Vitamin D/vitamin D receptor/Atg16L1 axis maintains podocyte autophagy and survival in diabetic kidney disease.
Vitamin D/vitamin D receptor/Atg16L1 axis maintains podocyte autophagy and survival in diabetic kidney disease.
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维生素D/维生素D受体/Atg16L1轴维持糖尿病肾病足细胞自噬和存活。
DOI：
10.1080/0886022x.2022.2063744
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发表时间：
2022-12
期刊：
影响因子：
3
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中科院分区：
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探讨维生素D/维生素D受体(VDR)/Atg16L1信号通路对糖尿病肾病足细胞自噬及存活的影响。通过腹腔注射链脲佐菌素（STZ） （60 mg/kg）诱导糖尿病大鼠模型，并分别灌胃和不灌胃0.1 μg/kg/d活性维生素D3 （aVitD3; 1,25- OH维生素D3）和肾脏组织，采用组织病理学和免疫组化评价。在无100nmol /L骨化三醇和无100nmol /L骨化三醇的高血糖条件下培养小鼠足细胞MPC-5，观察足细胞损伤和自噬情况。用细胞计数试剂盒-8 （CCK-8）分析细胞存活率，用mRFP-GFP-LC3自噬报告基因转导后测定自噬小体的数量。Western blotting检测自噬相关蛋白（LC3-II、beclin-1、Atg16L1）和足细胞相关蛋白（nephrin、podocin、synaptopodin、desmin）的表达。糖尿病肾病组VDR表达减少，自噬减少。骨化三醇治疗可抑制糖尿病大鼠肾脏损伤，降低高糖诱导的足细胞损伤。在机制上，Atg16L1被鉴定为VDR的功能靶点，sirna介导的VDR和Atg16L1的敲低阻断了aVitD3对足细胞损伤的保护作用。自噬保护足细胞免受DN损伤，并受VitD3/VDR信号和Atg16L1表达的下游调节。
To investigate the effect of vitamin D/vitamin D receptor (VDR)/Atg16L1 signaling on podocyte autophagy and survival in diabetic nephropathy. Diabetic rat models were induced by intraperitoneal injection of streptozotocin (STZ) (60 mg/kg) and treated with and without gavage of 0.1 μg/kg/d active vitamin D3 (aVitD3; 1,25- OH vitamin D3) and kidney tissues assessed by histopathology and immunohistochemistry. The murine podocyte cell line MPC-5 was cultured under hyperglycemic conditions in the absence or presence of 100 nmol/L calcitriol to investigate podocyte injury and autophagy. Cell survival rates were analyzed using Cell Counting Kit-8 (CCK-8) assays and the numbers of autophagosomes were determined after transduction with the mRFP-GFP-LC3 autophagy reporter construct. The expression of autophagy-related proteins (LC3-II, beclin-1, Atg16L1) and podocyte-related proteins (nephrin, podocin, synaptopodin, and desmin) was determined by Western blotting. VDR expression and autophagy were decreased in diabetic nephropathy. Calcitriol treatment repressed renal injury in rat diabetic kidneys and reduced high glucose-induced damage to cultured podocytes. Mechanistically, Atg16L1 was identified as a functional target of VDR, and siRNA-mediated knockdown of VDR and Atg16L1 blocked the protective effects of aVitD3 against podocyte damage. Autophagy protects podocytes from damage in DN and is modulated by VitD3/VDR signaling and downstream regulation of Atg16L1 expression.