加味黄芪赤风汤含药血清对LPS诱导的小鼠肾小球系膜细胞ColⅣ、MMP-2及TIMP-2表达的影响  

Effect of Modified Hangqi Chifeng Decoction Containing Serum on the Expression of Col Ⅳ, MMP-2, and TIMP-2 in Glomerular Mesangial Cells Induced by LPS

作　　者：刘红霞[1,2] 张昱[1,2] 李澎[3] 高艳红[1,4] 李双[1,4] 于子凯 

机构地区：[1]中国中医科学院西苑医院肾病科,北京100091 [2]中国中医科学院研究生院,北京100091 [3]中国中医科学院西苑医院实验研究中心,北京100091 [4]北京中医药大学研究生院,北京100029

出　　处：《中国中西医结合杂志》2016年第5期592-596,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：中国中医科学院基本科研业务费自主选题资助项目(No.ZZ0708105)

摘　　要：目的探讨加味黄芪赤风汤对脂多糖(LPS)诱导的小鼠肾小球系膜细胞(glomerular mesangial cells,GMCs)细胞外基质(extracellular matrix,ECM)Ⅳ型胶原蛋白(collagenⅣ,ColⅣ)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)及基质金属蛋白酶抑制酶2(tissue inhibitor of metalloproteinase-2,TIMP-2)水平的影响。方法采用血清药理学方法制备正常血清及替米沙坦、加味黄芪赤风汤高、中、低剂量含药血清。体外培养GMCs,以LPS作为刺激因子,诱导系膜细胞增殖。将GMCs分为正常组、模型组、替米沙坦组及加味黄芪赤风汤高、中、低剂量组(简称中药高、中、低剂量组)。采用ELISA法检测各组GMCs上清液中ColⅣ含量,Western blot法检测GMCs中MMP-2及TIMP-2蛋白表达水平。结果与正常组比较,LPS刺激72 h后,模型组GMCs上清ColⅣ含量明显增多,系膜细胞MMP-2及TIMP-2蛋白表达均明显上调,MMP-2/TIMP-2比例下调(均P<0.01);与模型组比较,中药高、中剂量组及替米沙坦组ColⅣ含量明显减少(P<0.01);中药中、低剂量组MMP-2蛋白表达明显下调(P<0.01,P<0.05);替米沙坦组及中药高、中、低剂量组TIMP-2蛋白表达均下调(P<0.01)。中药高剂量组TIMP-2蛋白表达水平明显低于中药低剂量组(P<0.01);替米沙坦组、中药高、中剂量组MMP-2/TIMP-2比例明显上调(P<0.01)。结论加味黄芪赤风汤可调节LPS诱导的GMSc ECM中MMP-2/TIMP-2比例失调,抑制ECM中ColⅣ的过度产生,促进ECM降解,减少ECM积聚,从而延缓肾小球硬化的进程。Objective To explore the effect of Modified Hangqi Chifeng Decoction（MHCD） on levels of collagen type Ⅳ（ColⅣ）, matrix metalloproteinase-2（MMP-2）, tissue inhibitor of metalloproteinase-2（TIMP-2） in extracellular matrix（ECM） of glomerular mesangial cells（GMCs） in LPS induced mice. Methods Normal serum and telmisartan, high, medium, low dose MHCD containing serums were prepared by using serum pharmacology meth-od. GMCs were cultured in vitro. The proliferation of mesangial cells were induced using LPS as stimulating factor.GMCs were divided into six groups, i.e., the normal group, the model group, the telmisartan group, high, medium and low dose MHCD groups. ColⅣ content in the supernatant of mesangial cells was detected using ELISA. Protein expressions of MMP-2 and TIMP-2 were detected using Western blot. Results Compared with the normalgroup, ColⅣ content obviously increased in the model group after 72-h LPS stimulation;protein expressions of MMP-2 and TIMP-2 were obviously up-regulated, and MMP-2/TIMP-2 ratio was down-regulated in the model group（P〈0. 01）. Compared with the model group, ColⅣ content obviously decreased in high and medium dose MHCD groups and the telmisartan group（P〈0. 01）;protein expressions of MMP-2 were obviously down-regulated in medium and low dose MHCD groups（P〈0. 01, P〈0. 05）;the protein expression of TIMP-2 was obviously down-regulated in high, medium, low dose MHCD groups and the telmisartan group（P〈0. 01）. The protein expression of TIMP-2 was obviously lower in the high dose MHCD group than in the low dose MHCD group（P〈0. 01）. MMP-2/TIMP-2 ratio was obviously up-regulated in the telmisartan group, high and medium dose MHCD groups（P〈0. 01）. Conclusion MHCD could regulate disordered MMP-2/TIMP-2 ratio in LPS induced ECM, inhibit excessive production of ColⅣ in ECM, promote the degradation of ECM, reduce the accumulation of ECM, thereby, delaying the process of glomerular sclerosis.

关 键 词：加味黄芪赤风汤 肾小球系膜细胞 Ⅳ型胶原蛋白 基质金属蛋白酶2 基质金属蛋白酶抑制酶2 肾小球硬化 

分 类 号：R285.5[医药卫生—中药学]