SMYD2特异性抑制剂对高糖环境下大鼠肾成纤维细胞活化的影响及其机制  

Effect of SMYD2 specific inhibitor on high glucose-induced activation of rat renal fibroblasts and its mechanism

作　　者：陈思羽 左思洋 彭睿 李霞 杨元 龙合花 陈敏 杨丹 邹雪 郭兵[3] 刘丽荣 CHEN Siyu;ZUO Siyang;PENG Rui;LI Xia;YANG Yuan;LONG Hehua;CHEN Min;YANG Dan;ZOU Xue;GUO Bing;LIU Lirong(Center for Clinical Laboratories,The Affiliated Hospital of Guizhou Medical University,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,China;Guizhou Precision Medicine Institute,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;Department of Pathophysiology,Guizhou Medical University,Labo-ratory of Pathogenesis Research,Drug Prevention and Treatment of Major Diseases,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州医科大学检验学院,贵州贵阳550004 [2]贵州医科大学附属医院精准医学研究院,贵州贵阳550004 [3]贵州医科大学病理生理学教研室,贵州医科大学重大疾病发病机制研究与药物防治实验室,贵州贵阳550025

出　　处：《中国病理生理杂志》2023年第3期417-424,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81860134,No.32060202);贵州医科大学附属医院博士科研启动基金项目(No.gyfybsky-2021-64);贵州医科大学附属医院2021年度临床研究项目(No.2021-GMHCT-012)。

摘　　要：目的:明确赖氨酸甲基转移酶SMYD2(SET and MYND domain containing 2)特异性抑制剂对体外高糖(HG)环境下大鼠肾成纤维细胞(NRK-49F)活化的影响并探索其可能机制。方法:采用CCK-8法检测SMYD2特异性抑制剂LLY507对体外培养的NRK-49F细胞活力的影响。实验分组为正常糖(NG;含5.5 mmol/L葡萄糖)组、HG(含30 mmol/L葡萄糖)组、NG+LLY507(1.5μmol/L)组和HG+LLY507(1.5μmol/L)组,37℃培养48 h后收集各组细胞蛋白。采用Western blot实验检测SMYD2、组蛋白H3第4位赖氨酸三甲基化(H3K4me3)、纤维连接蛋白(fibronectin)、I型胶原蛋白(Col I)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)、p-Smad3、Smad3、肿瘤坏死因子α(TNF-α)、核因子κB(NF-κB)p65、NF-κB p-p65、白细胞介素6(IL-6)、信号转导及转录激活因子3(STAT3)及p-STAT3蛋白水平。结果:LLY507浓度低于1.6μmol/L对NRK-49F细胞的活力无显著影响;与NG组相比,HG组SMYD2、H3K4me3、fibronectin、Col I、α-SMA、TGF-β1、p-Smad3,TNF-α、NF-κB p-p65、NF-κB p65、IL-6、p-STAT3和STAT3蛋白水平均显著升高(P<0.05);与HG组相比,HG+LLY507组SMYD2、H3K4me3、fibronectin、Col I、α-SMA、TGF-β1、p-Smad3、TNF-α、NF-κB p-p65、NF-κB p65、IL-6、p-STAT3和STAT3蛋白水平均显著降低(P<0.05)。结论:SMYD2特异性抑制剂LLY507可明显抑制HG诱导的NRK-49F细胞活化和细胞外基质分泌,其机制可能与阻断TGF-β1/Smad3、NF-κB和STAT3细胞信号通路激活相关。AIM To determine the specific inhibitory effect of lysine methyltransferase SMYD2(SET and MYND domain containing 2)on high glucose(HG)-induced renal fibroblasts(NRK-49F)and explore its possible mechanism.METHODS The effect of LLY507,an SMYD2 specific inhibitor,on the viability of NRK-49F cells was detected by CCK-8 assay.The cells were divided into normal glucose(NG,5.5 mmol/L glucose)group,HG(30 mmol/L glucose)group,NG+LLY507(1.5μmol/L)group and HG+LLY507(1.5μmol/L)group.The cell lysates of each group were collected after incubation at 37℃for 48 h.Western blot was used to detect the protein expression levels of SMYD2,histone H3 lysine 4 trimethylation(H3K4me3),fibronectin,collagen type I(Col I),α-smooth muscle actin(α-SMA),transforming growth factor-β1(TGF-β1),p-Smad3,Smad3,tumor necrosis factor-α(TNF-α),nuclear factor-κB(NF-κB)p65,NF-κB p-p65,interleukin-6(IL-6),signal transducer and activator of transcription 3(STAT3)and p-STAT3.RESULTS Lower than 1.6μmol/L of LLY507 had no significant effect on the viability of NRK-49F cells.Compared with NG group,the protein expression levels of SMYD2,H3K4me3,fibronectin,Col I,α-SMA,TGF-β1,p-Smad3,TNF-α,NF-κB p-p65,NF-κB p65,IL-6,p-STAT3 and STAT3 were up-regulated in HG group(P<0.05).Compared with HG group,the protein expression levels of SMYD2,H3K4me3,fibronectin,Col I,α-SMA,TGF-β1,p-Smad3,TNF-α,NF-κB p-p65,NF-κB p65,IL-6,p-STAT3 and STAT3 were significantly down-regulated in HG+LLY507 group(P<0.05).CONCLUSION LLY507 inhibits HG-induced secretion of extracellular matrix in NRK-49F cells,which may be attributed to inhibition of TGF-β1/Smad3,NF-κB and STAT3 signaling pathways.

关 键 词：糖尿病肾病 大鼠肾成纤维细胞 SMYD2蛋白 LLY507 细胞外基质 

分 类 号：R587.2[医药卫生—内分泌] R363.2[医药卫生—内科学]