摘要
目的观察生地黄水煎总提物对动脉粥样硬化(AS) Apo E-/-小鼠脂质斑块的改善作用以及对"IL-34"-Rho/Rock通路的影响。方法①动物实验:将40只Apo E-/-小鼠分成空白对照组(CTL组)、模型对照组(Model组)、5μg·g^(-1)生地黄干预组(Reh-L组)和10μg·g^(-1)生地黄干预组(Reh-H组)。空白对照组小鼠给予普通饲料,其余组小鼠腹腔注射人血清低密度脂蛋白(LDL) 2周,并喂以高脂饲料。连续灌胃4周,检测小鼠血脂指标,并计算AS指数(AI)。②细胞实验:将RAW264. 7细胞分为空白对照组(con组)、ox-LDL组、IL-34组和Reh组;油红O染色观察泡沫细胞形成,检测细胞内胆固醇酯含量和胆固醇流出率。免疫荧光染色法、qRT-PCR法和Western blot法检测Rho A和Rock1表达量。结果①动物实验:Reh-H组小鼠外周血TC、TG、LDL-C、内皮素-1(ET-1)、白介素-34(IL-34)、单核细胞趋化蛋白1(MCP-1)、巨噬细胞炎性蛋白2(MIP-2)水平、AI值以及AS斑块面积比低于Model组和Reh-L组,而HDL-C水平高于Model组和Reh-L组(P <0. 05)。经qRT-PCR法和Western blot法检测,Reh-H组小鼠主动脉组织Rho A和Rock1表达量较Model组和Reh-L组降低(P <0. 05)。②细胞实验:经oxLDL和IL-34共诱导,RAW264. 7细胞内胆固醇酯含量、胆固醇流出率以及Rho A和Rock1蛋白表达量较con组和ox-LDL组细胞增加(P <0. 05)。而加入Reh干预后泡沫细胞数量、胆固醇酯含量、胆固醇流出率以及RAW264. 7细胞Rho A和Rock1蛋白表达量较IL-34组细胞明显减少(P <0. 05)。结论生地黄可通过抑制"IL-34"-Rho/Rock信号通路抑制巨噬细胞吞噬脂质形成泡沫细胞,从而有效地预防AS的形成和进展。
OBJECTIVE To investigate the protective effects and mechanism of extracts of Rehmanniae against atherosclerosis by'IL-34'-Rho/Rock pathway in Apo E-/-mice.METHODS①Animal experiment:40 Apo E-/-mice were randomly divided into CTL group,Model group,Reh-L group(5μg·g-1)and Ren-H group(10μg·g-1),which were induced by low-density lipoprotein(LDL)for AS models.The serum levels of interleukin-34(IL-34),LDL,mouse monocyte chemotactic protein 1(MCP-1),mouse macrophage inflammatory protein 2(MIP-2)and endothelin-1(ET-1)were detected by ELISA.AS index(AI)was calculated according to the serum lipids levels.②Macrophage experiment:RAW264.7 macrophages were cultured in vitro and divided into con group,ox-LDL group,IL-34 group and Reh group.IL-34-indeced foam cells formation by oil red O staining.The cholesterol ester levels and cholesterol efflux of RAW264.7 macrophages were detected.The immunofluorescence,qRT-PCR,Western blot analysis were used to detect Rho A and Rock1 expression.RESULTS①Animal experiment:the serum levels of TC,TG,LDL-C,ET-1,IL-34,MCP-1,MIP-2 and AI,atherosclerotic lesion areas of the thoracic aorta of mice in Reh-H group were lower than Model group and Reh-L group,but with higher HDL-C level(P<0.05).Compared with the CTL group,the Rho A and Rock1 levels of aorta tissues in Model group were significantly increased by qRT-PCR and Western blot(P<0.05).But the Rho A and Rock1 levels of aorta tissues in Reh-H group were lower than Model group or Reh-L group(P<0.05).②Macrophage experiment:the cholesterol ester level and cholesterol efflux,Rho A and Rock1 expressions of RAW264.7 macrophages induced by ox-LDL and IL-34 were more than con group and ox-LDL group(P<0.05).Compared with IL-34 group,the cholesterol ester level and cholesterol efflux,Rho A and Rock1 expressions of RAW264.7 macrophages in Reh group were significantly decreased(P<0.05).CONCLUSION The extracts of Rehmanniae can protectagainst atherosclerosis in Apo E-/-AS mice models induced by LDL through inhibiting'IL-34'-Rho/Rock pathway.
作者
刘艳宾
秦洁洁
杨树涵
陈红伟
邢永生
张文才
LIU Yan-bin;QIN Jie-jie;YANG Shu-han;CHEN Hong-wei;XING Yong-sheng;ZHANG Wen-cai(Department of Internal Medicine,Xinxiang Central Hospital;Coronary Care Unit,Xinxiang Central Hospital,Henan Province,Xinxiang 453000,China;Department of Cardiology,First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中国药学杂志》
CAS
CSCD
北大核心
2019年第8期624-631,共8页
Chinese Pharmaceutical Journal
基金
国家自然科学基金项目资助(81500362)
