丹参单体IH764-3对马兜铃酸致人肾小管上皮细胞损害的保护机制研究  

The protective effect and action mechanism of Salvia miltiorrhiza monomer-IH764-3 on aristolochic acid inducedrenal tubular epithelial cell injury

作　　者：胡波[1] 范红伟[2] 王燕午[3] 马广骏[4] 

机构地区：[1]中国人民解放军白求恩医务士官学校内科教研室,石家庄市050081 [2]河北省石家庄市第四医院 [3]中国人民解放军白求恩医务士官学校门诊部,石家庄市050081 [4]中国人民解放军第260医院医教处

出　　处：《河北医药》2016年第7期982-986,共5页Hebei Medical Journal

基　　金：全军青年培育项目(编号:13QNP187)

摘　　要：目的探讨丹参单体IH764-3在马兜铃酸(aristolochic acid,AA)导致人肾小管上皮细胞系(HK-C)损害中的保护作用及机制。方法以含体积分数0.10胎牛血清的DMEM培养液培养HK-C细胞72 h。实验分为3大组(8小组),AA组在培养液中加入浓度为20 mg/L的AA;观察组在培养液中加入浓度为20 mg/L的AA和不同浓度丹参单体IH764-3;对照组培养液中不加AA。培养24、48、72 h后,酶联免疫吸附试验(ELISA)检测各细胞培养液中转化生长因子β1(TGF-β1)、纤溶酶原激活物抑制剂1(PAI-1)、基质金属蛋白酶抑制剂1(TIMP-1)、金属蛋白酶2(MMP-2);采用MTT法观察HK-C细胞活性,实时荧光定量PCR技术检测凋亡蛋白Caspase-3mRNA的表达情况。结果 AA刺激后,诱导TGF-β1、TIMP-1、PAI-1表达增加,MMP-2表达减少(P<0.05)。经丹参单体IH764-3干预后,TGF-β1、TIMP-1、PAI-1表达减少,MMP-2表达增加,其中以IH764-3 80 mg/L效果最佳(P<0.05)。AA组及观察组较对照组HK-C细胞的活性均受到抑制,细胞的凋亡率增加,Caspase-3mRNA表达增高(P<0.05)。观察组较AA组细胞活性明显增高,细胞的凋亡率明显下降,Caspase-3mRNA表达降低,观察组中以丹参单体IH764-3 80 mg/L效果最佳(P<0.05)。结论丹参单体IH764-3可明显提高HK-C细胞抵抗AA导致细胞损伤的能力,保护HK-C细胞,其具体途径可能与抑制TGF-β1、TIMP-1、PAI-1、Caspase-3mRNA,升高MMP-2的表达而发挥作用。To investigate the protective effects and action mechanism of Salvia miltiorrhiza monomerIH764-3 on aristolochic acid（ AA） induced renal tubular epithelial cell injury. Methods The human renal tubular epithelial cell line- HK-C cells were cultured in DMEM medium with 10% fetal calf serum for 72 hours,which were divided into three large groups（ eight small groups） ： control group,the culture solution without AA; AA group,AA was added into culture solution with final concentration of 20 mg / L; observation group,AA and IH764-3（ final concentration of 20 mg / L） and different concentrations of Salvia miltiorrhiza monomer-IH764-3. After cultured for 24 h,48h,72 h,the expression levels of transforming growth factor 1（ TGF- β 1）,tissue inhibitor of matrix metalloproteinase 1（ TIMP-1）,plasminogen activator inhibitor type 1（ PAI-1）,matrix metalloproteinase 2（ MMP-2） in culture medium were detected by enzyme-linked immunosorbent assay（ ELISA） The activity of HK-C cells was measured by MTT assay,and the expression levels of Caspase-3 mRNA were detected by Real-time quantitative PCR. Results After AA stimulation,the expression levels of TGF- β 1,TIMP-1,PAI-1 were increased,however,the expression levels of MMP-2 were obviously decreased（ P〈0. 05）. After intervention by Salvia miltiorrhiza monomer- IH764-3, the expression levels of TGF-β1, TIMP-1 and PAI-1 were decreased, however,the expression levels of MMP-2 were increased,in which the group with IH764-3 80 mg / L had the best effects（ P〈0. 05）. As compared with that in control group,the activity of HK-C cells in AA group and observation group was decreased,however,cell apoptosis rates were increased,moreover,the expression levels of Caspase-3 mRNA were significantly increased（ P〈0. 05）. As compared with that in AA group,the activity of HK-C cells in observation group was obviously increased,however,cell apoptosis rates were decreased,furthermore,the expression levels of Caspase-3 mRNA were significantly decreased（ P�

关 键 词：丹参单体IH764-3 马兜铃酸 人肾小管上皮细胞 细胞凋亡 

分 类 号：R692.6[医药卫生—泌尿科学]