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/m L)、TGF-β阳性对照组(1 ng/m L),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/m L)刺激HASMC后,细胞迁移能力显著高于对照组(P〈0.01);HMGB1(500 ng/m L)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。
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 HMGBl-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 HMGB 1 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.