黄连解毒汤通过抑制ApoE-/-小鼠巨噬细胞极化和炎症减轻高脂饮食诱导的动脉粥样硬化

Huanglian Jiedu Decoction reduce high-fat diet-induced atherosclerosis by inhibiting macrophage polarization and inflammation in ApoE-/- mice

目的 探讨黄连解毒汤(HLJDD)抑制巨噬细胞极化和炎症对ApoE-/-小鼠动脉粥样硬化(AS)的影响及作用机制。方法 构建ApoE-/-小鼠AS模型,随机分为模型组、黄连解毒汤高剂量(HLJDD-H)组和黄连解毒汤低剂量(HLJDD-L)组,另设C57BL/6J小鼠为正常(Control)组,每组6只,分别给予相应浓度药物及生理盐水灌胃,每日1次,连续6 w。给药结束后,收集小鼠血清和主动脉组织,全自动生化分析仪检测血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平;苏木素-伊红(HE)和油红O染色观察主动脉组织病理变化;酶联免疫吸附试验(ELISA)测定小鼠血清超敏C反应蛋白(hs-CRP)、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、IL-4和IL-10水平;荧光染色和免疫组化检测主动脉M1、M2型巨噬细胞标记物诱导型一氧化氮合酶(iNOS)、CD197和CD206分子的表达。结果 与Model组比较,HLJDD不同剂量组主动脉斑块面积和脂滴明显减少,血清TC、TG、LDL-C、hs-CRP、TNF-α和IL-6水平均显著降低(P<0.05,P<0.01),HDL-C、IL-4、IL-10水平显著升高(P<0.05,P<0.01);M1型巨噬细胞标记物iNOS、CD197表达显著降低(P<0.05),M2型巨噬细胞标记物CD206表达显著升高(P<0.05)。结论 HLJDD可通过调控巨噬细胞极化方向降低炎症反应,从而发挥抗AS作用。

Objective To investigate the effect and mechanism of Huanglian Jiedu Decoction(HLJDD) on ApoE-/-mice with atherosclerosis(AS) by inhibiting macrophage polarization and inflammation. Methods ApoE-/-mouse AS model was established, which was randomly divided into model group, Huanglian Jiedu Decoction high-dose(HLJDD-H) group, Huanglian Jiedu Decoction low-dose(HLJDD-L) group, and C57BL/6J mice were set as normal(Control) group, with 6 mice in each group.Each group was gaven corresponding concentrations of drugs and saline by gavage, once a day, consecutive for 6 weeks. After the administration, the mouse serum and aortic tissue were collected, and automatic biochemical analyzer was used to detect serum total cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterin(LDL-C) and high density lipoprotein cholesterol(HDL-C) levels;htoxylin-eosin(HE) and Oil Red O staining were used to observe the pathological changes of aortic tissue;enzyme linked immunosorbent assay(ELISA) was used to detect mouse serum hypersensitive C-reactive protein(hs-CRP), tumor necrosis factor(TNF)-α, interleukin(IL)-6, IL-4 and IL-10 levels;fluorescence staining and immunohistochemistry were used to detect the expression of aortic M1 and M2 macrophage markers inducible nitric oxide synthase(iNOS), CD197 and CD206 molecule.Results Compared with the model group, the aortic plaque area and lipid droplets in the HLJDD different dose group were significantly reduced, and the serum levels of TC, TG, LDL-C, hs-CRP, TNF-α and IL-6 were significantly reduced(P<0.05, P<0.01), the levels of HDL-C, IL-4 and IL-10 were significantly increased(P<0.05,P<0.01). The expressions of M1 macrophage markers iNOS and CD197 were significantly decreased(P<0.05), and the expression of M2 macrophage markers CD206 were significantly increased(P<0.05). Conclusions HLJDD could reduce the inflammatory response by regulating the polarization of macrophages, thereby exerting an anti-AS effect.