瓜蒌薤白半夏汤对心肌缺血模型大鼠心肌微血管生成与HIF-1α/VEGF相关通路的影响  

Effect of Gualou Xiebai Banxiatang on Myocardial Microangiogenesis and HIF-1α/VEGF-related Pathways in Myocardial Ischemia Model Rats

作　　者：陈文豪 孟维闪 李红 田维维 张琪[1] 谭颖颖[1] CHEN Wenhao;MENG Weishan;LI Hong;TIAN Weiwei;ZHANG Qi;TAN Yingying(Basic Medical College,Shaanxi University of Chinese Medicine,Xianyang 712046,China;The Second Affiliated Hospital of Henan University of Science and Technology,Luoyang 471000,China)

机构地区：[1]陕西中医药大学基础医学院,陕西咸阳712046 [2]河南科技大学第二附属医院,河南洛阳471000

出　　处：《中国实验方剂学杂志》2024年第17期1-9,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81873296,82074380)。

摘　　要：目的:探讨瓜蒌薤白半夏汤对缺血性心肌损伤大鼠心功能和心肌组织病理变化的影响,观察心肌微血管密度(MVD)、磷脂酰肌醇3-激酶(PI3K)雷帕霉素靶蛋白(mTOR)、缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)信号通路对心肌微血管生成的影响。方法:将70只雄性SD大鼠,随机选取6只为正常组,其余大鼠高脂饲料喂养,并注射盐酸异丙肾上腺素(ISO,80 mg·kg^(-1)·d^(-1),2 d),建立以高脂血症为基础的缺血性心脏病模型。模型成功后,随机分为模型组、瓜蒌薤白半夏汤高、中、低剂量组和美托洛尔组。其中,瓜蒌薤白半夏汤高、中、低剂量组分别给予瓜蒌薤白半夏汤水煎液10.42、5.21、2.61 g·kg^(-1)·d^(-1),美托洛尔组给予美托洛尔2.6 mg·kg^(-1)·d^(-1),正常组和模型组均给予等体积的生理盐水,持续28 d。干预治疗结束后取材,进行相关检测,收集血清用于检测心功能相关指标;取心室组织进行苏木素-伊红(HE)、马松(Masson)染色,观察其光镜下的病理变化;免疫组织化学法(IHC)检测血小板内皮细胞黏附分子(CD31)阳性表达数量;酶联免疫吸附测定法(ELISA)检测N末端B型脑钠肽前体(NT-proBNP)、VEGF的表达;蛋白免疫印迹法(Western blot)检测PI3K、mTOR、HIF-1α、VEGF蛋白表达水平。结果:与正常组比较,模型组大鼠血清乳酸脱氢酶(LDH)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、NT-proBNP及VEGF含量水平均显著升高(P<0.01);胶原容积分数(CVF)显著升高(P<0.01);MVD显著降低(P<0.01);PI3K、mTOR、HIF-1α、VEGF的蛋白表达水平均升高(P<0.05,P<0.01)。与模型组比较,美托洛尔组血清LDH、CK、CK-MB、NTproBNP显著降低(P<0.01),VEGF显著升高(P<0.01);CVF显著降低(P<0.01);MVD显著升高(P<0.01);PI3K、mTOR、VEGF蛋白表达水平均显著升高(P<0.01),其中HIF-1α蛋白表达水平变化差异无统计学意义。与模型组比较,瓜蒌薤白半夏汤高、中剂量组血清LDH、CK、CK-MB及NTObjective:To investigate the effect of Gualou Xiebai Banxiatang on cardiac function and myocardial histopathological changes in rats with ischemic myocardial injury,and to observe the effect of myocardial microvascular density(MVD),phosphatidylinositol 3-kinase(PI3K),mammalian target of rapamycin(mTOR),hypoxia-inducible factor-1 alpha(HIF-1α),and vascular endothelial growth factor(VEGF)signaling pathways on myocardial microangiogenesis.Method:Seventy male SD rats were randomly selected,with six rats in the normal group.The remaining rats were fed a high-fat diet and injected with isoproterenol hydrochloride(ISO,80 mg·kg^(-1)·d^(-1),2 d)to induce a hyperlipidemia-based ischemic heart disease model.After successful modeling,the rats were randomly divided into the model group,high,medium,and low dose groups of Gualou Xiebai Banxiatang,and the metoprolol group.The high,medium,and low dose groups of Gualou Xiebai Banxiatang were given Gualou Xiebai Banxiatang at 10.42,5.21,2.61 g·kg^(-1)·d^(-1),respectively,while the metoprolol group was given metoprolol at 2.6 mg·kg^(-1)·d^(-1).Both the normal and model groups were given an equivalent volume of physiological saline for 28 days.After the intervention,relevant tests were conducted,and serum was collected to measure heart function-related indicators.Hematoxylin-eosin(HE)and Masson staining were performed on ventricular tissue to observe pathological changes under a light microscope.Immunohistochemistry(IHC)was used to detect the positive expression of platelet endothelial cell adhesion molecule(CD31).Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression of N-terminal pro-brain natriuretic peptide(NT-proBNP)and VEGF.Western blot was used to detect the protein expression levels of PI3K/mTOR/HIF-1α/VEGF.Result:Compared with the normal group,the model group showed significantly increased serum levels of LDH,CK,CK-MB,NT-proBNP,and VEGF(P<0.01),significantly increased collagen volume fraction(CVF)(P<0.01),significantly decreased MVD(P<0.01),and ele

关 键 词：瓜蒌薤白半夏汤 缺血性心脏病 内皮细胞功能 心肌纤维化 血管生成 缺氧诱导因子-1α(HIF-1α) 血管内皮生长因子(VEGF) 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5R541