摘要
目的:观察活血方与阳与汤对高脂饮食导致动脉粥样硬化并发非酒精性脂肪肝的apoE^(-/-)小鼠全身、主动脉血管和肝脏免疫反应的影响。方法:10周龄雌性apoE^(-/-)小鼠给予高脂饮食,治疗组同时给予活血方或阳和汤干预18周,检测血脂肝功能水平、血浆细胞因子、外周血单核细胞比例、单核细胞亚型比例及其表面受体TLR4的表达、主动脉和肝脏的病理变化及炎性细胞因子表达。结果:18周高脂饮食导致apoE^(-/-)小鼠外周血TC和LDL水平显著增高,肝功能ALT和AST水平没有明显改变,外周血单核细胞比例、单核细胞炎症亚型Ly6C++及其表面受体TLR4的表达显著升高,血浆细胞因子IL-10、IL-1b、MCP-1、M-CSF和TNF-a水平升高;模型组小鼠主动脉血管检测到明显的AS斑块,肝脏油红O染色阳性;主动脉组织炎症因子TNF-a、MCP-1、IFN-γ、NOS2和IL-1b表达显著升高,肝组织炎症因子TNF-a、IL-6、MCP-1、IFN-γ、NOS2和IL-1b表达显著升高。18周的中药活血方和阳和汤干预并未引起血脂明显改善,肝功能ALT和AST水平也没有明显变化;但活血方降低外周血单核细胞及其Ly6C++亚型比例,活血方和阳和汤降低核细胞及其Ly6C++表面受体TLR4表达;对于血浆炎症因子,活血方降低IL-10、IL-1b、MCP-1和TNF-a水平,显著减低M-CSF水平,阳和汤仅对TNF-a水平略有降低;活血方和阳和汤干预18周肝脏油红O染色较模型组弱,活血方较阳和汤减弱更为显著。主动脉血管和肝脏实时定量PCR检测发现,活血方和阳和汤明显减低炎症因子TNF-a、MCP-1、IFN-γ、NOS2和IL-1b表达,活血方降低效果更为显著。结论:长期高脂饮食导致apoE^(-/-)小鼠动脉粥样硬化并发非酒精性脂肪肝,引发机体全身性炎症反应及主动脉血管和肝脏局部炎症反应。中药活血方和阳和汤通过不同机制、不同程度地抑制炎症反应,以独立于降脂作用的方式,调节高脂血症引发全身炎症反应、主动脉血管和肝脏局部免疫反应,抑制或延缓AS和NAFLD的发生发展。
Objective： To observe and compare the effect of Huoxue-Fang and Yanghe-Decoction on AS and NAFLD of apoE-deficient mice with Western diet （WD） by modulating systemic immune responses, liver and aortic immune responses. Methods： In the present study, 10 week-old homozygous female apoE^-/- mice were fed with a normal chow diet, a high cholesterol WD and WD supplemented with Huoxue-Fang and Yanghe-Decoction. After 18 weeks, the proportions of monocyte and subtypes of monocyte, and their surface receptor TLR4 in peripheral blood were detected by flow cytometry. Serum cytokines were detected by suspension array technology. The pathogenesis of aorta was detected by HE staining, the pathogenesis liver was detected by Oil-Red O staining and cytokiues in aorta and liver were detected by real-time PCR. Results： 18-week WD increased the levels of TC and LDL in apoE^-/- mice. Hyperlipidemia increased the proportions of monocyte and Ly6C^＋＋ , and expression of their surface receptor TLR4 in apoE^ -/- mice; and induced serum IL-10, IL-1b, MCP-1, M-CSF and TNF-a levels. In aorta, 18-week WD induced atherosclerosis lesions of aorta in apoE^-/- mice, increased expressions of cytokines TNF-a, MCP-1, IFN-γ, NOS2 and IL-lb expression. In liver, 18-week WD induced NAFLD in apoE^-/- mice, increased expressions of cytokines TNF-α, IL-6, MCP-1, IFN-γ, NOS2 and IL-lb expression. After 18 weeks admistration of Houxue-Fang, Yanghe-Decoction on ApoE -/- mice with WD, these medicines did not change the serum lipid levels significantly. Houxue-Fang decreased the proportions of monocyte and Ly6C^＋＋. Huoxue-Fang and Yanghe-Decoction decreased TLR4expression in monocyte and Ly6C^＋＋ Huoxue-Fang lowered serum IL- 10, IL-lb, MCP-1, TNF-a and M-CSF level, and Yanghe-Decoction just decreased TNF-α level. In liver, Huoxue-Fang and Yanghe-Decoction decreased steatosis. Huoxue-Fang and Yanghe-Decoction decreased expressions of cytokines TNF-α, MCP-1, IFN-γ, NOS2 and IL-1b in aorta and liver. And Huoxue-Fang showed more effect than Yanghe-Decoction on inhibiting steatosis and inflammatory cytokines in aorta and liver. Conclusion： Hyperlipidemia induced systemic immune responses, liver and aortic inflammatory disproportions in apoE^-/-mice. Huoxue-Fang and Yanghe-Decoction can modulate these inflammatory disproportions by different degrees and in a lipids-lower-independent manner to inhibit or delay the development of AS and NAFLD.
出处
《中国中医基础医学杂志》
CSCD
北大核心
2017年第12期1705-1709,1735,共6页
JOURNAL OF BASIC CHINESE MEDICINE
基金
国家重点基础研究发展计划(973)(2014CB542903)-中医证候临床辨证的基础研究
中国中医科学院自主选题研究项目(YZ-1407
YZ-1702)-祛痰
化瘀及祛痰化瘀对ApoE-/-动脉粥样硬化小鼠作用机制的对比研究
基于黄连解毒汤调控肠道菌群失衡抗动脉粥样硬化的机制研究
作者简介
姜楠（1992-），女，山东人，在读硕士，从事中西医结合基础免疫研究。;指导老师;通讯作者：马雅銮（1968-），女，研究员，医学博士，硕士研究生导师，从事中医药抗炎与免疫研究，Tel：010-64089027，E-mail：yaluanma@163．com。
