益气活血法对舒张性心力衰竭大鼠心肌细胞钙稳态的影响  

Effects of qi-nourishing and blood-activating method on myocardium calcium homeostasis in rats with diastolic heart failure

作　　者：刘静[1] 徐江林 石晓路[3] 刘璐 李思耐[5] 崔海峰[3] 黄雁超 武乾[3] 万洁[1] 周鹍[1] 崔晓云[1] 李岩[1] 逯金金[1] 李冬 高群 贾文浩 孙明杰[3] 林谦 LIU Jing;XU Jianglin;SHI Xiaolu;LIU Lu;LI Sinai;CUI Haifeng;HUANG Yanchao;WU Qian;WAN Jie;ZHOU Kun;CUI Xiaoyun;LI Yan;LU Jinjin;LI Dong;GAO Qun;JIA Wenhao;SUN Mingjie;LIN Qian(Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100078,China;Dongzhimen Hospital,Beijing University ofChinese Medlicine,Bejing 1007,Chinma;Beijing Key Laboratory of TCM Basic Research on Prevention and Treatment of MajorDisease,Experimental Research Center,China Academy of Chinese Medical Science,Beijing 100700,China;Shunmyi Hospital,Beijing Traditional Chinese Medicime Hospital,Beijing 101300,China;Beijing Hospital of Traditional Chinese Medicine,CapitalMedlicalUniversity,Beijing Institute of Traditional Chinese Medicine,Bejing 100010,China;Bejing Univerity of Chinese Melicine ThirdAffiliated Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学东直门医院 [3]中国中医科学院实验研究中心北京市重点实验室中医药防治重大疾病基础研究实验室 [4]北京市中医院顺义医院 [5]首都医科大学北京中医医院 [6]北京中医药大学第三附属医院

出　　处：《北京中医药大学学报》2022年第2期165-175,共11页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(No.81373779);国家自然科学基金青年科学基金项目(No.81903993)。

摘　　要：目的研究不同比例益气活血药对舒张性心力衰竭(DHF)大鼠的疗效,探讨其对心肌细胞钙稳态的影响机制。方法采用改良缩窄腹主动脉术制备压力负荷致DHF大鼠模型。将大鼠分为假手术组、模型组、益气组(黄芪3 g、党参1.5 g)、益气活血1∶1组(黄芪3 g、党参1.5 g、丹参3 g、三七1 g)、益气活血2∶1组(黄芪6 g、党参3 g、丹参3 g、三七1 g),每组20只,连续干预12周。治疗4、12周进行心功能检测,测量舒张期左室前壁厚度、舒张期左室后壁厚度、舒张期左室内径、左室射血分数和左室短轴缩短率及舒张早期跨二尖瓣血流速度(E)与舒张早期二尖瓣环运动幅度(e)比值(E/e);12周进行血流动力学检测,分别在静息状态和盐酸多巴胺负荷状态下记录左室压力下降速率、左室舒张末期压力(LVEDP)以评价左室舒张功能,记录左室压力上升速率,左室收缩压力(LVESP)以评价左室收缩功能,并记录心率及颈动脉压力;12周后处死大鼠,检测心肌细胞舒缩功能及钙转运情况,测量心肌细胞的收缩速率、舒张速率、收缩50%时间(CT50)和舒张50%时间(RT50);测定钙释放速率、钙回摄速率、钙释放50%时间(CaT50-on)和钙回摄50%时间(CaT50-off),分析钙瞬态数据;Western blot法检测钙转运相关蛋白钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)、蛋白激酶A(PKA)、16-丝氨酸位点磷酸化受磷蛋白(PLBS16)、17-苏氨酸位点磷酸化受磷蛋白(PLBT17)表达水平。结果干预12周后,全部治疗组均改善大鼠舒缩功能LVEDP、RT50、CT50,益气活血2∶1组改善舒张早期二尖瓣血流速度E峰/舒张早期二尖瓣环纵向运动速率e峰(E/e)比值、多巴胺负荷下LVEDP和逆转心室重构(降低左室前壁和后壁厚度)。在细胞水平,全部治疗组均缩短心肌细胞CT50、RT50、CaT50-on和降低CaMKⅡ表达,两组益气活血治疗组还可降低PKA过表达,益气活血2∶1组可降低CaT50-off和改善PLBS16低磷酸化Objective To explore the efficacy of nourishing qi(NouQ) and activating blood(ActB) herbal combination at different ratios on diastolic heart failure(DHF) rats by targeting calcium homeastasis. Methods DHF rats were induced by abdominal aortic constriction and grouped into NouQ(AM+CP), NouQ vs ActB=1∶1(AM+CP vs SM+PP=1∶1), or NouQ vs ActB=2∶1(AM+CP vs SM+PP=2∶1). Heart function, hemodynamics, cardiomyocytic shortening and calcium transient were measured after 4/12 weeks of treatment. CaMKⅡ, protein kinase A(PKA) and their responding phosphorylation proteins PLB T17 and PLB S16 were examined by using Western blot assay.Results Although all NouQ + ActB groups showed improvements on diastolic function(lower end-diastolic left ventricular pressure(LVEDP) at resting status;shorter time to 50% relaxation of cardiomyocytes) and systolic function(shorter time to 50% contraction of cardiomyocytes), only NouQ vs ActB=2∶1 group decreased E/e ratio, LVEDP under exercise stress and reversed LV remodeling(lower LV anterior wall thickness and LV posterior wall thickness). At cellular level, all treatment groups showed shorter time to 50% contraction, shorter time to 50% relaxation, shorter time to 50% Ca;release and reduction of CaMKⅡ expression;all NouQ + ActB treatment groups illustrated additional effects on inhibiting PKA overexpression, but only 2∶1 group illustrated shorter time to 50% Ca;uptake and reversed hypo-phosphorylation of PLB S16. Conclusion The combination of NouQ herbs(AM+CP) and ActB herbs(SM+PP) at a ratio of 2∶1 have a better therapeutic effect than that of a ratio of 1∶1 and NouQ herbs on DHF, and this effect may work through regulating Ca;handling.

关 键 词：舒张性心力衰竭 益气活血法 钙稳态 大鼠 

分 类 号：R285.5[医药卫生—中药学]