丹参酮通过调控巨噬细胞分化保护OGD/R诱导的心肌细胞凋亡的作用

Tanshinone protects cardiomyocytes from OGD/R-induced apoptosis by regulating macrophage differentiation

目的探讨丹参酮通过调控巨噬细胞分化保护OGD/R诱导的心肌细胞凋亡及相关机制。方法选用THP-1单核细胞株经佛波醇12-十四酸酯13-乙酸酯(PMA)诱导为贴壁的巨噬细胞后,分为丹参酮低剂量组(1μg/ml)、丹参酮中剂量组(2μg/ml)和丹参酮高剂量组(4μg/ml),荧光定量PCR检测M1型和M2型巨噬细胞活化相关基因表达。采用巨噬细胞和心肌细胞共培养的方式探索巨噬细胞的分化对OGD/R诱导的心肌细胞凋亡的影响,Tunel染色检测心肌细胞凋亡情况,Western blot检测心肌细胞凋亡相关蛋白Bax、Bcl2和Caspase3表达以及NF-κB信号通路相关蛋白p-IKB、IKB、p-P65和P65表达。使用NF-κB信号通路激动剂探索巨噬细胞的分化对OGD/R诱导的心肌细胞凋亡影响的相关机制,进行上述相同的检测。结果丹参酮低、中和高剂量都可抑制M1型巨噬细胞分化相关基因表达,促进M2型巨噬细胞分化相关基因(P<0.05),其中丹参酮中剂量效果最佳(P<0.01)。相关机制研究发现,丹参酮可通过调控巨噬细胞分化过程抑制OGD/R诱导的心肌细胞凋亡(P<0.05)以及凋亡相关蛋白Bax/Bcl2和Cleaved-caspase3/Caspase3表达(P<0.05)。相关信号通路研究发现,M2型巨噬细胞活化相关因子主要通过影响NF-κB信号通路相关蛋白p-IKB和p-P65的表达抑制OGD/R诱导的心肌细胞凋亡(P<0.05)。使用NF-κB信号通路激动剂研究结果显示,激活NF-κB信号通路能够阻碍丹参酮通过调控巨噬细胞分化抑制OGD/R诱导的心肌细胞凋亡过程。结论丹参酮通过调控巨噬细胞分化,影响NF-κB信号通路,进而抑制OGD/R诱导的心肌细胞凋亡。

To investigate the protective effect of tanshinone on cardiomyocyte from OGD/R-induced apoptosis by regulating macrophage differentiation and to explore the related mechanisms.THP-1 mononuclear cell lines were induced into adherent macrophages by phorbo alcohol 12-tetradecanoate 13-acetate(PMA)and divided into tanshinone low-dose group(1μg/ml),tanshinone medium-dose group(3μg/ml)and tanshinone high-dose group(5μg/ml).The expression of M1/M2-type cell activation-related genes were detected by fluorescence quantitative PCR.Macrophages and cardiomyocytes were co-cultured to explore the effect of macrophage differentiation on cardiomyocyte apoptosis induced by OGD/R.Tunel staining was used to detect myocardial cell apoptosis;Western blot was used to detect the expression of apoptosis-related proteins Bax,Bcl2 and Caspase3,as well as NF-κB signaling pathway related proteins p-IKB,IKB,p-P65 and P65.NF-κB signaling pathway agonists were used to explore the related mechanisms of macrophage differentiation affecting OGD/R induced cardiomyocyte apoptosis.Data showed that low,medium and high doses of tanshinone inhibited the expression of M1-type macrophage differentiation-related genes and promoted the expression of M2-type macrophage differentiation-related genes(P<0.05),among which tanshinone of middle dose showed the best effecacy(P<0.01).Tanshinone inhibited OGD/R-induced apoptosis of cardiomyocytes by regulating macrophage differentiation(P<0.05)and apoptosis-related proteins Bax/Bcl2 and cleaved-caspase3/Caspase3(P<0.05).It was found that M2-type macrophage activation-related factors inhibited OGD/R-induced apoptosis of cardiocytes by affecting the expression of NF-κB signaling pathway-related proteins p-IKB and p-P65(P<0.05).Activation of NF-κB signaling pathway can prevent tanshinone from inhibiting OGD/R induced cardiomyocyte apoptosis by regulating macrophage differentiation(P<0.05).Taken together,tanshinone can inhibit OGD/Rinduced cardiomyocyte apoptosis by regulating macrophage differentiation and influencing NF-κB signaling pathway.