NOX1 abet mesangial fibrogenesis via iNOS induction in diabetes
NOX1 abet mesangial fibrogenesis via iNOS induction in diabetes
复制标题
NOX1 通过 iNOS 诱导糖尿病促进系膜纤维化
DOI：
10.1007/s11010-013-1733-4
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发表时间：
2013-10-01
影响因子：
4.3
通讯作者：
Li, Jing
中科院分区：
文献类型：
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作者：
Gao, Ling;Huang, Weilu;Li, Jing
NADPH氧化酶（NOX）和诱导型一氧化氮合酶（iNOS）是肾脏活性氧的主要来源。然而，它们在氧化应激中的相互作用和对糖尿病肾病期间肾纤维化的贡献尚未研究。人肾小球系膜细胞分别用正常葡萄糖（5.6mmol/L）、高糖（30 mmol/L）及AGE（200 mg/L）处理。通过免疫印迹法检测NOX 1、NOX 2、NOX 4和iNOS的蛋白表达。利用特异性RNAi技术沉默NOX基因，研究NOX与iNOS在糖尿病环境中的相互作用。超氧化物（O·−）和过氧亚硝酸盐（ONOO·−）的产生分别由二氢乙锭和羟苯基荧光素进行评估。生化法测定纤维化因子。糖尿病环境（高糖30 mmol/L + AGE 200 mg/L）中超氧化物歧化酶、过氧亚硝基阴离子、TGF-β和纤维连接蛋白的产生以及NOX 1、NOX 2、NOX 4和iNOS的蛋白表达增加。然而，用1400 W或iNOSRNAi消除iNOS诱导将使过氧亚硝酸盐、TGF-β和纤连蛋白的产生完全恢复到基础水平，并减弱超氧化物的产生。此外，NOX 1抑制不仅阻止了iNOS诱导，而且还消除了iNOS诱导引起的变化，如系膜纤维化。
Both NADPH oxidase (NOX) and inducible nitric oxide synthase (iNOS) are the main sources of reactive oxygen species in kidney. However, their interactions in oxidative stress and contributions to kidney fibrosis during diabetic nephropathy have not been studied. Human mesangial cells were treated with normal glucose (5.6 mmol/L), high glucose (30 mmol/L) in the presence or absence of AGE (200 mg/L). Protein expressions of NOX1, NOX2, NOX4, and iNOS were examined by immunoblotting. NOX was genetically silenced with specific RNAi to study the interactions between NOX and iNOS in diabetic milieu. Superoxide (O·−) and peroxynitrite (ONOO·−) productions were assessed by dihydroethidium and hydroxyphenyl fluorescein, respectively. Fibrotic factors were determined by biochemistry assay. Superoxide, peroxynitrite, TGF-β, and fibronectin productions as well as the protein expressions of NOX1, NOX2, NOX4, and iNOS were increased in the diabetic milieu (high glucose 30 mmol/L plus AGE 200 mg/L). However, abolishment of iNOS induction with 1400W or iNOS RNAi would restore peroxynitrite, TGF-β, and fibronectin productions completely to basal level and attenuate superoxide production. Moreover, NOX1 inhibition not only prevented iNOS induction but also abrogated changes consequent to iNOS induction such as mesangial fibrogenesis.