通阳活血方含药血清对兔缺血再灌注窦房结细胞凋亡和细胞骨架蛋白Vinculin的影响  

Infulence of Tongyang Huoxue Formula drug serum on apoptosis and Vinculin expression of sinoatrial node cells of rabbit with inchemia reperfusion

作　　者：刘如秀[1] 彭杰[1] 刘宇[1] 

机构地区：[1]中国中医科学院广安门医院心内科,北京100053

出　　处：《环球中医药》2015年第10期1182-1186,共5页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81173447)

摘　　要：目的探讨治疗病态窦房结综合征的通阳活血方含药血清对模拟缺血再灌注损伤兔窦房结细胞凋亡和细胞骨架蛋白Vinculin的影响。方法取新生乳兔窦房结细胞,以缺氧缺糖模拟缺血,以恢复氧和糖的供应模拟再灌注造成窦房结细胞损伤模型。设正常对照组、模型组、空白血清组、含药血清高、中、低剂量组,运用酶标仪、流式细胞分析仪、激光共聚焦显微镜观察各组窦房结细胞活性、凋亡及细胞骨架蛋白Vinculin形态的变化。结果模型组活细胞量较正常组明显减少(P<0.01);凋亡明显增多(P<0.01);空白血清组与模型组无明显差异。细胞骨架蛋白Vinculin裂解明显。含药血清高、中、低剂量组活细胞量明显高于模型组(P<0.05),凋亡明显减少(P<0.01),Vinculin结构较模型组完整,荧光强度明显高于模型组。结论通阳活血方含药血清能抑制模拟缺血再灌注引起的窦房结细胞损伤;通阳活血方治疗病态窦房结综合征的机制可能与保护窦房结细胞活性、对抗细胞凋亡、保护细胞骨架蛋白Vinculin形态结构有关。Objective To observe the infulence of Tongyang Huoxue Formula drug serum on apoptosis and Vinculin expression of sinoatrial node cells of rabbit with ischemia repeffusion. Methods Sinoatrial node cells of newborn rabbits were separated and used to establish the sinoatrial node ceils damage models by using methods of oxygen-glucose deprivation instead of ischemia and oxygen-glucose supply instead of reperfusion. The research included control group, nlodel group, blank serum group, and high, middle and low dosage of Tongyang Huoxue Formula drug serum group. Microplate reader, flow cytometer and laser scanning confocal microscope were used to observe the activity, apoptosis and Vinculin expression of sinoatrial node cells. Results Compared with control group, the living cells number of model group were lessen significantly （P〈O. 05 ）, and cell apoptosis increased obviously （P〈O. 01 ）. The difference between blank serum group and model group was not significant. The cleavage of cytoskeletal protein Vinculin was obviously. Compared with model group, the living cells number of high, middle and low dosage of drug serum groups were higher （ P〈0.05 ）, cell apoptosis decreased significantly （ P〈0. 01 ）, and the structure of Vinculin was more integrity and fluorescence intensity was stronger. Conclusions Tongyang Huoxue Formula drug serum could inhibit the sinoatrial node cells damage induced by simulated ischemia repeffusion, and the mechanism might be related with protective effect in ceil viability and cytoskeletal protein form and anti-apoptosis effect.

关 键 词：通阳活血方 含药血清 窦房结细胞 缺血再灌注 细胞凋亡 细胞骨架 

分 类 号：R285.5[医药卫生—中药学]