高迁移率族蛋白B1对人气道平滑肌细胞增殖和迁移及Akt磷酸化的影响
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国家自然科学基金(81370133?81170018);江苏省医学重点人才项目(RC2011066);江苏省高校自然科学研究项目(11KJB320008);江苏省临床医学科技专项(BL2012012);江苏省社会发展重点研发专项(BE2015651);江苏省卫生计生委预防医学科研课题(Y2015026);江苏省中医药科技项目(YB2015110);苏州市科技计划项目(SYS201402)


Effects of high-mobility group box protein 1 (HMGB1) on the proliferation, migration and phosphorylation of Akt in human airway smooth muscle cells
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    摘要:

    目的:探讨高迁移率族蛋白B1(high morbility group protein box 1,HMGB1)刺激对人气道平滑肌细胞(human airway smooth muscle cells,HASMC)增殖和迁移的影响及其机制。方法:体外培养HASMC,将纯化的第3~8代细胞随机分为对照组-HMGB1组(125-250-500和1 000 ng/mL)-TGF-β阳性对照组(1 ng/mL),CCK-8法测定HASMC增殖能力,Transwell小室观察HMGB1对 HASMC迁移的影响,Western blot检测HASMC的Akt磷酸化。结果:与对照组相比,HMGB1呈浓度依赖性刺激HASMC增殖(P < 0.05);HMGB1(500 ng/mL)刺激HASMC后,细胞迁移能力显著高于对照组(P < 0.01);HMGB1(500 ng/mL)组HASMC的Akt磷酸化水平较对照组显著增高(P < 0.05)。结论:HMGB1促进HASMC增殖和迁移,可能是通过调节PI3K/Akt通路参与气道重塑,为今后治疗哮喘提供了新思路。

    Abstract:

    Objective:To investigate the effects and underlying mechanism of high mobility group protein box 1(HMGB1) on the proliferation and migration of human airway smooth muscle cells(HASMCs). Methods:Normal HASMC between passages 3 and 8 were randomly divided into the control group, HMGB1 group with different concentrations(125, 250, 500, and 1 000 ng/mL), and TGF-β(1 ng/mL) group as a positive control, respectively. The HMGB1-stimulated proliferation of HASMCs was evaluated by cell counting of CCK-8. The number of migrated cells was performed using Transwell migration assay. Western blotting assay was performed to detect the phosphorylation of Akt following HMGB1 stimulation. Result: The proliferation of HASMCs treated with HMGB1 was significantly increased in a dose-dependent manner than those of untreated control group (P < 0.05). Furthermore, the number of migrated cells was significantly increased after HMGB1 activation compared to the control group(P < 0.01). The phosphorylation level of Akt in the HMGB1 group(500 ng/mL) was significantly increased compared with the control group(P < 0.05). Conclusion:Our data suggest that HMGB1 contributes to airway remodeling by stimulating the proliferation and migration of HASMCs through regulating phosphorylation of PI3K/Akt pathway, which provides a promising target for future therapy of asthma.

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柏芳芳,陈 子,韩 露,闫 兆,王 楠,解卫平,黄 茂,周林福.高迁移率族蛋白B1对人气道平滑肌细胞增殖和迁移及Akt磷酸化的影响[J].南京医科大学学报(自然科学版),2016,(7):794-797

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  • 收稿日期:2015-12-01
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  • 在线发布日期: 2016-07-15
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