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 choles-terol,TC)、三酿甘油(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(in-terleukin-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(vul-nerability 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、1L-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 sta-bility.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 aor-tic 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 lev-els 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 sig-nificant 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 inflam-matory response and plaque stability in the aorta vessels of ApoE-/- mice with AS in a dose-dependent manner,possibly by re-ducing the levels of blood lipids and alleviating inflammatory injury.
Huanglian Jiedu DecoctionAtherosclerosisBlood lipidInflammatory cytokinesPlaque vulnerability index
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