摘要
目的 :研究苦参碱 (matrine,Mat)诱导血管紧张素Ⅱ (AngⅡ )作用的小鼠心肌成纤维细胞 (cardiacfi broblast,CFB)凋亡及相关调控蛋白表达的影响。方法 :不同浓度苦参碱作用于体外培养AngⅡ作用的小鼠心肌成纤维细胞 ,利用倒置显微镜、PI/Hoechst 33342双染结合荧光显微镜技术观察细胞形态 ;流式细胞术检测其凋亡率、细胞周期及凋亡相关基因Bcl 2、Bax蛋白表达的改变 ;采用WesternBlotting检测Caspase 3蛋白的表达。 结果 :不同浓度Mat处理AngⅡ作用的小鼠心肌成纤维细胞 2 4h ,可见到染色质浓缩、边集、碎裂 ;凋亡率随浓度的升高而升高 ;细胞周期分析显示G1升高 ,S期下降 ,G2 /M期无明显变化 ;,Bcl 2的表达量减少 ;Bax的表达量增加且呈剂量效应 ;WesternBlotting检测显示Caspase 3活化。结论 :Mat诱导AngⅡ作用的小鼠心肌成纤维细胞凋亡且呈剂量效应 ,其机制可能与降低Bcl 2、升高Bax、增强Caspas 3活性有关。
Objective: To investigate the apoptosis effects of matrine on angiotensinⅡ-induced mouse cardiac fibroblasts. Methods: Cardiac fibroblasts of mice were cultured and cell count,morphology of apoptosis cells were studied by fluorescence microscope with PI/Hoechst 33342 double staining. Cell cycle phases,apoptosis ratio of angiotensinⅡ-induced mouse cardiac fibroblasts by matrine treatment and expression of the apoptosis-related gene Bcl-2 and Bax were observed by FCM. Chemiluminescence Western blotting analysis of total cell extracts was used to detect Caspase-3 expression.Results: Treatment of angiotensinⅡ-induced mouse cardiac fibroblasts with concentrations of 1～4 mmol·L -1 matrine for 24 hours revealed that the apoptosis ratio was increased gradually as the concentration was increased( P <0.05). PI/Hoechst 33342 double staining could distinguish apoptosis cells from normal cells. Typical apoptosis features of cells were observed under fluorescence microscope in 3.0 mmol·L -1 matrine + angiotensinⅡgroup and the features were more prominent as the concentration was increased. Matrine also affected cell cycle obviously,mainly causing to increase the number of cells in G_1 phase ( P <0.05) and to decrease in S phase ( P <0.05). Expression of the apoptosis-suppression gene Bcl-2 was reduced ( P < 0.05 ),whiled expression of the apoptosis-induction gene Bax was enhanced,and the effects was dose-dependent ( P <0.05). At the same time activity of Caspase-3 was induced. Conclusion: The effects of matrine on apoptosis of angiotensinⅡ-induced mouse cardiac fibroblasts is dose-dependent and it might be mediated by the changes in the expression of the apoptosis-related gene Bcl-2,Bax and Caspase-3.
出处
《武汉大学学报（医学版）》
CAS
2004年第4期375-379,T002,共6页
Medical Journal of Wuhan University
