痰瘀同治方调控自噬对心肌缺血区和动脉粥样硬化斑块内血管新生不同作用的实验研究  

Experimental Study on the Different Effects of Tanyu Tongzhi Recipe Regulating Autophagy on Neovascularization in Myocardial Ischemic Area and Atherosclerotic Plaque

作　　者：蔡宏文[1] 李昕遥 周鑫斌[1] 李家英 陈晨 毛威[1] CAI Hongwen;LI Xinyao;ZHOU Xinbin(The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006,China)

机构地区：[1]浙江中医药大学附属第一医院,杭州310006

出　　处：《浙江中医药大学学报》2020年第7期652-656,共5页Journal of Zhejiang Chinese Medical University

基　　金：浙江省公益技术应用研究(实验动物)项目(2017C37182);浙江省中医药科研基金项目(2016ZA073、2019ZB037)。

摘　　要：[目的]明确痰瘀同治方对心肌缺血区和动脉粥样硬化(atherosclerosis,AS)斑块内血管新生的双向调节作用,探讨自噬在其中的调控机制。[方法]选取载脂蛋白E基因敲除(Apolipoprotein E gene knockout,ApoE-/-)小鼠20只,随机分为模型对照组和痰瘀同治(Tanyu Tongzhi,TYTZ)组,每组10只,饲喂高脂饲料;另取C57BL/6J小鼠10只为正常对照组,饲喂基础饲料。饲喂4周后,模型对照组和TYTZ组皮下注射异丙肾上腺素针100mg/kg,连续2d,正常对照组皮下注射等量0.9%氯化钠注射液。造模成功后4周,TYTZ组给予TYTZ方流浸膏2145mg/(kg·d),以蒸馏水稀释成0.4mL灌胃,模型对照组和正常对照组给予等量蒸馏水灌胃。给药6周后,用CD31免疫组化染色观察心肌缺血区和主动脉的新生血管密度。Western blot检测心肌组织和主动脉微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)Ⅱ/Ⅰ蛋白表达。[结果]免疫组化染色结果显示,模型对照组心肌组织和主动脉组织CD31表达均显著高于正常对照组(P<0.05);TYTZ组心肌组织CD31表达明显高于模型对照组(P<0.05),而主动脉组织CD31表达则明显低于模型对照组(P<0.05)。Western blot结果显示,模型对照组心肌组织和主动脉组织LC3Ⅱ/Ⅰ蛋白表达水平均明显高于正常对照组(P<0.05);TYTZ组心肌组织LC3Ⅱ/Ⅰ蛋白表达水平显著高于模型对照组(P<0.05),而主动脉组织LC3Ⅱ/Ⅰ蛋白表达则明显低于模型对照组(P<0.05)。[结论]TYTZ方在抑制AS斑块内血管新生的同时能够促进心肌缺血区的血管新生,该方对自噬水平的双向调节作用可能是其中的调控机制。[Objective]To clarify the two-way regulatory effect of Tanyu Tongzhi(TYTZ)Recipe on neovascularization in atherosclerotic plaque and myocardial ischemic area,and explore the regulatory mechanism of autophagy in this process.[Methods]Twenty Apolipoprotein E gene Knockout(ApoE-/-)mice were randomly divided into model control groups and TYTZ group,10 mice in each group,fed with high-fat diet.Ten C57 BL/6 J mice fed with basic diet were used as normal control group.After 4 weeks of feeding,model control group and TYTZ group were subcutaneously injected with isoproterenol at 100 mg/kg once a day for 2 days,while normal control group was subcutaneously injected with the same amount of 0.9%sodium chloride injection.Another 4 weeks later,TYTZ group was given 2145 mg/(kg·d)of TYTZ Recipe liquid extract(diluted to 0.4 mL with distilled water)by gavage,while the other two groups were given the equal amount of distilled water.After 6 weeks of administration,the density of neovascularization in myocardial ischemic area and aorta was detected by CD31 immunohistochemical staining,the protein expression of microlubule-assciated protein 1 light chain 3(LC3)Ⅱ/Ⅰin myocardial ischemic area and aorta were analyzed by Western blot.[Results]Immunohistochemical staining showed that the expression of CD31 in myocardium and aortic tissue of model control group was significantly higher than that of normal control group(P<0.05),the expression of CD31 in myocardium of TYTZ group was significantly higher than that in model control group(P<0.05),while the expression of CD31 in aortic tissue of TYTZ group was significantly lower than that in model control group(P<0.05).Western blot results showed that the level of LC3Ⅱ/Ⅰprotein expression in myocardium and aortic tissue of model control group was significantly higher than that of normal control group(P<0.05),and the level of LC3Ⅱ/Ⅰprotein expression in myocardium of TYTZ group was significantly higher than that in model control group(P<0.05),while the LC3Ⅱ/Ⅰprotein expression i

关 键 词：痰瘀同治方 自噬 血管新生 心肌缺血 动脉粥样硬化 CD31 LC3Ⅱ/Ⅰ 调控机制 

分 类 号：R331[医药卫生—人体生理学]