益糖康对高糖诱导人肾小管上皮HK-2细胞上皮间质转化的影响  

Effects of Yitangkang on epithelial mesenchymal transformation of human renal tubular epithelial cells HK-2 induced by high glucose

作　　者：郭隽馥[1] 杨宇峰[1] 石岩[1] GUO Jun-fu;YANG Yu-feng;SHI Yan(Teaching Experiment Center,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)

机构地区：[1]辽宁中医药大学教学实验中心,沈阳110847

出　　处：《中华中医药杂志》2021年第3期1735-1739,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.81803855);中国博士后科学基金面上项目(No.2019M651141);辽宁省博士科研启动基金计划项目(No.2019-BS-166);沈阳市中青年科技创新人才支持计划项目(No.RC190078);辽宁省“兴辽英才计划”青年拔尖人才资助项目(No.XLYC1807145)。

摘　　要：目的:探讨益糖康是否通过抑制肾小管上皮细胞发生上皮-间质转化(EMT)进而减轻糖尿病对肾脏的早期损伤。方法:40只SD雄性大鼠随机分成生理盐水组(1mL/100g)、益糖康组(2.10g/100g)、二甲双胍组(9.11g/100g)、二甲双胍+益糖康组(4.55g/100g二甲双胍+1.05g/100g益糖康煎剂),每组10只,分别给予相应药物灌胃,连续5d,腹主动脉取血,分离上清液。体外实验分组:NS组(10%生理盐水组血清)、YTK组(10%益糖康组血清)、Metformin组(10%二甲双胍组血清)和M+YTK组(10%二甲双胍+益糖康组血清)。高糖刺激人肾小管上皮HK-2细胞24h后,分别采用划痕实验、Transwell实验、Real-time PCR和Western Blot法检测细胞迁移力、体外侵袭力以及EMT相关基因表达水平的改变。结果:YTK组细胞划痕区的未汇合率显著高于NS组(P<0.05);YTK组穿膜细胞数显著低于NS组(P<0.05);M+YTK组穿膜细胞数显著低于Metformin组(P<0.05);与NS组比较,YTK、Metformin及M+YTK组细胞中E-cadherin mRNA表达水平显著升高(P<0.05,P<0.01);而Vimentin和α-SMA mRNA表达水平均显著降低(P<0.01),其中YTK组和Metformin组水平相当,而M+YTK组中Vimentin和α-SMA mRNA表达水平显著低于Metformin组(P<0.05)。结论:益糖康可能是通过抑制肾小管细胞发生EMT进而减轻糖尿病对肾脏的早期损伤,且中药复方益糖康联合西药二甲双胍对肾小管上皮细胞的EMT抑制效果较单独使用二甲双胍更为显著。Objective: To investigate whether Yitangkang can reduce the early renal injury caused by diabetes by inhibiting the epithelial-mesenchymal transition(EMT) of renal tubular epithelial cells. Methods: A total of 40 male SD rats were randomly divided into normal saline group(1 mL/100 g), Yitangkang group(2.10 g/100 g), metformin group(9.11 g/100 g) and metformin+Yitangkang group(4.55 g/100 g metformin+1.05 g/100 g Yitangkang Decoction), with 10 rats in each group by gavage for 5 days. Blood was collected from abdominal aorta and supernatant was separated by centrifugation. The in vitro experiment groups: NS group(10%normal saline group serum), YTK group(10%Yitangkang group serum), Metformin group(10% metformin group serum) and M+YTK group(10%metformin+Yitangkang group serum). Human renal tubular epithelial HK-2 cells were stimulated by high glucose for 24 h, and the changes of cell migration capacity, invasion ability in vitro and expression level of EMT-related genes were detected by wounding healing assay, Transwell assay, Real-time PCR and Western Blot, respectively. Results: The non-confluence rate of wounding healing area in YTK group was significantly higher than that in NS group(P<0.05). Compared with NS group, the number of transmembrane cells in YTK group was significantly reduced(P<0.05). The number of transmembrane cells in M+YTK group was significantly lower than that in Metformin group(P<0.05). Compared with NS group, the mRNA expression levels of E-cadherin in YTK, Metformin and M+YTK groups were significantly increased(P<0.05, P<0.01). The mRNA expression levels of Vimentin and α-SMA in the YTK, Metformin and M+YTK groups were significantly lower than those in the NS group(P<0.01), and the expression levels of Vimentin and α-SMA in the YTK group were similar to those in the Metformin group. The mRNA expression levels of Vimentin and α-SMA in the M+YTK group were significantly lower than those in the Metformin group(P<0.05). Conclusion: Yitangkang may reduce the early renal injury caused by diabetes

关 键 词：益糖康 糖尿病 肾小管上皮细胞 HK-2 上皮-间质转化 二甲双胍 

分 类 号：R285.5[医药卫生—中药学]