大花红天素对脑微血管内皮细胞凋亡的双相调节作用及机制(英文)

Dual-direction effect of crenulatin on apoptosis of cerebral microvascular endothelial cells and it's mechanism

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。

AIM： To study the effect and the mechanism of crenulatin, an effective constituent of Chinese traditional medicine, on apoptosis of cerebral microvascular endothelial cells. METHODS： The following terminal concentrations of crenulatin were used in the study： 25 mg/L and 100 mg/L. Apoptosis of mouse cerebral microvascular endothelial cells （bEnd.3 cell line） was evaluated by flow cytometer, immunocytochemical assay （Fas, Bcl- 2） and Western blotting （caspase- 3） after culture for 24 h. RESULTS： Compared with control group, apoptosis of bEnd.3 cells in 25 mg/L group was significantly inhibited （ P 〈 0.05）, but apoptosis in the 100 mg/L group was significantly increased （ P 〈 0.05）. In apoptosis inhibited group, the Fas immunocytochemical staining was weaker, the positive cells were significantly decreased （ P 〈 0.05） and caspase - 3 expression was decreased compared with control group; however, the Bcl - 2 staining was stronger and the positive cells were significantly increased （ P 〈 0.05）. On the other hand, in apoptosis increased group （ 100 mg/L group）, the changes were just opposite. CON- CLUSIONS： The effect of crenulatin on apoptosis of mouse cerebral microvascular endothelial cells possesses a dual - direction change, inhibitive effect in 25 mg/L and stimulative effect in 100 mg/L group, respectively. The mechanism is related to the alterations of Fas/Bcl - 2 expression and caspase - 3 activity.