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作 者:钱睿哲[1] 张国平[1] 金惠铭[1] 王文健[2] 乐飞[1] 史连国[1] 曲晓义[1]
机构地区:[1]复旦大学上海医学院生理与病理生理学系,上海200032 [2]复旦大学附属华山医院中西医结合研究所,上海200040
出 处:《中国病理生理杂志》2005年第11期2086-2090,共5页Chinese Journal of Pathophysiology
基 金:SupportedpartiallybythekeysubjectofdevelopmentfoundationfromtheNational10thfiveyearplan,211project,Shanghaiscienceandtechnologyfoundation(No.014319321)andUnileverResearchandDevelopmentfoundation.
摘 要: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.
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