黄连解毒汤对ApoE-/-小鼠动脉粥样硬化炎症及主动脉斑块稳定性作用的机制研究

目的 观察黄连解毒汤对ApoE-/-小鼠主动脉粥样硬化炎症及斑块稳定性的影响，探讨黄连解毒汤治疗主动脉斑块稳定性的作用机制。方法 将36只ApoE-/-小鼠随机分为空白组，模型组，黄连解毒汤低、中、高剂量组，阿托伐他汀组，每组6只。模型组及给药组ApoE-/-小鼠给予高脂饮食构建主动脉粥样硬化模型8周后，黄连解毒汤低、中、高剂量组小鼠分别按10、20、40 g/（kg·d）剂量给予黄连解毒汤；阿托伐他汀组小鼠按10 mg/（kg·d）剂量给予阿托伐他汀；空白组、模型组给予等量的蒸馏水。每日灌胃1次，共8周。第16周末采集标本，采用苏木精—伊红染色观察各组小鼠主动脉内膜的组织形态；半自动生化分析仪检测各组小鼠血清总胆固醇（total cholesterol，TC）、三酰甘油（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；采用ELISA法检测各组白细胞介素-1β（interleukin-1β，IL-1β）、白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-10（interleukin-10，IL-10）含量；组织学特殊病理染色技术测定斑块内巨噬细胞含量、α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）细胞含量、斑块内胶原纤维含量及斑块内脂质含量，从而计算斑块不稳定指数（vulnerability index，VI）。结果 黄连解毒汤能明显改善小鼠主动脉管壁的病理变化。与空白组比较，模型组小鼠血清TC、TG、LDL-C水平均显著上升（P＜0.05），HDL-C水平显著下降（P＜0.05）；IL-1β、IL-6、TNF-α含量均显著增加（P＜0.05），IL-10含量显著减少（P＜0.05）；VI显著增加（P＜0.05）。与模型组比较，黄连解毒汤中、高剂量组小鼠血清TG、TC、LDL-C水平均显著下降（P＜0.05），HDL-C水平显著上升（P＜0.05）；IL-1β、IL-6、TNF-α含量均显著减少（P＜0.05），IL-10含量显著增加（P＜0.05）；各给药组小鼠主动脉内膜斑块VI均显著减少（P＜0.05）。黄连解毒汤高剂量组改善上述指标的效果与阿托伐他汀组相当（P＞0.05）。结论 黄连解毒汤对动脉粥样硬化ApoE-/-小鼠主动脉血管内炎症反应及斑块稳定性具有一定的改善作用，且呈剂量依赖性，其机制可能与降低血脂水平、缓解炎症损伤有关。

Mechanism of Action of Huanglian Jiedu Decoction on Atherosclerotic Inflammation and Aortic Plaque Stability in ApoE-/- Mice

Objective To investigate the effect of Huanglian Jiedu Decoction on aortic atherosclerotic inflammation and plaque stability in ApoE-/- mice,as well as the mechanism of action of Huanglian Jiedu Decoction in the treatment of aortic plaque stability.Methods A total of 36 ApoE-/- mice were randomly divided into blank group,model group,atorvastatin group,and low-,medium-,and high-dose Huanglian Jiedu Decoction groups,with 6 mice in each group.The ApoE-/- mice in the model group and the administration groups were given high-fat diet to establish a model of aortic atherosclerosis; after 8 weeks of modeling,the mice in the low-,medium-,and high-dose Huanglian Jiedu Decoction groups were given Huanglian Jiedu Decoction at doses of 10,20 and 40 g/(kg·day), respectively,those in the atorvastatin group were given atorvastatin at a dose of 10 mg/(kg·day),and those in the blank group and the model group were given an equal volume of distilled water,by gavage once a day for 8 weeks.After related samples were collected at the end of week 16,hematoxylin-eosin staining was used to observe the pathological and structural changes of aortic intima;a semi-automatic biochemical analyzer was used to measure the serum levels of total cholesterol (TC),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),and high-density lipoprotein cholesterol (HDL-C);ELISA was used to measure the content of interleukin-1β (IL-1β),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α),and interleukin-10 (IL-10);the special histopathological staining technique was used to measure the content of macrophages,alpha-smooth muscle actin (α-SMA)-positive cells,collagen fibers,and lipids in plaques,so as to calculate plaque vulnerability index (VI).Results Huanglian Jiedu Decoction significantly improved the pathological changes of the aortic wall.Compared with the blank group,the model group had significant increases in the serum levels of TC,TG,and LDL-C (P<0.05),a significant reduction in the serum level of HDL-C (P<0.05),significant increases in the content of IL-1β,IL-6,and TNF-α(P<0.05),a significant reduction in the content of IL-10 (P<0.05), and a significant increase in VI (P<0.05).Compared with the model group,the medium- and high-dose Huanglian Jiedu Decoction groups had significant reductions in the serum levels of TG,TC, and LDL-C (P<0.05),a significant increase in the serum level of HDL-C (P <0.05),significant reductions in the content of IL-1β,IL-6,and TNF-α (P<0.05),and a significant increase in the content of IL-10 (P<0.05), and all administration groups had a significant reduction in the VI of aortic intima plaques (P<0.05). The high-dose Huanglian Jiedu Decoction group and the atorvastatin group had similar improvements in the above indicators (P>0.05).Conclusion Huanglian Jiedu Decoction can improve inflammatory response and plaque stability in the aorta vessels of ApoE-/- mice with AS in a dose-dependent manner,possibly by reducing the levels of blood lipids and alleviating inflammatory injury.