结缔组织生长因子PD98059和LY294002在调节大鼠肺动脉平滑肌细胞增殖迁移中的作用

doi:10.3969/j.issn.1006-6233.2020.02.015

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摘要 目的:探讨大鼠肺动脉平滑肌细胞(PASMC)在结缔组织生长因子(CTGF)、PD98059和LY294002干预下的增殖迁移。方法:分离培养大鼠PASMC,并分为空白组、CTGF组、CP组、CL组和CPL组,WST-1检测CTGF、PD98059和LY294002对细胞增殖的影响,Transwell、细胞划痕和细胞骨架检测其对细胞迁移的影响。结果:与空白组比较,CTGF(50ng/mL)刺激PASMC48、72、96和120h后WST-1示细胞增殖水平升高,6h、12h后Transwell示穿膜细胞数量增加,12h、24h后细胞划痕示划痕宽度减小,细胞骨架模糊,差异有统计学意义(P<0.05)。与CTGF组比较,经PD98059(20μmoL/L)和/或LY294002(10μmoL/L)干预24、48、72、96和120h后WST-1示细胞增殖水平下降;6h、12h后Transwell示穿膜细胞数量减少,12h、24h后细胞划痕示划痕宽度增加,细胞骨架清晰,差异有统计学意义(P<0.05)。结论:CTGF可刺激PASMC增殖迁移并可能是促进肺动脉高压(PAH)肺血管重构因素之一,PASMC增殖迁移可被PD98059和/或LY294002抑制,提示它们所阻断的ERK1/2和PI3K通路可能参与了CTGF刺激下的PAH肺血管重构。

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关键词 ：结缔组织生长因子, 大鼠肺动脉平滑肌细胞, 增殖迁移

Abstract：Objective: To explored the proliferation and migration of (PASMC) in rat pulmonary artery smooth muscle cells under the intervention of connective tissue growth factor (CTGF), PD98059 and LY294002. Method: PASMC was isolated and cultured, and then were divided into blank group, CTGF group, CP Group, CL group and CPL group. The effects of CTGF, PD98059 and LY294002 on cell proliferation were detected by WST-1, and the effects on cell migration were detected by Transwell, cell scratch and cytoskeleton. Results: Compared with the blank group, the proliferation level of WST-1 cells increased after CTGF (50NG/ml) stimulation of pasmc48, 72, 96 and 120 hours, the number of transmembrane cells increased after 6h and 12h, the scratch width of cells decreased after 12h and 24h, and the cytoskeleton was fuzzy, the difference was statistically significant (P<0.05). After PD98059 (20 μ mol / L) and / or LY294002 (10 μ moL/L) intervention for 24, 48, 72, 96 and 120 hours, WST-1 showed a decrease in cell proliferation; after 6 and 12 hours, Transwell showed a decrease in the number of transmembrane cells, and after 12 and 24 hours, the scratch width and cytoskeleton of the cells were increased, the difference was statistically significant (P<0.05). Conclusion: CTGF can stimulate the proliferation and migration of PASMC and may be one of the factors to promote the pulmonary vascular remodeling of pulmonary hypertension (PAH). PASMC proliferation and migration can be inhibited by PD98059 and / or LY294002, suggesting that ERK1 / 2 and PI3K pathways blocked by CTGF may participate in the pulmonary vascular remodeling of PAH stimulated by CTGF.

Key words： Connective tissue growth factor Pulmonary artery smooth muscle cells of rat Proliferation migration

基金资助:四川省卫生厅科研基金资助,(编号:110531)

通讯作者: 刘斌

引用本文:

胡煜, 周孟君, 李刚, 刘斌, 余静. 结缔组织生长因子PD98059和LY294002在调节大鼠肺动脉平滑肌细胞增殖迁移中的作用[J]. 河北医学, 2020, 26(2): 235-239.
HU Yu, ZHOU Mengjun, et al. Role of Connective Tissue Growth Factor PD98059 and LY294002 in Regulating the Proliferation and Migration of Pulmonary Artery Smooth Muscle Cells of Rat. HeBei Med, 2020, 26(2): 235-239.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2020.02.015 或 http://www.hbyxzzs.cn/CN/Y2020/V26/I2/235