CAJ | 학술논문

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。
목적：탐토치료병태두방결종합정적통양활혈방함약혈청대모의결혈재관주손상토두방결세포조망화세포골가단백Vinculin적영향。방법취신생유토두방결세포，이결양결당모의결혈，이회복양화당적공응모의재관주조성두방결세포손상모형。설정상대조조、모형조、공백혈청조、함약혈청고、중、저제량조，운용매표의、류식세포분석의、격광공취초현미경관찰각조두방결세포활성、조망급세포골가단백Vinculin형태적변화。결과모형조활세포량교정상조명현감소(P<0.01)；조망명현증다(P<0.01)；공백혈청조여모형조무명현차이。세포골가단백Vinculin렬해명현。함약혈청고、중、저제량조활세포량명현고우모형조(P<0.05)，조망명현감소(P<0.01)， Vinculin결구교모형조완정，형광강도명현고우모형조。결론통양활혈방함약혈청능억제모의결혈재관주인기적두방결세포손상；통양활혈방치료병태두방결종합정적궤제가능여보호두방결세포활성、대항세포조망、보호세포골가단백Vinculin형태결구유관。
Objective To observe the infulence of Tongyang Huoxue Formula drug serum on apoptosis and Vinculin expression of sinoatrial node cells of rabbit with ischemia reperfusion. Methods Sinoatrial node cells of newborn rabbits were separated and used to establish the sinoatrial node cells damage models by using methods of oxygen-glucose deprivation instead of ischemia and oxygen-glucose supply instead of reperfusion. The research included control group, model 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<0. 05), and cell apoptosis increased obviously (P<0. 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 reperfusion, and the mechanism might be related with protective effect in cell viability and cytoskeletal protein form and anti-apoptosis effect.