丹酚酸B对高糖诱导的大鼠肾小管上皮细胞转分化的影响及其机制研究  

Effect of salvianolic acid B on high-glucose induced renal tubular epithelial-mesenchymal transition in rats and its mechanism

作　　者：孙兰 田平平[1,2] 张帆 肖瑛 郭兵 SUN Lan;TIAN Ping-ping;ZHANG Fan;XIAO Ying;GUO Bing(Department of Pathophysiology,School of Basic Medical Sciences,Gaizhou Medlical University,Guiyang 550025,China;Guizhou Prorincial Key Laboratory of Pathogenesis&Drug Research on Common Chronic Diseases,Guizhou Medical Uraisersity,Guiyang 550025,China)

机构地区：[1]贵州医科大学基础医学院病理生理学教研室,贵州贵阳550025 [2]贵州医科大学贵州省常见慢性疾病发病机制及药物研究重点实验室,贵州贵阳550025

出　　处：《中国中药杂志》2020年第16期3922-3930,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(N81860656);贵州省自然科学基金项目贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]191);贵州省中医药管理局中医药、民族医药科学技术研究基金项目(QZYY-2016-077)。

摘　　要：观察丹参的主要成分丹酚酸B(salvianolic acid B,Sal B)对高糖诱导的大鼠肾小管上皮细胞向间充质细胞转分化(epithelial-mesenchymal transition, EMT)的影响,探讨其防治糖尿病肾病(diabetic nephropathy,DN)的可能机制。采用体外培养大鼠近端肾小管上皮NRK-52E细胞,随机分为对照组(NG)、高糖组(HG)、高糖+10μmol·L^-1 Sal B组(Sal B),上述3组分别设6,12,24,48 h 4个时间点进行动态观察;高糖+Sal B不同浓度(1,5,10μmol·L^-1)组、高糖+5.0μmol·L^-1吡格列酮对照组(pioglitazone,PIO)、高糖+10μmol·L^-1 Sal B+5.0μmol·L^-1 GW9662对照组(Sal B+GW9662)。采用Western blot法检测PPARγ,PTEN,α-SMA,E-cadherin的表达变化以及PI3K/Akt通路的变化;Real-time PCR法检测PPARγ,PTEN mRNA的表达水平。结果显示,与NG组相比,HG组PPARγ,PTEN mRNA和蛋白表达水平均逐渐降低,α-SMA,p-Akt(Thr308)蛋白表达水平逐渐增高,E-cadherin蛋白表达水平逐渐降低,呈时间依赖性;与HG组相比,高糖+Sal B不同浓度组随Sal B剂量增加,PPARγ,PTEN mRNA和蛋白表达水平逐渐增高,α-SMA,p-Akt(Thr308)蛋白表达水平逐渐降低,E-cadherin蛋白表达水平逐渐增高,呈剂量依赖性(P<0.05),但1μmol·L^-1浓度的Sal B对PPARγ mRNA和蛋白、PTEN mRNA表达的影响没有显著性差异;与HG组相比,Sal B动态观察组PPARγ mRNA(除6 h)和蛋白(除6 h),PTEN mRNA(除6 h)和蛋白(除6,12 h)表达水平明显增高,α-SMA,p-Akt(Thr308)(除6 h)蛋白表达水平明显降低,E-cadherin蛋白表达水平明显增高(P<0.05);与HG组相比,Sal B和PIO显著增强PPARγ,PTEN mRNA和蛋白表达水平,增高E-cadherin蛋白表达水平,降低α-SMA,p-Akt(Thr308)蛋白表达水平(P<0.05),各用药组之间差异无统计学意义;而Sal B+GW9662对照组PPARγ和PTEN的mRNA和蛋白,E-cadherin、α-SMA和p-Akt(Thr308)蛋白表达水平与高糖组比较没有统计学意义,Sal B的作用效应被PPARγ拮抗剂GW9662阻断。结果表明,丹酚酸B可抑制高糖诱导的NRK-52E细胞发�The aim of this paper was to observe the effect of salvianolic acid B(Sal B) on high-glucose induced renal tubular epithelial-mesenchymal transition(EMT) in rats, and to explore its possible mechanisms of prevention and treatment of diabetic nephropathy. The rat renal tubular epithelial NRK-52 E cells were cultured in vitro. The cells were divided into control group, high glucose group, high glucose+10 μmol·L^-1Sal B group(Sal B), the above 3 groups were set at 6, 12, 24 and 48 h for dynamic observation;high glucose+Sal B different concentration(1, 5, 10 μmol·L^-1) groups, high glucose+5.0 μmol·L^-1 pioglitazone group, high glucose+10 μmol·L^-1Sal B+5 μmol·L^-1GW9662 group. The protein expression levels of PPARγ, PTEN, α-SMA, E-cadherin and PI3 K/Akt signaling molecules were determined by Western blot. The mRNA expression of PPARγ and PTEN were detected by Real-time PCR. The viabi-lity of NRK52 E cells was determined by MTT assay. The results showed that as compared with control group, the mRNA and protein expression levels of PPARγ and PTEN in high glucose group gradually reduced, the protein expression levels of α-SMA and p-Akt(Thr308)gradually increased, and the protein expression of E-cadherin gradually reduced(P<0.05). As compared with high glucose group, when increases in Sal B doses, the mRNA and protein expression levels of PPARγ, PTEN in high glucose + different concentrations of Sal B groups gradually increased, the protein expression levels of α-SMA and p-Akt(Thr308) gradually reduced, and the protein expression of E-cadherin gradually increased(P<0.05), however, the effect of 1 μmol·L^-1concentration of Sal B on the expression of PPARγ mRNA and protein and PTEN mRNA was not significantly different. As compared with high glucose group, the mRNA and protein expression levels of PPARγ mRNA(except 6 h) and protein(except 6 h), PTEN mRNA(except 6 h) and protein(except 6, 12 h) kept increasing, the protein expression levels of α-SMA and p-Akt(Thr308)(except 6 h) continued to reduce, th

关 键 词：丹参 丹酚酸B PPARΓ PTEN 上皮-间充质转分化 

分 类 号：R285.5[医药卫生—中药学]