丹参酮II-A对万古霉素所致肾小管上皮细胞KIM-1及TGF-β_1表达的影响  

Effects of Tanshinon on the Expressions of KIM-1, TGF-β_1 in Vancomycin-induced HK-2 Cells

作　　者：陈晓宇[1] 席加喜[1] 刘晓霞[1] 蒙明瑜 叶冬梅[1] CHEN Xiao-yu;XI Jia-xi;LIU Xiao-xia;MENG Ming-yu;YE Dong-mei(Department of medicine, The guangxizhuang autonomous region people＇s hospital, Nanning, Guangxi, 530021, China)

机构地区：[1]广西壮族自治区人民医院药学部,广西南宁530021

出　　处：《现代生物医学进展》2018年第9期1637-1641,共5页Progress in Modern Biomedicine

基　　金：广西医药卫生自筹经费计划课题(Z2016579)

摘　　要：目的:研究丹参酮II-A(TSII-A)对万古霉素(vancomycin,VAN)诱导的人肾近曲小管上皮细胞(HK-2)的存活、氧化应激水平和肾损伤分子1(KIM-1)及转化生长因子-β(TGF-β1)表达的影响。方法:将体外培养的HK-2细胞株接种于6孔培养板,分为空白组、模型组、阳性药物组和TSII-A高、中、低剂量组。加入终浓度为100μg·L-1万古霉素建立VAN损伤HK-2细胞模型,空白组、模型组加入等体积的生理盐水,阳性药物组加入终浓度为2.5 mg·L^(-1)的氨磷汀溶液,TSII-A各剂量组分别予不同浓度的TSII-A处理48 h。进一步通过MTT法测定细胞存活率;裂解细胞取上清液,紫外分光光度法监测细胞内谷胱甘肽(GSH-PX)含量,黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性、硫代巴比妥酸(TBA)法测定丙二醛(MDA)含量、硝酸还原酶法测定一氧化氮(NO)活性;ELISA法测定细胞上清液中KIM-1、TGF-β1的浓度;RT-PCR监测KIM-1、TGF-β1m RNA表达。结果:与空白组比较,模型组的细胞存活率、细胞悬液中的SOD酶活性和GSH-PX含量明显减低,细胞悬液中MDA含量及NO水平明显升高,上清液中KIM-1的浓度明显升高,KIM-1m RNA的相对表达量明显上调(P<0.05);TGF-β1的浓度及其m RNA的表达差异无统计学意义(P>0.05);与模型组比较,阳性组和TSII-A高、中、低剂量组的细胞存活率、细胞悬液中的SOD酶活性和GSH-PX含量明显升高,细胞悬液中MDA含量及NO水平明显降低,上清液中KIM-1的浓度明显降低,KIM-1m RNA的相对表达量明显下调(P<0.05),且作用均有浓度依赖性。TGFβ1的浓度及其m RNA的表达差异无统计学意义(P>0.05)。结论:本研究结果显示TSII-A以剂量依赖性方式减轻VAN所致HK-2细胞损伤,可能机与其减轻氧化应激有关。Objective： To investigate the effects of tanshinon on the expression of KIM-1, TGF-β1 in vancomycin-induced HK-2 cells. Methods： HK-2 cells were randomly divided into six groups： normal saline（NS） group, model group, positive control group, high dose group, middle dose group and low dose group of TSII-A, The model were duplicated with addition of VAN（100 μg·L^（-1））. The model group, positive control group, high dose group, middle dose group and low dose group of TSII-A were treated by saline solutions, Amifostine, panax notoginsenosides（100, 50, 25 mg·L-1）. The cell viability was detected with MTT method, the content of MDA, NO and the activity of SOD, GSH-PX were measured and cell structure was observed. The concentration of KIM-1, TGF-β1 were measured by ELISA, the expressions of KIM-1, TGF-β1 m RNA were measured by RT-PCR. Results： Compared with the model group, the cell viability and activity of SOD, GSH-PX of TSII-A groups and positive control group were significantly increased（P〈0.05）; the content of MDA,NO, KIM-1 were significantly decreased（P〈0.05）; the expression of KIM-1 m RNA were significantly decreased（P〈0.05）; the cell structure was significantly improved, there was no statistical difference（P〈0.05） in the concentration of TGF-β1 and the expression of TGF-β1 m RNA. Conclusions： TSII-A can promote the proliferation of HK-2 cells induced by VAN through relieving the oxidation stress.

关 键 词：丹参酮II-A 万古霉素VAN HK-2细胞 肾损伤分子1(KIM-1) 转化生长因子-β(TGF-β) 

分 类 号：R-33[医药卫生] R285.5