Ripasudil对PDGF⁃BB诱导人肺动脉平滑肌细胞增殖和迁移的影响
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国家科技重大专项(2015ZX10003001);江苏省卫生厅重点项目(H201601)


Effects of ripasudil on proliferation and migration of human pulmonary arterial smooth cells induced by platelet⁃derived growth factor⁃BB
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    摘要:

    目的:研究Rho激酶抑制剂ripasudil对血小板源性生长因子(platelet-derived growth factor,PDGF)-BB诱导人肺动脉平滑肌细胞(human pulmonary arterial smooth cells,HPASMCs)增殖和迁移的影响及其相关机制。方法:培养HPASMCs,随机分为control组、PDGF-BB组、PDGF-BB+ripasudil组、ripasudil组。采用CCK-8法检测细胞活力;EdU掺入法检测HPASMCs增殖;Transwell实验检测HPASMCs迁移;Real-time PCR检测基质金属蛋白酶(matrix metalloproteinase,MMP)-2 mRNA表达;Western blot检测MMP-2蛋白表达以及肌球蛋白磷酸酶目标亚基1(myosin phosphatase target subunit 1,MYPT1)、细胞外调节蛋白激酶1/2(extracellar regulated protein kinases 1/2,ERK1/2)、p38激酶和蛋白激酶B(protein kinase B,PKB/Akt)的磷酸化。结果:与control组相比,ripasudil能显著抑制PDGF-BB诱导HPASMCs增殖及迁移(P<0.01),降低MMP-2 mRNA及蛋白的表达(P<0.05),下调MYPT1、ERK1/2、p38及Akt的磷酸化(P<0.05)。结论:Ripasudil抑制PDGF-BB诱导的HPASMCs增殖和迁移,可能与下调MYPT1、ERK1/2、p38及Akt的磷酸化有关。Ripasudil可能是治疗肺动脉高压的潜在药物。

    Abstract:

    Objective:To investigate the effects of ripasudil on proliferation and migration of human pulmonary arterial smooth cells(HPASMCs)induced by platelet-derived growth factor(PDGF)-BB and the mechanisms underlying. Methods:Cultured HPASMCs were divided into four groups:the control group,the PDGF-BB-treated group,the PDGF-BB and ripasudil-treated group and the ripasudil-treated group. CCK-8 was applied to investigate cell viability and EdU assay was used to evaluate the proliferation of HPASMCs. Transwell assay was employed to examine cell migration. The expression of matrix metalloproteinase-2(MMP-2)was determined by real-time PCR and Western blot. The levels of phosphorylated myosin phosphatase target subunit 1(MYPT1),extracellar regulated protein kinases 1/2(ERK1/2),p38,and protein kinase B(PKB/Akt)were detected by Western blot. Results:Compared with the control group,ripasudil blocked the proliferation and migration of HPASMCs challenged by PDGF-BB(P<0.01). Ripasudil suppressed PDGF-BB-induced upregulation of MMP-2(P<0.05). Moreover,ripasudil inhibited PDGF-BB-induced phosphorylation of MYPT1,ERK1/2,p38,and Akt(P<0.05). Conclusion:Ripasudil significantly inhibited PDGF-BB-induced proliferation and migration of HPASMCs,which might be attributed to the inhibition of MYPT1,ERK1/2,p38 and Akt. Ripasudil,a Rho kinase inhibitor,might be a potential therapeutic option in PAH.

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孙德芳,孔 辉,解卫平,王 虹. Ripasudil对PDGF⁃BB诱导人肺动脉平滑肌细胞增殖和迁移的影响[J].南京医科大学学报(自然科学版),2018,(7):878-884

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  • 收稿日期:2018-03-29
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  • 在线发布日期: 2018-07-20
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