黄芪-当归配伍对小鼠动脉粥样硬化模型血管损伤的影响  

Effect of Astragalus-Angelica combination on vascular injury in a mouse model of atherosclerosis

作　　者：李菀榆 龙清吟 傅馨莹 马露 谭维 李艳玲 徐顺洲 张伟 邓常清 LI Wanyu;LONG Qingyin;FU Xinying;MA Lu;TAN Wei;LI Yanling;XU Shun-zhou;ZHANG Wei;DENG Changqing(School of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Hunan Key Laboratory of Inte-grated Chinese and Western Medicine for Prevention and Treatment of Heart and Brain Diseases,Changsha 410208,China)

机构地区：[1]湖南中医药大学中西医结合学院,中西医结合心脑疾病防治湖南省重点实验室,湖南长沙410208

出　　处：《中国病理生理杂志》2023年第9期1586-1595,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81874406,No.82174218);湖南省自然科学杰出青年基金项目(No.2020JJ2024)。

摘　　要：目的:从血管壁细胞凋亡和焦亡方面来探讨黄芪-当归配伍对小鼠动脉粥样硬化(atherosclerosis,AS)模型血管壁细胞损伤的影响。方法:将雄性载脂蛋白E基因敲除(apolipoprotein E gene knockout,ApoE-/-)小鼠随机分为模型组、黄芪-当归配伍低剂量组、黄芪-当归配伍中剂量组、黄芪-当归配伍高剂量组和阿托伐他汀组,以同周龄雄性C57BL/6J小鼠为对照组,每组5只。除对照组外均给予高脂饲料喂养12周构建AS模型,模型成功后灌胃给予药物。HE染色观察主动脉内膜增生厚度;全自动生化分析仪检测总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-c)及高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-c)水平;ELISA法检测血浆白细胞介素1β(interleukin-1β,IL-1β)和IL-18含量;免疫荧光法、RT-qPCR法及Western blot法检测血管壁细胞凋亡和焦亡相关因子B细胞淋巴瘤蛋白2(B-cell lympho-ma-2,Bcl-2)、Bcl-2关联X蛋白(Bcl-2-associated X protein,Bax)、活化的胱天蛋白酶3(cleaved caspase-3)、核苷酸结合寡聚结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)、cleaved cas-pase-1和gasdermin D-N(GSDMD-N)的mRNA及蛋白表达,评价药物对AS病变血管壁细胞损伤的作用。结果:高脂喂养可成功建立AS模型。与对照组比较,模型组血浆TC、TG和LDL-c含量显著升高(P<0.01),HDL-c含量显著降低(P<0.01),血管内膜增厚(P<0.01),血浆IL-1β和IL-18含量显著增加(P<0.01),血管壁细胞Bax、cleaved cas-pase-3、NLRP3、cleaved caspase-1和GSDMD-N表达增加(P<0.01),Bcl-2表达降低(P<0.01)。与模型组比较,黄芪-当归配伍可降低血浆TC、TG和LDL-c含量(P<0.05),升高血浆HDL-c含量(P<0.05),减轻血管内膜增生程度(P<0.01),降低血浆IL-1β和IL-18含量(P<0.05),抑制血管壁细胞Bax、cleaved caspase-3、NLRP3、cleaved caspase-1和GSDMD-N表达(P<0.AIM:To investigate the effect of Astragalus-Angelica combination on cell injury in the vascular wall of mouse atherosclerosis(AS)model from perspectives of cell apoptosis and pyroptosis.METHODS:Male apolipo-protein E gene knockout(ApoE-/-)mice were randomly divided into model group,low-dose Astragalus-Angelica group,me-dium-dose Astragalus-Angelica group,high-dose Astragalus-Angelica group and atorvastatin group,with 5 mice in each group.The same number of male C57BL/6J mice at the same week of age were used as control.Except for the control group,all other animals were fed with high-fat feed for 12 weeks to construct AS model,and drugs were given by gavage af-ter successful modeling.Then,the thickness of aortic intimal hyperplasia was observed by HE staining.The total choles-terol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-c)and high-density lipoprotein cholesterol(HDL-c)levels were measured by a fully-automated biochemical analyzer,and the plasma interleukin-1β(IL-1β)and IL-18 levels were measured by ELISA.The mRNA and protein expression levels of cell apoptosis-and pyroptosis-related fac-tors,including B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cleaved caspase-3,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),cleaved caspase-1 and gasdermin D-N(GSDMD-N),were de-tected by immunofluorescence,RT-qPCR and Western blot assays.The combined results were used to evaluate the effect of target drugs on vascular wall cell injury in AS lesions.RESULTS:High-fat feeding induced AS model successfully.Compared with control group,the mice in model group showed increased plasma TC,TG and LDL-c levels(P<0.01),de-creased HDL-c level(P<0.01),thickened intima(P<0.01),increased plasma IL-1βand IL-18 levels(P<0.01),and elevated expression levels of Bax,cleaved caspase-3,NLRP3,cleaved caspase-1 and GSDMD-N,and reduced expression of Bcl-2 in the vascular wall(P<0.01).Compared with model group,treatment with Astragalus-Angelica combination down-regulated plasma TC,TG and LDL-c

关 键 词：动脉粥样硬化 黄芪-当归配伍 APOE-/-小鼠 细胞凋亡 细胞焦亡 

分 类 号：R363.2[医药卫生—病理学] R282.71[医药卫生—基础医学]