活性维生素D通过维生素D受体调控细胞因子信号转导抑制分子、酪氨酸激酶2/信号转导和转录激活因子3通路在糖尿病肾脏疾病中作用的研究  

Study of the active vitamin D’s role in diabetic kidney diseases through VDR regulation of the SOCS and JAK2/STAT3 pathways

作　　者：周雪 杨孟雪 杨波 孙博文 李飞 龚其海 刘军 查兵兵 ZHOU Xue;YANG Mengxue;YANG Bo(The First Clinical Institute,Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学第一临床学院,563000 [2]复旦大学附属上海市第五人民医院内分泌科 [3]遵义医科大学基础药理教育部重点实验室特色民族药教育部国际合作联合研究室

出　　处：《中国糖尿病杂志》2020年第11期852-860,共9页Chinese Journal of Diabetes

基　　金：国家自然科学基金(81560147);贵州省科技厅支撑计划[黔科合支撑[2017]2884];贵州省科技攻关项目[黔科合SY字(2012)3116号];贵州省科学技术基金项目[黔科合J字LKZ(2013)53号]。

摘　　要：目的探讨活性维生素D(VitD)通过维生素D受体(VDR)调控细胞因子信号转导抑制分子(SOCS)、酪氨酸激酶/信号转导和转录激活因子(JAK/STAT)通路在DKD中的作用及可能机制。方法构建DKD小鼠及沉默VDR基因的DKD小鼠模型,随机分为DKD组、低浓度VitD3干预组(L⁃VitD3)、高浓度VitD3干预组(H⁃VitD3)、慢病毒转染沉默VDR基因表达组(Lenti⁃shVDR)、低浓度VitD3+慢病毒转染组(Lenti⁃shVDR+L⁃VitD3)、高浓度VitD3+慢病毒转染组(Lenti⁃shVDR+H⁃VitD3)。饲养期间每天观察小鼠一般行为学变化,每3~4 d监测BG及体重,第1、4、8周测24小时尿蛋白(24 hUP),HE染色观察肾脏病理改变,Western blot法检测肾脏组织中JAK2、磷酸化JAK2(p⁃JAK2)、STAT3、p⁃STAT3、SOCS1和SOCS3蛋白表达水平。结果与DKD组比较,L⁃VitD3、H⁃VitD3组体重、BG和24 hUP降低(P<0.05),H⁃VitD3组BG和24 hUP低于L⁃VitD3组(P<0.05),Lenti⁃shVDR组24 hUP升高(P<0.05)。DKD组肾小球增大伴系膜基质增生,L⁃VitD3、H⁃VitD3组病变减轻;与DKD组比较,Lenti⁃shVDR组肾小球病变加重,Lenti⁃shVDR+L⁃VitD3、Lenti⁃shVDR+H⁃VitD3组病变较Lenti⁃shVDR组稍减轻。与DKD组比较,L⁃VitD3、H⁃VitD3组p⁃STAT3、SOCS1和SOCS3蛋白表达均降低(P<0.05);JAK2蛋白表达在L⁃VitD3组降低(P<0.05)。与DKD组比较,Lenti⁃shVDR组JAK2、p⁃STAT3、SOCS1和SOCS3蛋白表达均增加(均<0.01)。与Lenti⁃shVDR组比较,Lenti⁃shVDR+H⁃VitD3组JAK2、p⁃STAT3、SOCS1和SOCS3蛋白表达降低(P<0.01)。结论VitD可降低DKD小鼠尿蛋白,改善肾脏病理损伤,延缓肾损害。JAK2/STAT3/SOCS途径可能参与DKD进展,沉默VDR基因后可减弱活性VD对JAK/STAT通路激活的影响,提示活性VitD调控SOCS和JAK/STAT通路可能通过特异性受体VDR介导。Objective To investigate the role and possible mechanism of active vitamin D(VitD)in regulating suppressor of cytokine signaling(SOCS)and Janus kinase signal transducer and activator of tran⁃scription(JAK/STAT)pathways through vitamin D receptor(VDR)in Diabetic kidney disease(DKD).Methods DKD mice model and the DKD mice with silencing VDR gene were constructed and randomly divided into 6 groups:DKD group;low concentration VitD3 intervention group(L⁃VitD3);high concentra⁃tion VitD3 intervention group(H⁃VitD3);lentivirus transfection silencing VDR gene expression group(Lenti⁃shVDR);low concentration VitD3+lentivirus transfection group(Lenti⁃shVDR+L⁃VitD3);and high concentration VitD3+lentivirus transfection group(Lenti⁃shVDR+H⁃VitD3).General behavioral changes were observed every day during feeding.Blood glucose and body weight were monitored every 3~4 days.24⁃hour urine protein(24 hUP)was measured at weeks 1,4 and 8.Hematoxylin⁃Erythrocyte(HE)stained light microscopy was used to observe pathological changes in kidney.Western blot was performed to measure the expression levels of JAK2,phosphorylated JAK2(p⁃JAK2),STAT3,p⁃STAT3,SOCS1,and SOCS3 proteins in kidney tissue.Results Compared with DKD group,body weight,BG and 24 hUP decreased in L⁃VitD3 and H⁃VitD3 groups(P<0.05).BG and 24 hUP were lower in H⁃VitD3 group than in L⁃VitD3 group(P<0.05),and 24 hUP increased in Lenti⁃shVDR group(P<0.05).Glomerular enlargement with mesangial matrix hyperplasia was detected in DKD group,and pathological changes were alleviated in L⁃VitD3 and H⁃VitD3 groups.Compared with DKD group,the glomerular lesions were aggravated in Lenti⁃shVDR group,and the lesions in were slightly relieved Lenti⁃shVDR+L⁃VitD3 and Lenti⁃shVDR+H⁃VitD3 groups as compared with Lenti⁃shVDR group.Compared with DKD group,the protein expressions of p⁃STAT3,SOCS1 and SOCS3 were decreased in L⁃VitD3 and H⁃VitD3 groups(P<0.05).The expression of JAK2 protein decreased in L⁃VitD3 group(P<0.05).Compared with DK

关 键 词：1 25⁃二羟维生素D3 维生素D受体 细胞因子信号转导抑制分子 酪氨酸激酶/信号转导和转录激活因子 糖尿病肾脏疾病 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]