3-甲基腺嘌呤抑制VEGF信号减轻早期糖尿病肾病细胞外基质沉积  

3-Methyladenine alleviates extracellular matrix deposition in early diabetic nephropathy by inhibiting VEGF signaling

作　　者：刘本菊 任海文 王朵 陈建华 刘清云 潘明明 龚权 Liu Benju;Ren Haiwen;Wang Duo;Chen Jianhua;Liu Qingyun;Pan Mingming;Gong Quan(Medical School of Yangtze University,Jingzhou 434023,China)

机构地区：[1]长江大学医学部,荆州434023

出　　处：《中华内分泌代谢杂志》2023年第10期876-881,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金(82270893);长江大学医学科研联合发展基金(WJ2019-16)。

摘　　要：目的探讨3-甲基腺嘌呤(3-MA)对早期糖尿病肾病(DN)细胞外基质沉积的影响及机制。方法采用链脲佐菌素(STZ)诱导1型糖尿病小鼠模型,将小鼠分为溶剂对照组、糖尿病组(STZ组)、3-MA干预组和氯喹(CQ)干预组,每组8只小鼠。干预6周后取双肾,记录肾/体重比,Western印迹检测肾皮质纤连蛋白(fibronectin)、α-平滑肌肌动蛋白(α-SMA)、LC3、Beclin 1、p62和血管内皮生长因子(VEGF)蛋白表达,免疫组化观察肾脏纤连蛋白染色,并对DN基因靶点与3-MA预测基因靶点的共同基因进行生信分析。结果3-MA和CQ对糖尿病小鼠均具有一定降糖作用。与STZ组相比,3-MA组小鼠肾/体重比下降(P<0.05);Western印迹显示,3-MA可减轻糖尿病小鼠肾皮质基质纤连蛋白和α-SMA的表达(P<0.05或P<0.01);免疫组化亦显示,3-MA可减少糖尿病小鼠肾脏纤连蛋白染色。3-MA和CQ均可抑制糖尿病小鼠肾皮质Beclin 1蛋白的表达(均P<0.05),3-MA可增加糖尿病小鼠肾皮质p62的表达(P<0.05)。生信分析结果显示,DN基因靶点与3-MA预测基因靶点的共同基因与VEGF信号有关;Western印迹结果亦显示3-MA可减少糖尿病小鼠肾皮质VEGF表达(P<0.01)。结论3-MA可通过抑制VEGF信号减轻早期DN小鼠肾脏细胞外基质沉积。Objective To investigate the effects of 3-methyladenine(3-MA)on extracellular matrix deposition in early diabetic nephropathy(DN)and its mechanism.Methods A streptozotocin(STZ)-induced type 1 diabetes mouse model was used,and the mice were divided into vehicle control group,diabetes group(STZ group),3-MA group,and chloroquine(CQ)group,8 mice in each group.After 6 weeks of intervention,both kidneys were harvested,and the kidney-to-body weight ratio was recorded.Western blotting was performed to detect protein expressions of renal cortex fibronectin,α-smooth muscle actin(α-SMA),LC3,Beclin 1,p62,and vascular endothelial growth factor(VEGF).Immunohistochemistry was used to observe kidney fibronectin staining.Bioinformatics analysis was conducted on shared genes between diabetic nephropathy(DN)gene targets and 3-MA predicted gene targets.Results Both 3-MA and CQ exhibited certain hypoglycemic effects in diabetic mice.Compared to the STZ group,the kidney-to-body weight ratio decreased in the 3-MA group(P<0.05).Western blotting showed that 3-MA reduced the expression of renal cortex matrix-related proteins fibronectin andα-SMA in diabetic mice(P<0.05 or P<0.01).Immunohistochemistry also revealed that 3-MA reduced fibronectin staining in the kidneys of diabetic mice.Both 3-MA and CQ inhibited the protein expression of renal cortex Beclin 1 in diabetic mice(both P<0.05),while 3-MA increased the expression of renal cortex p62(P<0.05).Bioinformatics analysis indicated a connection between shared genes of DN gene targets and 3-MA predicted gene targets with the VEGF signaling pathway.Western blotting results further showed that 3-MA reduced renal cortex VEGF expression in diabetic mice(P<0.01).Conclusion 3-MA can alleviate extracellular matrix deposition in the kidneys of early DN mice by inhibiting the VEGF signaling pathway.

关 键 词：3-甲基腺嘌呤 糖尿病肾病 细胞外基质 

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