TXNIP 通过 AMPK 通路部分介导肾小球系膜细胞的氧化应激反应。
TXNIP mediated the oxidative stress response in glomerular mesangial cells partially through AMPK pathway.
机构信息
Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, China.
Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, China.
出版信息
Biomed Pharmacother. 2018 Nov;107:785-792. doi: 10.1016/j.biopha.2018.08.067. Epub 2018 Aug 22.
BACKGROUND
Thioredoxin-interacting protein (TXNIP) plays an important role in the development of diabetic nephropathy. In the present study, we investigated role of TXNIP on oxidative stress in glomerular mesangial cells (GMCs) cultured in high glucose or normal glucose, and explored the potential mechanism related to TXNIP as well.
METHODS
Oxidative stress in GMCs under high or normal glucose was detected. TXNIP knockdown by specific siRNA or over expression by pcDNA3.0-TXNIP vector was performed to evaluate the role of TXNIP on injury of GMCs caused by oxidative stress. Activator of AMPK AICAR and AMPK inhibitor Compound C were treated the GMCs. Reactive oxygen species (ROS) and mitochondrial membrane potential were detected by flow cytometry. Activities of superoxide dismutase (SOD) and superoxide dismutase (CAT) were measured by ELISA. Activity of thioredoxin (Trx) was determined using Trx activity assay kit. mRNA expression of AMPK, TXNIP, Trx1 and Trx2 were tested by qRT-PCR. Expressions of P-AMPK, TXNIP and fibronectin proteins were detected by Western blotting.
RESULTS
High glucose induced the increase of ROS level, activation of TXNIP, but restricted mitochondrial membrane potential and activities of p-AMPK, SOD and CAT, and Trx. TXNIP siRNA and AICAR inhibited high glucose-induced oxidative stress response in GMCs and fibronectin expression, but promoted cell viability. In contrast, pcDNA3.0-TXNIP and Compound C increased oxidative stress response in normal glucose cultured GMCs, but decreased cell viability. The combined effect of TXNIP siRNA and AICAR on the inhibition of oxidative stress was obviously stronger than that of single use of TXNIP siRNA.
CONCLUSION
TXNIP facilitates the oxidative stress response in GMCs partially through AMPK pathway, which may provide potential therapeutic target for diabetic nephropathy treatment.
背景
硫氧还蛋白相互作用蛋白（TXNIP）在糖尿病肾病的发展中起着重要作用。本研究旨在探讨 TXNIP 在高糖或正常糖培养的肾小球系膜细胞（GMC）中氧化应激中的作用，并探讨与 TXNIP 相关的潜在机制。
方法
检测高糖或正常糖培养的 GMC 中的氧化应激。通过特异性 siRNA 或 pcDNA3.0-TXNIP 载体过表达来评估 TXNIP 在氧化应激引起的 GMC 损伤中的作用。用 AMPK 激活剂 AICAR 和 AMPK 抑制剂 Compound C 处理 GMC。用流式细胞术检测活性氧（ROS）和线粒体膜电位。用 ELISA 法测定超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。用硫氧还蛋白（Trx）活性测定试剂盒测定 Trx 活性。用 qRT-PCR 检测 AMPK、TXNIP、Trx1 和 Trx2 的 mRNA 表达。用 Western blot 检测 P-AMPK、TXNIP 和纤维连接蛋白蛋白的表达。
结果
高糖诱导 ROS 水平升高，TXNIP 激活，但限制了线粒体膜电位和 p-AMPK、SOD 和 CAT 及 Trx 的活性。TXNIP siRNA 和 AICAR 抑制了高糖诱导的 GMC 氧化应激反应和纤维连接蛋白表达，但促进了细胞活力。相反，pcDNA3.0-TXNIP 和 Compound C 增加了正常糖培养的 GMC 中的氧化应激反应，但降低了细胞活力。TXNIP siRNA 和 AICAR 的联合作用对氧化应激的抑制作用明显强于单独使用 TXNIP siRNA。
结论
TXNIP 通过 AMPK 途径促进 GMC 中的氧化应激反应，这可能为糖尿病肾病的治疗提供潜在的治疗靶点。
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