组蛋白甲基转移酶SMYD2对巨噬细胞-肌成纤维细胞转化促进糖尿病肾病肾纤维化的影响  

Effect of histone methyltransferase SMYD2 on macrophage-myofibro-blast transition-promoted renal fibrosis in diabetic kidney disease

作　　者：杨元 彭睿 刘泽莹 邹雪 李霞[3] 元辉雄 龙合花 王腾 岑明杰 郭兵[3] 朱丽英 刘丽荣[1,2,3] YANG Yuan;PENG Rui;LIU Zeying;ZOU Xue;LI Xia;YUAN Huixiong;LONG Hehua;WANG Teng;CEN Mingjie;GUO Bing;ZHU Liying;LIU Lirong(Center for Clinical Laboratories,The Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,China;Key Laboratory of Kidney Disease Pathogenesis Research and Transformation Application,Guizhou Medical University,Guiyang 561113,China;Center for Clinical Medical Research,The Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [2]贵州医科大学医学检验学院,贵州贵阳550004 [3]贵州医科大学肾脏病发病机制及转化应用重点实验室,贵州贵阳561113 [4]贵州医科大学附属医院临床医学研究中心,贵州贵阳550004

出　　处：《中国病理生理杂志》2025年第2期239-249,共11页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81860134);贵州省科技计划项目(No.黔科合基础-ZK[2023]一般377)。

摘　　要：目的:探讨组蛋白甲基转移酶SMYD2(SET and MYND domain containing 2)在巨噬细胞转化促进糖尿病肾病(diabetic kidney disease,DKD)肾纤维化中的作用。方法:(1)C57BL/6J小鼠给予55 mg/kg链脲佐菌素腹腔注射,建立糖尿病模型。实验分为正常对照(normal control,NC)组(n=5)、糖尿病20周组(n=5)、糖尿病28周组(n=5)和糖尿病36周组(n=5)。生化分析仪检测小鼠血糖(blood glucose,BG)、血清肌酐(serum creatinine,SCr)和血尿素氮(blood urea nitrogen,BUN);苏木精-伊红(hematoxylin and eosin,HE)和Masson染色观察小鼠肾组织形态学及纤维化改变情况;Western blot检测SMYD2、组蛋白H3第4位赖氨酸三甲基化(histone H3 lysine 4 trimethylation,H3K4me3)、精氨酸酶1(arginase-1)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)、I型胶原(collagen type I,Col I)和α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)的蛋白水平;免疫荧光染色观察F4/80、α-SMA、SMYD2、CD86、CD206和CD163蛋白的定位及表达情况。(2)体外培养小鼠单核巨噬细胞(RAW264.7),分为正常糖(normal glucose,NG)+敲减对照(negative control siRNA,siNC)组、高糖(high glucose,HG)+siNC组、NG+SMYD2敲减(SMYD2 siRNA,siSMYD2)组和HG+siSMYD2组。Western blot检测相关蛋白表达情况。结果:(1)与NC组相比,糖尿病28周和糖尿病36周组的BG、SCr和BUN均增高(P<0.05),肾组织可见肾小管萎缩、扩张及胶原纤维沉积,H3K4me3、arginase-1、MMP9、Col I和α-SMA蛋白表达上调(P<0.05)。CD86、CD206、CD163和F4/80主要表达在肾小管间质巨噬细胞,α-SMA主要表达在肾间质,SMYD2主要表达在肾小管上皮细胞及肾间质。(2)与NG+siNC组相比,HG+siNC组SMYD2、H3K4me3、arginase-1、CD163、Col I、α-SMA、转化生长因子β1(transforming growth factor-β1,TGF-β1)和p-Smad3蛋白水平显著升高(P<0.05);敲减SMYD2后上述指标水平降低(P<0.05)。结论:SMYD2可通过巨噬细胞-肌成纤维细胞转化促进DKD肾纤维化,其机制可能与调控TGF-β1AIM:This study aims to investigate the role of histone methyltransferase SET and MYND domain containing 2(SMYD2)in facilitating renal fibrosis through the macrophage-myofibroblast transition in diabetic kidney dis-ease(DKD).METHODS:(1)C57BL/6J mice were intraperitoneally administered 55 mg/kg of streptozotocin to induce diabetes mellitus(DM).The experimental groups were categorized as follows:normal control,DM(20 weeks),DM(28 weeks),and DM(36 weeks).Blood glucose(BG),serum creatinine(SCr)and blood urea nitrogen(BUN)levels were determined using a biochemical analyzer.Hematoxylin-eosin(HE)staining and Masson staining were performed to assess morphological and fibrotic changes in renal tissues.Western blot analysis was used to measure the protein levels of SMYD2,histone H3 lysine 4 trimethylation(H3K4me3),arginase-1,matrix metalloproteinase 9(MMP9),collagen type I(Col I)andα-smooth muscle actin(α-SMA).Immunofluorescence staining was conducted to examine the localization and expression of F4/80,α-SMA,SMYD2,CD86,CD206 and CD163.(2)Mouse monocyte/macrophage RAW264.7 cells were cultured in vitro and assigned to groups as follows:normal glucose(NG)+negative control siRNA(siNC),high glucose(HG)+siNC,NG+SMYD2 siRNA(siSMYD2),and HG+siSMYD2.Western blot analysis was used to assess the expression of relevant proteins.RESULTS:(1)Compared with normal control group,the levels of BG,SCr and BUN were significantly elevated in DM(28 weeks)and DM(36 weeks)groups(P<0.05).Renal tissue exhibited tubular atro-phy,dilation,and collagen fiber deposition.The levels of H3K4me3,arginase-1,MMP9,Col I andα-SMA proteins were up-regulated(P<0.05).The CD86,CD206,CD163 and F4/80 were primarily localized in the interstitial macrophages of the renal tubules,α-SMA was predominantly detected in the renal interstitium,and SMYD2 was mainly expressed in renal tubular epithelial cells and the renal interstitium.(2)Compared with NG+siNC group,the protein levels of SMYD2,H3K4me3,arginase-1,CD163,Col I,α-SMA,transforming growth factor-β1(TGF-β1)and p

关 键 词：甲基转移酶 SMYD2蛋白 糖尿病肾病 肾纤维化 巨噬细胞 肌成纤维细胞 

分 类 号：R587.2[医药卫生—内分泌] R363.2[医药卫生—内科学] Q291[医药卫生—临床医学]