摘要
目的:观察葱白提取物对心肌梗死模型大鼠心肌纤维化及转化生长因子β1蛋白(TGF-β1)、Smad2蛋白表达的影响,探讨其对心肌纤维化的抑制作用及可能机制。方法:结扎大鼠冠脉左前降支,建立急性心肌梗死(AMI)大鼠模型,造模成功后将其随机分为模型对照组,葱白提取物小、中、大剂量组(简称葱白小、中、大剂量组)及培哚普利组,另设假手术组,只穿线不结扎,每组9只大鼠。造模1周后,以0.9%氯化钠溶液给假手术组及模型对照组灌胃,其余4组以不同剂量相应药物灌胃;连续灌胃12周后,酶联免疫吸附测定法(ELISA)检测6组大鼠血清脑钠素(BNP)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)含量,取左心室心肌做苏木精-伊红(HE)染色、Masson染色,并采用Western blotting和实时荧光定量聚合酶链反应(PCR)分别检测心肌组织TGF-β1、Smad2蛋白和mRNA表达。结果:HE染色显示模型对照组大鼠心肌细胞水肿、坏死明显,结构破坏,排列凌乱;各治疗组大鼠心肌细胞水肿、坏死数量均减少,局部可见完整心肌细胞,炎症细胞浸润面积缩小。Masson染色显示模型对照组大鼠心肌组织破坏严重,心肌细胞残存较少,胶原纤维增加明显,药物治疗后,大鼠完整心肌细胞增多,胶原纤维不同程度减少,葱白大剂量组与培哚普利组抑制心肌纤维化的作用优于葱白小剂量和中剂量组。Image J图像分析Masson切片显示葱白大剂量组与培哚普利组心肌组织原容积分数(CVF)值下降明显。ELISA检测显示模型对照组BNP、TNF-α、IL-6含量较假手术组升高(P<0.01);与模型对照组相比,各治疗组BNP、TNF-α、IL-6含量均有所下降(P<0.01)。Western blotting和PCR检测结果显示,模型对照组较假手术组TGF-β1、Smad2蛋白表达升高明显(P<0.01);与模型对照组相比,葱白大剂量组、葱白中剂量组和培哚普利组TGF-β1、Smad2蛋白表达显著降低(P<0.01)。结论:葱白提取物可抑制心肌梗死后心力衰竭大鼠的心肌纤维化,其作用机制可能与减轻炎症反应、抑制TGF-β1/Smads信号传导通路有关。
Objective:To observe the effect of Allium Stalk extract on myocardial fibrosis and TGF-β1 and Smad2 protein expression in rats after myocardial infarction,and to explore its inhibitory effect on myocardial fibrosis and its possible mechanism.Methods:Acute myocardial injury(AMI)model was established by ligating the left anterior descending coronary artery in rats.The model rats were randomly divided into blank model group,perindopril group,low-dose group,medium-dose group and high-dose group.In addition,a sham operation group was set up without ligation.After 1 week,normal saline was given to the sham group and the blank model group,and the other four groups were given different doses of corresponding drugs.After 12 weeks of continuous intragastric administration,serum levels of BNP、TNF-αand IL-6 in the six groups of rats were detected.Left ventricular myocardium was stained with HE and Masson staining,and TGF-β1,Smad2 protein and mRNA expression in myocardium were detected by Western blotting and real-time fluorescence quantitative PCR.Results:The HE staining showed obvious edema and necrosis of myocardial cells in the blank model group,and the structure was damaged and arranged in disorder.The number of edema and necrosis of myocardial cells in all treatment groups was reduced,complete myocardial cells could be seen locally,and the infiltration area of inflammatory cells was reduced.Masson staining showed that the myocardial tissue in the blank model group was severely damaged,the myocardial cell survival was small,and the collagen fiber increased significantly.After drug treatment,the rat myocardial cell integrity increased,and the collagen fiber decreased to different degrees.The effect of the high-dose group and perindopril group on inhibiting myocardial fibrosis was better than that of the low-dose and medium-dose group.Analysis of Image J Masson section showed that CVF value decreased significantly in the high-dose group and perindopril group.The ELISA assay showed that the levels of BNP,TNF-αand IL-6 in the blank model group were higher than those in the sham group(P<0.01).Compared with the blank model group,the content of BNP,TNF-αand IL-6 decreased in each treatment group(P<0.01).Western blot and PCR results showed that TGF-β1 and Smad2 protein expressions were significantly increased in the blank model group compared with the sham operation group(P<0.01).Compared with blank model group,high dose group,medium dose group and without perindopril captopril group TGF-β1 and Smad2 protein expression significantly reduced(P<0.01).Conclusion:The inhibitory effect of Allium Stalk extract on myocardial fibrosis in rats with heart failure after myocardial infarction may be related to the reduction of inflammatory response and the inhibition of TGF-β1/Smads signaling pathway.
作者
赵辉
邢若丹
谢婷婷
马威
杨文强
余志华
田立群
吴勇宏
柯于鹤
ZHAO Hui;XING Ruodan;XIE Tingting;MA Wei;YANG Wenqiang;YU Zhihua;TIAN Liqun;WU Yonghong;KE Yuhe(Department of Geriatric,Wuhan Hospital of Traditional Chinese and Western Medicine,Wuhan 430022,China;First Clinical College,Hubei University of Chinese Medicine,Wuhan 430065,China;Department of Cardiology,Wuhan Hospital of Traditional Chinese and Western Medicine,Wuhan 430022,China)
出处
《医药导报》
CAS
北大核心
2020年第5期604-609,共6页
Herald of Medicine
基金
湖北省自然科学基金资助项目(2017CFB783,2018CFC801)
湖北省卫健委中医药管理局课题(ZY2019M045)。
