Indoxyl Sulfate Aggravates Podocyte Damage through the TGF-β1/Smad/ROS Signalling Pathway.

IF 2.3 4区医学 Q2 PERIPHERAL VASCULAR DISEASE Kidney & blood pressure research Pub Date : 2024-01-01 Epub Date: 2024-05-10 DOI:10.1159/000538858

Miao Jia, Lihua Lin, Kang Xun, Damei Li, Weijiang Wu, Shaobo Sun, Hong Qiu, Donghua Jin

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Abstract

Introduction: Hyperglycaemia induces the production of a large quantity of reactive oxygen species (ROS) and activates the transforming growth factor β1 (TGF-β1)/Smad signalling pathway, which is the main initiating factor in the formation of diabetic nephropathy. Indoxyl sulphate (IS) is a protein-binding gut-derived uraemic toxin that localizes to podocytes, induces oxidative stress, and inflames podocytes. The involvement of podocyte damage in diabetic nephropathy through the TGF-β1 signalling pathway is still unclear.

Methods: In this study, we cultured differentiated rat podocytes in vitro and measured the expression levels of nephrin, synaptopodin, CD2AP, SRGAP2a, and α-SMA by quantitative real-time PCR (qRT-PCR) and Western blotting after siRNA-mediated TGF-β1 silencing, TGF-β1 overexpression, and the presence of the ROS inhibitor acetylcysteine. We detected the expression levels of nephrin, synaptopodin, CD2AP, SRGAP2a, small mother against decapentaplegic (Smad)2/3, phosphorylated-Smad2/3 (p-Smad2/3), Smad7, NADPH oxidase 4 (NOX4), and ROS levels under high glucose (HG) and IS conditions.

Results: The results indicated that nephrin, synaptopodin, CD2AP, and SRGAP2a expressions were significantly upregulated, and α-SMA expression was significantly downregulated in the presence of HG under siRNA-mediated TGF-β1 silencing or after the addition of acetylcysteine. However, in the presence of HG, the expressions of nephrin, synaptopodin, CD2AP, and SRGAP2a were significantly downregulated, and the expression of α-SMA was significantly upregulated with the overexpression of TGF-β1. IS supplementation under HG conditions further significantly reduced the expressions of nephrin, synaptopodin, CD2AP, and SRGAP2a; altered the expressions of Smad2/3, p-Smad2/3, Smad7, and NOX4; and increased ROS production in podocytes.

Conclusion: This study suggests that IS may modulate the expression of nephrin, synaptopodin, CD2AP, and SRGAP2a by regulating the ROS and TGF-β1/Smad signalling pathways, providing new theoretical support for the treatment of diabetic nephropathy.

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硫酸吲哚酯通过 TGF-β1/Smad/ROS 信号通路加重荚膜损伤。

引言高血糖诱导产生大量活性氧（ROS），并激活转化生长因子β1（TGF-β1）/Smad 信号通路，这是糖尿病肾病形成的主要启动因素。硫酸吲哚酯（IS）是一种与蛋白质结合的肠源性尿毒症毒素，可定位到荚膜细胞，诱导氧化应激并使荚膜细胞发炎。目前尚不清楚荚膜细胞损伤是通过 TGF-β1/) 信号通路参与糖尿病肾病的：本研究中，我们在体外培养了分化的大鼠荚膜细胞，并在 siRNA 介导的 TGF-β1 沉默、TGF-β1 过表达和 ROS 抑制剂乙酰半胱氨酸存在的情况下，通过实时定量 PCR (qRT-PCR) 和 Western 印迹检测了 nephrin、突触素、CD2AP、SRGAP2a 和 α-SMA 的表达水平。我们检测了高糖（HG）和IS条件下肾素、突触素、CD2AP、SRGAP2a、Smad2/3中SRGAP2a、磷酸化Smad2/3（p-Smad2/3）、Smad7、NADPH氧化酶4（NOX4）和ROS的表达水平：结果表明，在 siRNA 介导的 TGF-β1 沉默或添加乙酰半胱氨酸的条件下，肾素、突触素、CD2AP 和 SRGAP2a 的表达明显上调，α-SMA 的表达明显下调。然而，在 HG 存在的条件下，肾素、突触素、CD2AP 和 SRGAP2a 的表达明显下调，而 α-SMA 的表达则随着 TGF-β1 的过表达而明显上调。在 HG 条件下补充 IS 会进一步显著降低 nephrin、突触素、CD2AP 和 SRGAP2a 的表达，改变 Smad2/3、p-Smad2/3、Smad7 和 NOX4 的表达，并增加荚膜细胞中 ROS 的产生：本研究表明，IS 可通过调节 ROS 和 TGF-β1/Smad 信号通路来调节肾素、突触素、CD2AP 和 SRGAP2a 的表达，为糖尿病肾病的治疗提供了新的理论支持。

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来源期刊

Kidney & blood pressure research 医学-泌尿学与肾脏学

CiteScore

4.80

自引率

3.60%

发文量

审稿时长

6-12 weeks

期刊介绍： This journal comprises both clinical and basic studies at the interface of nephrology, hypertension and cardiovascular research. The topics to be covered include the structural organization and biochemistry of the normal and diseased kidney, the molecular biology of transporters, the physiology and pathophysiology of glomerular filtration and tubular transport, endothelial and vascular smooth muscle cell function and blood pressure control, as well as water, electrolyte and mineral metabolism. Also discussed are the (patho)physiology and (patho) biochemistry of renal hormones, the molecular biology, genetics and clinical course of renal disease and hypertension, the renal elimination, action and clinical use of drugs, as well as dialysis and transplantation. Featuring peer-reviewed original papers, editorials translating basic science into patient-oriented research and disease, in depth reviews, and regular special topic sections, ''Kidney & Blood Pressure Research'' is an important source of information for researchers in nephrology and cardiovascular medicine.