate and folic acid tablets and Tongxinluo were 3.2 mmol/L,0 and 10μg/mL,respectively.Compared with the control group,the protein and mRNA expressions levels of GRP94,Caspase-12 and GRP78 were significantly increased in the methionine granule group(P<0.05).Compared with methionine granule group,the protein and mRNA expressions levels of GRP94,Caspase-12 and GRP78 were significantly decreased in the EFA group,TXL group and EFA+TXL group(P<0.05).Compared with the EFA group and the TXL group,the expression of GRP94,Caspase-12 and GRP78 protein in the EFA+TXL group decreased significantly(P<0.05).Conclusion Methionine may participate in and aggravate the process of vascular smooth muscle remodeling through endoplasmic reticulum stress(ERS)pathway;the treatment of enalapril maleate and folate Tablets or Tongxinluo could protect vascular smooth muscle cells by inhibiting ERS,and the combination of both is more promising.

关 键 词：马来酸依那普利叶酸片 通心络 同型半胱氨酸 主动脉平滑肌细胞 

分 类 号：R544.1[医药卫生—心血管疾病]