基于CAV1.2/CaM/CaMKⅡ通路探讨祛痰化瘀中药复治疗心房颤动的作用机制  

Based on CAV1.2/CaM/CaMKⅡPathway to Explore the Mechanism of the Treatment of Atrial Fibrillation with Traditional Chinese Medicine Complex for Removing Phlegm and Removing Blood Stasis

作　　者：吴启华 赵帅 郝颖[1] 宫丽鸿[2] Wu Qihua;Zhao Shuai;Hao Ying;Gong Lihong(The first Clinical College,Liaoning University of Traditional Chinese Medicine,Shenyang 110033,China;Affiliated hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China)

机构地区：[1]辽宁中医药大学第一临床学院,沈阳110033 [2]辽宁中医药大学附属医院,沈阳110032

出　　处：《世界科学技术-中医药现代化》2023年第11期3660-3667,共8页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：辽宁省教育厅高校科研基金(21601A1088):辽宁省特聘教授课题项目,负责人:宫丽鸿;辽宁省科学技术厅应用基础研究计划(2022JH2/101300086):中药复方治疗痰浊血瘀型房颤的疗效及机制研究,负责人:宫丽鸿

摘　　要：目的探讨祛痰化瘀中药复方治疗心房颤动的作用机制。方法将60只雄性SD大鼠随机分为空白组和造模组,连续7天尾静脉注射Ach(66μg·mL^(-1))-CaCl_(2)(10 mg·mL^(-1))建立大鼠AF模型。将造模成功的大鼠随机分为模型组、中药复方高、中、低剂量组和维拉帕米组,中药复方高、中、低剂量组给予12.38 mg·kg^(-1)·d^(-1)、6.18 mg·kg^(-1)·d^(-1)、3.10 mg·kg^(-1)·d^(-1)祛痰化瘀中药复方溶液灌胃、维拉帕米组给予维拉帕米溶液8.31 mg·kg^(-1)·d^(-1)灌胃,空白组、模型组给予等体积蒸馏水灌胃,期间仍持续尾静脉注射,连续14天。电生理记录仪测量大鼠Ⅱ导联房颤持续时间,透射电镜观察大鼠心房肌超微结构变化,RT-PCR法检测大鼠心房肌CAV1.2、CaM、CaMKⅡmRNA相对表达量,Western blot法检测大鼠心房肌CAV1.2、CaM、CaMKⅡ及下游蛋白RyR2、P-RyR2蛋白表达情况。结果与空白组相比,模型组大鼠均出现典型房颤心电图(P<0.01),心房肌细胞肌丝排列絮乱、线粒体嵴断裂、呈“空泡样”改变,CAV1.2 mRNA及蛋白表达显著降低(P<0.01),CaM、CaMKⅡmRNA及蛋白表达显著升高(P<0.01),P-RyR2蛋白表达显著升高(P<0.01),RyR2蛋白表达无差异(P>0.05)。与模型组相比,中药复方组房颤持续时间降低(P<0.05);肌丝排列相对整齐,线粒体结构相对完整;CAV1.2 mRNA及蛋白表达升高(P<0.05),CaM、CaMKⅡmRNA及蛋白表达下降(P<0.01),下游蛋白P-RyR2表达降低(P<0.01),RyR2蛋白表达无差异(P>0.05)。结论祛痰化瘀中药复方可缩短大鼠房颤持续时间,抑制心房肌细胞超微结构损伤,其作用机制可能与调控CAV1.2/CaM/CaMKⅡ信号通路表达,改善钙调控紊乱有关。Objective To explore the mechanism of the treatment of atrial fibrillation(AF)with traditional Chinese medicine compound of eliminating phlegm and removing blood stasis.Methods Sixty male SD rats were randomly divided into the blank group and the model group.Ach(66μg·mL^(-1))-CaCl_(2)(10 mg·mL^(-1))was injected into the tail vein for 7 consecutive days to establish the rat AF model.The rats with successful modeling were randomly divided into model group,high,medium and low dose groups of Chinese herbal compound and verapamil group.The high,medium and low dose groups of Chinese herbal compound were given 12.38 mg,6.18 mg and 3.10 mg·kg^(-1)·d^(-1)Chinese herbal compound for removing phlegm and removing blood stasis solution by gavage,while the verapamil group was given 8.31 mg·kg^(-1)·d^(-1)verapamil solution by gavage,and the blank group and model group were given equal volume distilled water by gavage.During this period,the rats were still given tail vein injection for 14 consecutive days.The duration of atrial fibrillation in lead II of rats was measured by electrophysiological recorder,the ultrastructural changes of rat atrial muscle were observed by transmission electron microscope,the relative expression of CAV1.2,CaM,CaMKⅡmRNA in rat atrial muscle was detected by RTPCR,and the expression of CAV1.2,CaM,CaMKⅡand downstream proteins RyR2,P-RyR2 in rat atrial muscle was detected by Western blot.Results Compared with the blank group,the rats in the model group showed typical atrial fibrillation ECG.Compared with the model group,the duration of atrial fibrillation in the compound Chinese medicine group decreased.The arrangement of myofilaments was relatively neat,and the structure of mitochondria was relatively complete;CAV1.2 mRNA and protein expression increased(P<0.05),CaM,CaMKⅡmRNA and protein expression decreased(P<0.01),downstream protein P-RyR2 expression decreased(P<0.01),RyR2 protein expression had no difference(P>0.05).Conclusion The Chinese herbal compound for removing phlegm and removing

关 键 词：心房颤动 钙离子 CAV1.2/CaM/CaMKⅡ通路 祛痰化瘀中药复方 

分 类 号：R285.5[医药卫生—中药学]