CAJ | 학술논문

目的评价高迁移率族蛋白B1(HMGB1)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× 105个/ml)后接种到96孔板(100 μl/孔,2× 105个/ml)、6孔板(1ml/孔,2× 106个/ml)和改良24孔Boyden趋化小室(100μg/孔,5×103个/ml),采用随机数字表法,将其分为5组:对照组(C组)和不同浓度HMGB1组(H1组～H4组),分别在DMEM和含HMGB1 1、10、100、1000 ng/ml的DMEM培养液孵育.孵育24和48 h时,采用MTT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,H3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)；H3组和H4组间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48 h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.
목적 평개고천이솔족단백B1(HMGB1)대체외배양인폐동맥혈관평활기세포(hPASMC)증식、천이화조망적영향.방법 체외배양hPASMC,조정세포밀도(2× 105개/ml)후접충도96공판(100 μl/공,2× 105개/ml)、6공판(1ml/공,2× 106개/ml)화개량24공Boyden추화소실(100μg/공,5×103개/ml),채용수궤수자표법,장기분위5조:대조조(C조)화불동농도HMGB1조(H1조～H4조),분별재DMEM화함HMGB1 1、10、100、1000 ng/ml적DMEM배양액부육.부육24화48 h시,채용MTT법검측세포증식솔,Boyden소실법검측투막세포수,TUNEL법검측hPASMC조망정황.결과 여C조비교,H1조～H4조세포증식솔승고,투막세포수증다(P＜ 0.05)；여H1조비교,H2조～H4조세포증식솔승고,H3조화H4조투막세포수증다(P＜0.05)；여H2조비교,H3조화H4조세포증식솔승고,투막세포수증다(P＜ 0.05)；H3조화H4조간각지표비교차이무통계학의의(P＞0.05)；여부육24h시비교,각조부육48 h시세포증식솔승고(P＜0.05).각조세포조망솔비교차이무통계학의의(P＞ 0.05).결론 HMGB1가촉진hPASMC적증식화투막천이,가능삼여폐손상폐혈관중구적발생.
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 (100 μl/well) with the density of 2 × 105/ml,in 6-well plate (1 ml/well) with the density of 2 × 106/ml or in modified 24-well Boyden chamber (100 μl/well) with the density of 5 × 103/ml and randomly divided into 5 groups (n =26 each):control group (group C) and 4 different concentrations of HMGB1 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 MTT,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.