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目的研究中药红景天的有效成分-大花红天素对小鼠脑微血管内皮细胞凋亡的作用及机制。方法观察不同剂量的大花红天素(25mg/L,100mg/L)对体外培养的小鼠脑微血管内皮细胞(bEnd.3株)凋亡的影响。细胞凋亡分别用流式细胞术、免疫细胞化学ABC法(Fas/Bcl-2)及Western blotting(caspase-3)测定。结果与对照组比,在bEnd.3细胞中含药浓度25mg/L组凋亡明显抑制(P<0.05),而100mg/L组凋亡明显加剧(P<0.05)。凋亡抑制组Fas免疫细胞化学染色比对照组弱,阳性细胞明显减少(P<0.05);而Bcl-2染色比对照组强,阳性细胞明显增加(P<0.05),caspase-3表达明显抑制(P<0.05)。凋亡加剧组中(100mg/L)变化相反。结论大花红天素对bEnd.3细胞凋亡有双相调节作用,其机制与Fas/Bcl-2蛋白表达及caspase-3激活的变化有关。
Objective To study the effect of rhodopsin, an effective component of Rhodiola crenulata, on the apoptosis of mouse brain microvascular endothelial cells and its mechanism. Methods The effects of different doses of macrophyllin (25mg/L, 100mg/L) on apoptosis of mouse brain microvascular endothelial cells (bEnd.3) cultured in vitro were observed. Apoptosis was determined by flow cytometry, immunocytochemistry ABC method (Fas/Bcl-2) and Western blotting (caspase-3). Results Compared with the control group, apoptosis was significantly inhibited in the bEnd.3 cells at a concentration of 25 mg/L (P<0.05), whereas apoptosis was significantly increased in the 100 mg/L group (P<0.05). Fas immunocytochemical staining in the apoptosis-inhibitory group was weaker than that in the control group, and the number of positive cells was significantly decreased (P<0.05). Bcl-2 staining was stronger than the control group, positive cells were significantly increased (P<0.05), and caspase-3 expression was significantly inhibited (P<0.05). The change in the aggravated apoptosis group (100 mg/L) was opposite. Conclusion Macrophyllin has a biphasic effect on the apoptosis of bEnd.3 cells, and its mechanism is related to the expression of Fas/Bcl-2 protein and the activation of caspase-3.