摘要
目的探讨低氧对血管平滑肌细胞分泌高迁移率族蛋白B1(HMGB1)的影响,以及尼莫地平对其干预作用。方法体外培养大鼠血管平滑肌细胞,采用Kuwabara等并加以改进的方法对平滑肌细胞进行低氧培养,在低氧处理后(1h、4h、12h、24h和48h),使用Westernblot检测培养液上清中HMGB1的表达水平。再按同样方法加入尼莫地平预处理,分为对照组、单纯低氧组、低氧+尼莫地平组(加入100mmol/L尼莫地平),低氧培养24h后检测上清HMGB1水平。台盼蓝排斥法检测48h以内细胞的存活率。结果与正常对照组相比,培养液上清中HMGB1在低氧各组表达均明显增加(P<0.05)。低氧组、低氧+尼莫地平组HMGB1水平明显高于对照组,尼莫地平预处理组较单纯缺氧组要低(P<0.05)。台盼蓝排斥法检测低氧48h以内对细胞生存率影响较小。结论低氧可诱导体外培养的血管平滑肌细胞分泌表达HMGB1,在24h内随着缺氧时间的延长,其表达增加。加入尼莫地平预处理可降低HMGB1的表达,故提示尼莫地平可能通过部分降低HMGB1水平从而起到脑保护作用。
Objective To observe the effects of hypoxia on the expression of HMGB1 in cultured VSMCs. Methods The cultured VSMCs of rats were divided into six groups and they were cultured in normal condition, and under hypoxic condition for 1, 4, 12, 24 and 48 hours. The levels of HMGB1 in the supernatant were measured by Western blot. Subsequently, we administrated nimodipine to pretreatment. The cells were divided into three groups： normal control group,hypoxic group and nimodipine-pretreated hypoxic group. HMGB1 were measured after 24 hours＇ hypoxic culture. The cell viability within 48h was examined by trypan blue exclusion test. Results Compared with normal control, the expression of HMGB1 in the hypoxic cultured supernatant significantly increased（P0.05）. But the HMGB1 level in nimodipine-pretreated hypoxic group was lower than in the hypoxic group（P0.05）.Within 48h, no differences in cell viability were found compared with control group by Trypan blue exclusion test. Conclusion Hypoxia can significantly induce the expression of HMGB1 in cultured VSMCs and the expression increase gradually within 24h. Nimodipine can significantly decrease the expression of HMGB1 in hypoxic condition. We presume that the mechanisms of brain protection by nimodipine may be mediated in part by decreasing the HMGB1 level.
出处
《脑与神经疾病杂志》
2010年第6期429-432,共4页
Journal of Brain and Nervous Diseases
