强心复脉颗粒药物血清对模拟缺血再灌注致钙超载窦房结细胞的影响

Effect of Qiangxin Fumai Granule Contained Serum on Sinoatrial Node Cells during Ca^(2+) Overloading Induced by Simulated Ischemia/Reperfusion

目的探讨治疗病态窦房结综合征的强心复脉颗粒药物血清对模拟缺血再灌注致钙超载窦房结细胞的影响。方法取新生乳鼠窦房结细胞,以缺氧缺糖模拟缺血,以恢复氧和糖供应模拟再灌注造成窦房结细胞的钙超载模型。设正常对照、模型对照、阿托品药物血清、强心复脉药物血清高、低剂量组,给药组用含相应药物血清的DMEM培养基在有氧环境中预先培养30 min后,再进行模拟缺血再灌注。运用倒置显微镜、荧光分光光度计观察各组窦房结细胞形态和钙离子浓度的变化。结果模型对照组大部分细胞出现肿胀变形,甚至细胞脱落;HE染色后观察,胞质内出现较多微小空泡,胞核核膜有皱褶,胞膜局部破损。强心复脉药物血清高、低剂量组细胞贴壁生长良好,胞膜、核膜基本平滑完整,细胞大小形态正常。正常窦房结细胞[Ca^(2+)]i值较低;模拟缺血再灌注后显著增高;强心复脉药物血清高、低剂量组窦房结细胞[Ca^(2+)]i明显降低(P<0.01),而阿托品药物血清组无明显降低。结论强心复脉药物血清可抑制模拟缺血再灌注引起的细胞损伤。强心复脉颗粒治疗病态窦房结综合征的机制可能与其能降低钙离子浓度、保护窦房结细胞损伤有关。

Objective To explore the effect of Qiangxin Fumai Granule （QFG, a Chinese herbal preparation for treatment of sick sinus syndrome） contained serum （QFG-S） on sinoatrial node cells during Ca2＋ overloading induced by simulated isehemia/reperfusion. Methods Model Ca2＋ overloading ceils were established on sinoatrial node cells from newborn rats, with deprivation of oxygen and glucose to simulate ischemia and with restoration of them to simulate reperfusion. Cells were divided into 5 groups, those in the normal and model control groups were modeled directly and those in the treated groups were pre-cuhured with DMEM containing respective medicines in aerobic environment for 30 min before ischemia/reperfusion simulation. Cell Ca2＋ concentration and morphology were observed by invert microscope and fluorescence speetrophotometer. Results Most cells in the model control group revealed cell edema and deformation, even abscission. By HE staining, many minimal vacuole appeared in cytoplasm, with crumpled nuclear membrane, paltially damaged. While cells in the QFG-S treated group, either high-dose or low-dose, were attached grew well, with basically smooth and complete membrane and nuclear membrane, normal in size and shape. The intracellular Ca2＋ concentration raised significantly after modeling, but it was much lower in the QFG-S treated group than in the model control group （P 〈 0.01 ） and it was not changed obviously in the atropine treated group. Conclusion QFG-S could diminish the injury of cell induced by simulated ischemia/reperfusion, the acting mechanism for treatment of sick sinus syndrome might be related to its effect in relieving Ca2＋ overloading and thus protecting cells from injury.