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目的:研究ADRP与As发生和发展的关系。方法:使用80mg/LOx -LDL和/或1mmol/L反义ADRP寡核苷酸与小鼠腹腔巨噬细胞共孵育72h。测定细胞内胆固醇酯;油红O染色显示细胞内中性脂质;RT -PCR和Westernblotting观察反义寡核苷酸对ADRP表达的影响。高胆固醇饲料喂养新西兰白兔12周,复制As模型。测定血清总胆固醇、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、甘油三酯及动脉壁胆固醇;苏丹Ⅳ染色显示主动脉病变面积;HE染色观察主动脉及肝脏的病变;免疫组织化学方法显示ADRP在主动脉病变区及肝脏中的表达。结果:反义ADRP寡核苷酸使细胞内胆固醇酯由(46. 6±3 .4 )mg/gprotein下降到(19. 9±1 .9)mg/gprotein ,细胞内中性脂质明显减少,ADRP基因和蛋白的表达显著下降。喂高胆固醇饲料的动物血清TC、LDL -C、TG及动脉壁胆固醇显著升高,主动脉病变面积为(40 . 1±7 .3) % ,ADRP在主动脉病变区免疫组织化学染色强阳性,在肝脏中染色阴性。结论:反义ADRP寡核苷酸能够明显抑制由氧化低密度脂蛋白引起的小鼠腹腔巨噬细胞中胆固醇酯的聚集。ADRP在兔As病变中表达明显增高。提示高表达ADRP促使As发生和发展。
Objective: To study the relationship between the occurrence and development of ADRP and As. METHODS: Mice peritoneal macrophages were co-incubated with 80 mg / L Ox-LDL and / or 1 mmol / L antisense ADRP oligonucleotides for 72 h. The intracellular cholesterol esters were determined by oil red O staining. The neutral lipid was detected by RT-PCR and Western blotting. The effect of antisense oligonucleotide on the expression of ADRP was observed. New Zealand white rabbits were fed with high cholesterol diet for 12 weeks to copy As model. Serum total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, triglyceride and arterial wall cholesterol were measured. The area of aortic lesions was observed by Sudan Ⅳ staining. The aorta and liver lesions were observed by HE staining. Immunohistochemistry showed that ADRP In aortic lesions and liver expression. RESULTS: Antisense ADRP oligonucleotides decreased intracellular cholesterol esters from (46.6 ± 3.4) mg / gprotein to (19.9 ± 1.9) mg / gprotein, and the intracellular free lipids decreased significantly , ADRP gene and protein expression decreased significantly. Serum levels of TC, LDL-C, TG and arterial wall cholesterol of rats fed with high-cholesterol diet were significantly increased, and the area of aortic lesions was (40.1 ± 7.3)%. Immunohistochemical staining of ADRP in aortic lesions was strongly positive , Staining negative in the liver. CONCLUSION: Antisense ADRP oligodeoxynucleotides significantly inhibit cholesterol ester accumulation in mouse peritoneal macrophages induced by oxidized low-density lipoprotein. ADRP expression in rabbit As lesions was significantly higher. Prompted the high expression of ADRP promote As occurrence and development.