胆木抗apoE-/-小鼠动脉粥样硬化和非酒精性脂肪性肝病的研究

Study of Danmu on atherosclerosis and non-alcoholic fatty liver disease in apoE-/- mice

目的:观察胆木对高脂饮食导致的apoE-/-小鼠全身、主动脉血管和肝脏局部免疫反应的影响。方法:采用10周龄雌性C57BL/6和apoE-/-小鼠,分为C57BL/6+CD、apoE-/-+CD、apoE-/-+WD、apoE-/-+WD+DM组,每组16只,监测体质量变化。6周后,检测血脂和肝功能;悬液芯片检测血浆细胞因子水平;流式细胞仪检测外周血粒细胞、单核细胞及其Ly6C++亚型比例;实时定量PCR检测主动脉和肝脏细胞因子表达。干预18周后,检测主动脉和肝脏病理变化。结果:高脂6周,导致apoE-/-+WD组小鼠外周血TC和LDL、单核细胞及其炎症亚型Ly6C++比例、血浆细胞因子IL-6、TNF-α、IL-10显著升高(P<0.05,P<0.01);主动脉细胞因子IL-1β、IL-6、MCP-1、IFN-γ、TNF-α、IL-10和肝脏IL-1β、IFN-γ和IL-10表达升高(P<0.05,P<0.01)。胆木干预明显减缓小鼠体质量增长,但血脂和肝功能没有明显改变;胆木显著降低外周血单核细胞比例、血浆M-CSF和IL-10水平、主动脉IL-1β、IL-6、MCP-1、IFN-γ、TNF-α和IL-10表达和肝脏IL-1β、MCP-1、TNF-α和IL-10表达(P<0.05,P<0.01)。高脂18周导致apoE-/-小鼠主动脉AS斑块数量增多,面积增大;肝脏油红O染色阳性;胆木干预减少主动脉AS斑块面积,肝脏油红O染色减弱,并呈现由外周到血管向心性减弱。结论:高脂饮食引发apoE-/-小鼠全身性炎性反应、血管和肝脏局部炎性反应。胆木以独立于降脂作用的方式调节高脂血症引发的免疫反应,抑制动脉粥样硬化和非酒精性脂肪性肝病形成和发展。

Objective:To observe the effect of Danmu on apoE-/-mice with western diet(WD)by modulating systemic immune responses,aortic and liver immune responses.Methods:In the present study,10 weeks male C57 BL/6 and apoE-/-mice were fed,the mice were divided into C57 BL/6+CD,apoE-/-+CD,apoE-/-+WD,apoE-/-+WD+DM group,16 mice each group,the body mass change was observed.After 6 weeks,serum lipid levels by routine method,plasma cytokine levels were measured and suspension microarray.The proportions of granulocyte and monocyte and Ly6 C++were detected by flow cytometry.The expression of cytokines in aorta and liver was detected by real-time quantitative PCR.After 18 weeks of intervention,the pathological changes of aorta and liver were detected.Results:After 6-week’s WD,the levels of TC and LDL in apoE-/-+WD group were increased.Hyperlipidemia increased the proportion of monocytes,their inflammatory subtypes Ly6 C++and the levels of cytokines IL-6,TNF-αand IL-10 in plasma(P<0.05,P<0.01).6-week’s WD induced the expression of cytokines IL-1β,IL-6,MCP-1,IFN-γ,TNF-α,IL-10 in aorta and IL-1β,IFN-γ,IL-10 in liver tissue(P<0.05,P<0.01).After 6 weeks of intervention by Danmu,the weight gain of mice was significantly slowed down,but the blood lipid level and the liver function were not affected.Danmu significantly reduced the proportion of monocytes,decreased plasma cytokine M-CSF and IL-10 levels.Danmu significantly lowered the expression of cytokines IL-1β,IL-6,MCP-1,IFN-γ,TNF-α,IL-10 in aorta and IL-1β,MCP-1,TNF-α,IL-10 in liver(P<0.05,P<0.01).After 18 weeks of intervention,WD increased the number and area of aortic plaques in apoE-/-mice;liver oil red O staining was positive.Danmu reduced the area of aortic AS plaques,liver oil-red O staining was weakened,and showed diffuse attenuation from the periphery to the vascular center.Conclusion:WD induces systemic inflammatory response,aorta and liver local inflammatory responses in apoE-/-mice.Danmu regulates these inflammatory disproportions induced by hyperlipidemia,and inhibits the formation and development of atherosclerosis and non-alcoholic fatty liver disease.