摘要
目的:探讨异甘草素通过激活PPAR⁃γ信号通路调控ox⁃LDL代谢稳定动脉粥样硬化(AS)斑块的分子机制。方法:采用高脂喂养+右侧颈总动脉外置套管术(PCCP)制备ApoE⁃/⁃小鼠AS颈动脉斑块模型。ApoE⁃/⁃小鼠经PCCP术后随机分为模型组和异甘草素组,正常组采用C57BL/6J小鼠。术后高脂饲料继续喂养8周,建立AS模型。全自动生化仪检测血清中总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL⁃C)、高密度脂蛋白胆固醇(HDL⁃C)含量。ELISA检测血清中氧化低密度脂蛋白(ox⁃LDL)含量。HE染色观察颈动脉病理形态,并测定颈动脉参数。油红O染色用于脂质测定,Masson染色用于胶原含量测定,MOMA⁃2和α⁃SMA免疫组织化学染色用于巨噬细胞和平滑肌细胞测定,并计算易损指数。Western blot检测小鼠动脉中PPAR⁃γ、LXR⁃α、FABP⁃4、MMP⁃2及MMP⁃9蛋白表达。结果:与正常组相比,模型组TC、TG、LDL⁃C、HDL⁃C和ox⁃LDL升高。与模型组相比,异甘草素组TC、TG、LDL⁃C和ox⁃LDL降低,HDL⁃C无明显变化。与正常组相比,模型组小鼠颈动脉的内膜厚度(IT)、内膜/中膜厚度(IT/MT)、斑块面积(PA)、斑块面积/血管管腔面积(PA/LA)均增加,斑块内脂质和MOMA⁃2含量增加,胶原和α⁃SMA含量减少,易损指数较高,PPAR⁃γ、LXR⁃α表达减少,FABP⁃4、MMP⁃2、MMP⁃9表达增加。与模型组比较,异甘草素组小鼠颈动脉IT、IT/MT、PA、PA/LA均减少,斑块内脂质和MOMA⁃2含量减少,胶原和α⁃SMA含量增加,易损指数降低,PPAR⁃γ、LXR⁃α表达增加,FABP⁃4、MMP⁃2、MMP⁃9表达减少。结论:异甘草素能够通过激活PPAR⁃γ、上调LXR⁃α,减少FABP⁃4表达,降低ox⁃LDL水平,减少MMP⁃2和MMP⁃9的蛋白表达,降低斑块易损指数,增加斑块稳定性,发挥抗AS的作用。
Objective:To explore the molecular mechanisms of isoliquiritigenin in stabilizing atherosclerotic plaques by acti⁃vating PPAR⁃γsignal pathway to regulate ox⁃LDL metabolism.Methods:The ApoE⁃/⁃mice AS carotid plaque model was prepared by using high fat diet and right perivascular carotid collar placement(PCCP).ApoE⁃/⁃mice were randomly divided into the model group and the isoliquiritigenin group after PCCP.C57BL/6J mice were used for the normal group.High fat diet continued feeding for 8 weeks after PCCP to establish the AS model.Automatic biochemical analyzer was used to test levels of total cholesterol(TC),tri⁃acylglyceride(TG),low⁃density lipoprotein cholesterol(LDL⁃C)and high⁃density lipoprotein cholesterol(HDL⁃C).ELISA was used to measure oxidized low⁃density lipoprotein(ox⁃LDL)in serum.Hematoxylin⁃eosin(HE)staining was used to observe the pathological pattern of the carotid artery,and then the carotid parameters were calculated.Oil red O staining was used for lipid deter⁃mination,Masson staining was used to determine collagen content,MOMA⁃2 andα⁃SMA immunohistochemical staining were used to determine macrophages and smooth muscle cells,and to calculate the vulnerability index.Western blot was used to detect the ex⁃pression of PPAR⁃γ,LXR⁃α,FABP⁃4,MMP⁃2 and MMP⁃9 in mice arteries.Results:Compared with the normal group,TC、TG、LDL⁃C、HDL⁃C and ox⁃LDL were increased in the model group.Compared with the model group,TC,TG,LDL⁃C and ox⁃LDL were reduced,and there was no significant change in HDL⁃C of the isoliquiritigenin group.Compared with the normal group,intima thickness(IT),intima/media thickness(IT/MT),plaque area(PA),and plaque area/lumen area(PA/LA)of carotid arteries were increased,the content of lipid and MOMA⁃2 in plaques was increased,collagen andα⁃SMA content decreased,and the vulner⁃ability index was higher in the model group.The expressions of PPAR⁃γand LXR⁃αwere reduced and the expressions of FABP⁃4,MMP⁃2 and MMP⁃9 were increased in the model group.Compared with the model group,carotid IT,IT/MT,PA,and PA/LA were reduced,the content of lipid and MOMA⁃2 in plaques was decreased,collagen andα⁃SMA content were increased,and the vul⁃nerability index was decreased in the isoliquiritigenin group.PPAR⁃γand LXR⁃αexpression were increased,FABP⁃4,MMP⁃2 and MMP⁃9 expression were decreased significantly in the isoliquiritigenin group.Conclusion:Isoliquiritigenin can exert anti⁃AS effects by activating PPAR⁃γ,up⁃regulating LXR⁃α,reducing FABP⁃4 expression,reducing ox⁃LDL,reducing the protein expression of MMP⁃2 and MMP⁃9,decreasing plaque vulnerability index,and increasing plaque stability.
作者
许心蕊
高照
张晴玥
杨漫芳
孙浩
冯露
王添钰
李洋
娄利霞
吴爱明
聂波
XU Xin-rui;GAO Zhao;ZHANG Qing-yue;YANG Man-fang;SUN Hao;FENG Lu;WANG Tian-yu;LI Yang;LOU Li-xia;WU Ai-ming;NIE Bo(Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing,Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100700,China;Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Department of Pathology,Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine,Beijing 100700,China)
出处
《海南医学院学报》
2023年第18期1367-1374,共8页
Journal of Hainan Medical University
基金
国家自然科学基金(82274488,81874446)。
