高迁移率族蛋白B1对体外人肺动脉血管平滑肌细胞增殖、迁移和凋亡的影响

Effects of high mobility group protein box 1 on proliferation, migration and apoptosis of human pulmonary artery smooth muscle cells in vitro

目的评价高迁移率族蛋白B1（HMGBl）对体外培养人肺动脉血管平滑肌细胞（hPASMC）增殖、迁移和凋亡的影响。方法体外培养hPASMC，调整细胞密度（2×10^5个／m1）后接种到96孔板（100μl／孔，2×10^5个／m1）、6孔板（1ml／孔，2×10^5个／ml）和改良24孔Boyden趋化小室（100μg/孔，5×10^5个／m1），采用随机数字表法，将其分为5组：对照组（C组）和不同浓度HMGBI组（H1组～H4组），分别在DMEM和含HMGB 11、10、100、1000ng／ml的DMEM培养液孵育。孵育24和48h时，采用MTY法检测细胞增殖率。Boyden小室法检测透膜细胞数，TUNEL法检测hPASMC凋亡情况。结果与C组比较，H1组～H4组细胞增殖率升高，透膜细胞数增多（P〈0．05）；与H1组比较，H2组-H4组细胞增殖率升高，地组和H4组透膜细胞数增多（P〈0．05）；与H2组比较，H3组和H4组细胞增殖率升高，透膜细胞数增多（P〈0．05）；H，组和Hd组问各指标比较差异无统计学意义（P〉0．05）；与孵育24h时比较，各组孵育48h时细胞增殖率升高（P〈0．05）。各组细胞凋亡率比较差异无统计学意义（P〉0．05）。结论HMGB1可促进hPASMC的增殖和透膜迁移，可能参与肺损伤肺血管重构的发生。

Objective To evaluate the effects of high mobility group protein box 1 （HMGB1） on the proliferation, migration and apoptosis of human pulmonary artery smooth muscle cells （hPASMCs） in vitro. Methods The hPASMCs were seeded in 96-well plates in DMEM liquid culture medium （100μl/well） withthe density of 2 × 10^5/ml, in 6-well plate （1 ml/well） with the density of 2 × 10^6/ml or in modified 24-well Boyden chamber （100 μl/well） with the density of 5 × 10^3/ml and randomly divided into 5 groups （n = 26 each）： control group （group C） and 4 different concentrations of HMGBI groups （groups H1 -H4 ）. The cells were cultured in DMEM liquid culture medium in group C. The cells were cultured in DMEM liquid culture medium containing HMGB1 1, 10, 100 and 1000 ng/ml in groups H1-H4, respectively. At 24 and 48 h of incubation, the proliferation, migration and apoptosis of hPASMCs were detected by MTY, modified Boyden chamber assay, and TUNEL, respectively. Results Compared with group C, the proliferation and migration of hPASMCs were significantly increased in groups H1-H4 （ P 〈 0.05）. The proliferation of hPASMCs was significantly higher in groups H2 -H4 , and the migration of hPASMCs was higher in groups H3 and H4 than in group H1, and in groups H3 and H4 than in group H2 （ P 〈 0.05）. There was no significant difference in the parameters mentioned above between groups H3 and H4 （ P 〉 0.05）. The proliferation of hPASMCs was significantly higher at 48 h of incubation than at 24 h of incubation （ P 〈 0.05）. There was no significant difference in the apoptotic rate between the five groups （ P 〉 0.05 ）. Conclusion HMGB1 can induce the proliferation and migration of hPASMCs in vitro, which may be involved in pulmonary vascular remodeling during acute lung injury.