摘要
目的通过观察芪仙颗粒对缺血性心肌病模型大鼠心肌鸟核苷耦联激活性蛋白α亚基(Gsα)和抑制性蛋白α亚基(Giα)含量及磷脂酰肌醇4,5-二磷酸(PIP2)-1,4,5-三磷酸肌醇(IP3)-Ca^(2+)-钙调神经磷酸酶(CaN)信号通路的影响,探讨其抗缺血性心肌重构的机制。方法以异丙肾上腺素(ISO)腹腔注射构建缺血性心肌病模型,随机分为对照组、ISO模型组、芪仙颗粒低剂量组和芪仙颗粒高剂量组。酶联免疫吸附法测定心肌组织匀浆CaN、PIP2、IP3含量,超声心动图观察心肌结构,苏木素-伊红染色观察心肌组织病理学结构变化,RT-PCR法检测心肌组织Gsα、Giα、IP3、1,4,5-三磷酸肌醇受体(IP3R)、CaN mRNA表达,WesternBlot检测左室心肌组织Gsα、Giα蛋白的表达。结果芪仙颗粒可抑制ISO诱导的大鼠心肌组织匀浆PIP2、IP3、CaN升高(P<0. 05);病理结果显示,芪仙颗粒可减轻ISO诱导的心肌坏死、纤维化和炎性细胞浸润的程度和范围;超声心动图结果显示,芪仙颗粒大剂量组左心室舒张末期直径(LVEDD)和左心室收缩末期直径(LVESD)明显降低(P <0.05);RT-PCR结果显示,芪仙颗粒较ISO组大鼠心肌组织Gsα、IP3、IP3R、CaNmRNA水平均有显著降低(P<0. 01),Giα表达水平升高(P<0. 05);Western Blot结果显示,芪仙颗粒可明显增加Giα蛋白的表达、减少Gsα蛋白的表达(P <0.05)。结论 ISO诱导的心肌重构机制与心肌G蛋白偶联相关的Ca^(2+)信号通路传导紊乱有关,抑制G蛋白-PIP2-IP3-Ca^(2+)信号途径可能是芪仙颗粒抗缺血性心肌重构的分子生物学机制之一。
Objective To investigate the inhibitory effect of Qixian granules on isoproterenol (ISO)-induced myocardial remodeling rats and explore its effect on Gsα/Giα-PIP2-IP3 Ca^2+ signaling pathway. Methods The ischemic cardiomyopathy model was established by intraperitoneal injection of ISO,and then the rats were randomly divided into control group,ISO model group,high dosage Qixian granules group and low dosage Qixian granules group. After intervention for 4 weeks,the pathological changes of myocardium were observed by HE staining;the structure of myocardium was observed by echocardiography;levels of CaN,PIP2 and IP3 were detected by ELISA;the mRNA expression levels of Gsα,Giα,IP3,IP3R and CaN in left ventricular tissues were detected by RTPCR;and the protein expression levels of Gsα and Giα in left ventricular tissues were detected by Western Blot. Results Qixian granules could significantly inhibit the increase of PIP2,IP3 and CaN levels in myocardial tissue homogenate of rats with ISO-induced myocardial remodeling (P<0.05), alleviate ISO-induced myocardial necrosis,fibrosis and inflammatory cell infiltration,reduce ISO-induced increase of LVEDD and LVESD (P<0.05). RT-PCR results showed that Gsα, IP3, IP3R, and CaN expression levels were increased and Giα expression levels were decreased in the myocardial tissues of ISO group rats; and after Qixian granules administration,the above expression levels were decreased (Gsα,IP3,IP3R,and CaN,P<0.01) or incressed (Giα,P<0.05). Western Blot results showed that Qixian granules could significantly increase the expression of Giα protein and decrease the expression of Gsα protein (P<0.05). Conclusion Our findings indicated that ISO induced myocardial remodeling mechanism is related to myocardial G protein coupling Ca^2+ signal pathway disorders; inhibiting the G protein- PIP2-IP3 Ca^2 + signaling pathways may be one of the molecular mechanisms of Qixian granules against ischemic myocardial remodeling.
作者
王新东
祁晓霞
WANG Xindong;QI Xiaoxia3(Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine,Nanjing 210028 Jiangsu,China;Department of Cardiology,Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing 210028 Jiangsu, China;Office of Science and Technology Administration,Nanjing University of Chinese Medicine,Nanjing 210026 Jiangsu,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2018年第6期719-724,共6页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
江苏省自然科学基金青年基金资助项目(BK20161078)
南京市科技项目(201715068)
国家自然科学基金青年基金资助项目(NO.81403386)
