Macrophage-stimulating protein attenuates hydrogen peroxide-induced apoptosis in human renal HK-2 cells
Macrophage-stimulating protein attenuates hydrogen peroxide-induced apoptosis in human renal HK-2 cells
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DOI：
10.1016/j.ejphar.2013.05.006
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发表时间：
2013-09-05
影响因子：
5
通讯作者：
Kim, Soo Wan
中科院分区：
文献类型：
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作者：
Lee, Ko Eun;Kim, Eun Young;Kim, Soo Wan
巨噬细胞刺激蛋白（Macrophage-stimulating protein，MSP）及其受体（recepteur d 'origine nantais，罗恩）在细胞增殖和迁移中起重要作用。我们研究了MSP在过氧化氢（H2 O2）诱导的肾小管细胞凋亡中的作用。用不同浓度的MSP与H2 O2孵育人肾近端小管（HK-2）细胞24 h，MTT法检测细胞活力。半定量免疫印迹法检测Bax、Bcl-2、caspase-3、丝裂原活化蛋白激酶（MAPK）、磷脂酰肌醇-3-激酶（PI 3 K）/Akt和核因子-κ B（NF-κ B）的蛋白表达。用异硫氰酸荧光素标记的膜联蛋白V蛋白和碘化丙啶对HK-2细胞进行染色后，通过流式细胞术分析来评估细胞凋亡。H2 O2处理降低HK-2细胞的细胞活力，这被MSP预处理所抵消。H2 O2处理诱导Bax/Bcl-2比率增加，切割的caspase-3和浓缩的核的数量，这也被MSP抵消。流式细胞仪分析显示H2 O2诱导的细胞凋亡，并通过MSP处理的预防。MSP可抑制磷酸化p38 MAPK蛋白表达的增加，而磷酸化细胞外信号调节激酶和c-Jun-N-末端激酶的表达无明显变化。H2 O2诱导NF-κ B活化和I κ B-α降解，但增加的核NF-κ B活化被MSP或p38 MAPIK抑制剂抵消。H2 O2处理降低磷酸化PI 3 K和磷酸化Akt的表达，MSP预处理可逆转此作用。这些结果表明MSP通过调节p38和NF-κ B B以及PI 3 K/Akt信号通路来减弱H2 O2诱导的HK-2细胞凋亡。（C）.© 2013 Elsevier B. V.版权所有
Macrophage-stimulating protein (MSP) and its receptor, recepteur d'origine nantais (RON), play an important role in cell proliferation and migration. We have investigated the role of MSP in hydrogen peroxide (H2O2)-induced renal tubular apoptosis. Human renal proximal tubular (HK-2) cells were incubated with H2O2 for 24 h in the presence of different concentrations of MSP, and cell viability was measured by MIT assay. The protein expression of Bax, Bcl-2, caspase-3, mitogen-activated protein kinases (MAPKs), phosphatidylinositol-3-kinase (PI3K)/Akt, and nuclear factor-kappa B (NF-kappa B) was determined by semiquantitative immunoblotting. Apoptosis was assessed by flow cytometry analysis after HK-2 cells were stained with fluorescein isothiocyanate-conjugated annexin V protein and propidium iodide. H2O2 treatment decreased cell viability in HK-2 cells; this was counteracted by MSP pretreatment H2O2 treatment induced an increased ratio of Bax/Bcl-2, cleaved caspase-3, and the number of condensed nuclei, which was also counteracted by MSP. Flow cytometry analysis showed H2O2 induced apoptosis, and its prevention by MSP treatment. Increased protein expression of phospho-p38 MAPK was attenuated by MSP, while phospho-extracellular signal regulated kinase and c-Jun-N-terminal kinase were not affected. H2O2 induced NF-kappa B activation and I kappa B-alpha degradation, but the increased nuclear NF-kappa B activation was counteracted by MSP or by a p38 MAPIK inhibitor. H2O2 treatment decreased expression of phospho-PI3K and phospho-Akt, which was reversed by MSP pretreatment. These findings suggest that MSP attenuates H2O2-induced apoptosis in HK-2 cells by modulating the p38 and NF-kappa B, as well as PI3K/Akt, signaling pathways. (C). 2013 Elsevier B.V. All rights reserved