穿心莲酸对胆固醇结晶诱导的THP-1源性巨噬细胞损伤的影响

Effect of andrographolic acid on THP-1-derived macrophage damage induced by cholesterol crystals

目的探讨穿心莲酸对胆固醇结晶所致THP-1源性巨噬细胞损伤的干预保护作用及其作用机理。方法用10 nmol/L佛波酯(PMA)诱导THP-1单核细胞分化为巨噬细胞,加入胆固醇结晶孵育的同时给药组给予相应的药物干预24 h,用显微镜观察细胞状态,收集细胞并提取总RNA,RT-qPCR检测相关基因mRNA的表达水平。结果与正常巨噬细胞相比,模型组巨噬细胞形态变化明显,细胞出现不规则轮廓、膨胀。RT-qPCR检测发现,模型组细胞自噬相关基因P62、beclin-1、LC3和凋亡相关基因P53、Bcl-2、Bax、Caspase-8、Caspase-9、BIM等明显升高(P<0.05);而与模型组相比,穿心莲酸低剂量组显著下调自噬相关基因和凋亡相关基因的mRNA表达水平(P<0.05),瑞舒伐他汀组与穿心莲酸高剂量组能下调部分基因的mRNA表达水平(P<0.05)。结论穿心莲酸通过减少自噬的发生使细胞免受胆固醇结晶所致凋亡及损伤,从而起到对巨噬细胞的保护修复作用。

Objective To study the protective effect and mechanism of andrographolic acid on THP-1-derived macrophage damage induced by cholesterol crystals.Methods THP-1monocytes were differentiated into macrophages by treatment with10nmol/L PMA.The macrophages were treated with agents and/or cholesterol crystals for24h in experiment groups.The cell morphology was observed with the microscope.The expression of related gene mRNA was detected by RT-qPCR.Results The macrophages in the model group showed significant morphological changes including irregular membrane and cell foaming.The expressions of autophagy-related genes P62,beclin-1and LC3and apoptosis-related genes P53,Bcl-2,Bax,Caspase-8,Caspase-9and BIM were significantly increased in model group when compared to the normal group(P<0.05).Compared with the model group,the expressions of autophagy and apoptosis related genes were significantly downregulated in the low dosage of andrographolic acid group(P<0.05),but the changes in the high dosage of andrographolic acid and rosuvastatin groups were not significant.Conclusion Andrographolic acid protects THP-1-derived macrophages against cholesterol crystal-induced cell apoptosis and damage by reduction of cell autophagy.