基于TRPV1对动脉血管平滑肌的自噬作用探讨人参汤治疗AS的作用机制  

Mechanism of Renshentang in Treatment of Atherosclerosis Based on Autophagic Effect of TRPV1 on Arterial Smooth Muscle

作　　者：亓玉婕 何湛湛 杨桢[2] 陶旭光[1] 褚策 袁雨露 陈香云 丁薇 赵培彰 赵红霞[1] 汪文来[1] QI Yujie;HE Zhanzhan;YANG Zhen;TAO Xuguang;CHU Ce;YUAN Yulu;CHEN Xiangyun;DING Wei;ZHAO Peizhang;ZHAO Hongxia;WANG Wenlai(Institute of Basic Theory of Traditional Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China;Beijing University of Chinese Medicine,Beijing 102488,China;West China Clinical Medical College,Sichuan University,Chengdu 610041,China)

机构地区：[1]中国中医科学院中医基础理论研究所,北京100700 [2]北京中医药大学,北京100029 [3]四川大学华西临床医学院,成都610041

出　　处：《中国实验方剂学杂志》2023年第12期55-62,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：中国中医科学院科技创新工程黑地黄丸重大攻关项目(CI2021A00606);中国中医科学院自主选题项目(YZX202237,YZX202241,YZX202246,YZ2020042,YZ202225,YZ2020019)。

摘　　要：目的:基于香草酸受体亚型1(TRPV1)对动脉血管平滑肌的自噬作用探讨《金匮要略》人参汤治疗动脉粥样硬化(AS)的作用机制。方法:SPF级8周龄雄性C57BL/6J小鼠14只作为正常组,8周龄载脂蛋白E基因敲除(Apo E^(-/-))小鼠70只作为模型组。模型组采用高脂饲料喂食8周制备AS小鼠模型,随后随机分为模型组、人参汤低、中、高剂量(2.715、5.43、10.68 g·kg^(-1)·d^(-1))组和辛伐他汀(0.02 g·kg^(-1)·d^(-1))组,连续给药8周。8周后,取血清,采用检测试剂盒测定血清总胆固醇(CHO)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平,观察小鼠血脂水平的变化;取主动脉,采用苏木素-伊红(HE)染色观察主动脉根部整体的病理情况,采用油红O染色检测主动脉斑块内脂质沉积情况,并计算主动脉根部面积占管腔面积百分比;采用蛋白免疫印迹法(Western blot)检测小鼠主动脉组织中TRPV1、腺苷酸活化蛋白激酶(AMPK)、磷酸化腺苷酸活化蛋白激酶(p-AMPK)及自噬效应蛋白-1(Beclin-1)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)、微管相关蛋白1轻链Ⅰ(LC3Ⅰ)的表达情况。结果:与正常组比较,模型组小鼠血清中CHO、TG、LDL-C水平显著升高,HDL-C水平显著降低,主动脉根部斑块面积显著增多(P<0.01);与模型组比较,人参汤各剂量组可明显降低AS模型小鼠血清中CHO、TG、LDL-C水平(P<0.05,P<0.01),人参汤低、中剂量组最为显著(P<0.01)。与正常组比较,辛伐他汀组、人参汤低、中、高剂量组均能明显减少主动脉根部斑块面积(P<0.05,P<0.01),其中人参汤高剂量组效果减轻斑块效果最为显著(P<0.01)。与正常组比较,模型组小鼠的TRPV1、p-AMPK/AMPK、Beclin-1、LC3Ⅱ/LC3Ⅰ的相对表达量明显降低(P<0.05,P<0.01);与模型组比较,人参汤中、高剂量组TRPV1、p-AMPK/AMPK、Beclin-1、LC3Ⅱ/LC3Ⅰ的相对表达量均明显升高(P<0.05,P<0.01)。结论:《金匮要略》人参�Objective:To investigate the mechanism of Renshentang,recorded in Synopsis of Golden Chamber,in the treatment of atherosclerosis (AS) based on the autophagic effect of transient receptor potential vanilloid subtype 1 (TRPV1) on arterial smooth muscle.Method:Fourteen SPF-grade 8-week-old male C57BL/6J mice were assigned to the normal group and 70 8-week-old apolipoprotein E knockout (Apo E~(-/-))mice were assigned to the experimental group.The AS model was induced by a high-fat diet in the mice in the experimental group for eight weeks.The model mice were then randomly divided into model group,low-,medium-,and high-dose Renshentang groups (2.715,5.43,and 10.68 g·kg^(-1)·d^(-1)),and simvastatin group(0.02 g·kg^(-1)·d^(-1)).Drug treatment lasted eight weeks.Serum was taken and serum total cholesterol (CHO),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),and high-density lipoprotein cholesterol(HDL-C) levels were measured by assay kits to observe the changes in lipid levels in mice.The aorta was stained with hematoxylin-eosin (HE) to observe the overall pathology of the aortic root and oil red O staining was used to detect the lipid deposition in the aortic plaque and calculate the percentage of the aortic root area to the lumen area.The protein expression of TRPV1,adenylate-activated protein kinase (AMPK),phosphorylated AMPK(p-AMPK),autophagy effector-1 (Beclin-1),and microtubule-associated protein 1 light chain 3 (LC3Ⅱ)in mouse aortic tissues was determined by Western blot.Result:Compared with the normal group,the model group showed increased serum CHO,TG,and LDL-C levels,decreased HDL-C,and increased aortic root plaque area (P<0.01).Compared with the model group,the Renshentang groups showed decreased levels of CHO,TG,and LDL-C in serum (P<0.05,P<0.01),especially in the low-and medium-dose Renshentang groups (P<0.01).Compared with the normal group,the simvastatin group and the Renshentang groups showed reduced aortic root plaque area (P<0.05),especially in the high-dose Renshentang group (P<0.0

关 键 词：《金匮要略》人参汤 动脉粥样硬化 香草酸受体亚型1 自噬 载脂蛋白E基因敲除小鼠 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33