摘要
目的 探讨线粒体靶向多肽SS-31对载脂蛋白E基因敲除型[ApoE(-/-)]小鼠动脉粥样硬化的影响.方法 雄性8周ApoE(-/-)小鼠30只,均分为对照组和药物组.药物组小鼠皮下注射SS-31(1 mg·kg-1·d-1),对照组皮下注射等量0.9％生理盐水,高脂喂养12周后处死.主动脉大体油红染色评估斑块面积(纵切面),主动脉窦区油红染色评估斑块大小(横切面).主动脉窦区二氢乙啶(DHE)染色及8-羟基脱氧鸟苷(8-OHdG)免疫组化反应氧化应激水平,试剂盒检测主动脉三磷酸腺苷含量.CD68、α-SMA免疫组化及Masson染色反应斑块组分变化,Western blotting检测主动脉超氧化物歧化酶(SOD)2蛋白水平,试剂盒检测SOD活性.主动脉窦区CD36、脂氧合酶(LOX)-1和三磷酸腺苷结合盒转运体A1免疫组化评估脂质代谢情况.酶联免疫吸附试验法检测小鼠血清中细胞内黏附分子-1(ICAM-1)、单核细胞趋化因子-1(MCP-1)、白细胞介素6(IL-6)和C反应蛋白等炎症因子水平.结果 药物组小鼠主动脉斑块面积和大小减少,斑块处巨噬细胞减少,平滑肌细胞增多,胶原增多.药物组小鼠主动脉切片DHE荧光减弱,8-OHdG表达上调,SOD酶活增加.药物组小鼠血清ICAM-1、MCP-1、IL-6水平下降,药物组小鼠主动脉脂质摄取蛋白CD36、LOX-1表达下降.结论 药物SS-31可以减轻ApoE(-/-)小鼠动脉粥样硬化,为动脉粥样硬化治疗提供新思路.
Objective To investigate whether Szeto-Schiller-31(SS-31) could suppress the development of atherosclerosis in vivo. Methods Thirty male ApoE (-/-) mice (8 weeks old) fed with western diet were treated with normal saline or SS-31 (1 mg·kg-1·d-1) (15 mice for each group) through subcutaneous injection for 12 weeks. Oil Red O staining was performed to evaluate area and sizes of the plaques. DHE staining and immunohistochemical staining of 8-hydroxy-deoxyguanosine (8-OHdG) was performed to assess the oxidative stress. The aorta ATP contents were assessed by the ATP bioluminescence assay kit. Immunohistochemical staining of CD68 and α-SMA and Masson's trichrome staining were performed to evaluate the composition of atherosclerotic plaque. Biochemical assays were performed to determine the protein level and activity of superoxide dismutase (SOD). The levels of CD36, lipoxygenase-1 (LOX-1) and ATP-binding cassette transporter A1 were immunohistochemically determined to evaluate the cholesterol transport in aorta. Inflammatory factors, including intracellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) and C-reactive protein (CRP) in serum, were detected through ELISA. Results SS-31 administration reduced the area and sizes of western diet-induced atherosclerotic plaques and changed the composition of the plaques in ApoE (-/-) mice. Oxidative stress was suppressed, as evidenced by the reduced DHE stain, down -regulated 8 -OHdG expression, and increased SOD activity after chronic SS -31 administration. Moreover, systemic inflammation was ameliorated as seen by decreased serum ICAM-1, MCP-1, and IL-6 levels. Most importantly, SS-31 administration inhibited cholesterol influx by down-regulating expression of CD36 and LOX-1 in aorta to prevent lipid accumulation to further suppress the atherosclerotic progression. Conclusion Administration of SS-31 prevents against atherosclerotic formation in ApoE (-/-) mice suggesting that SS31 might be considered to be a potential drug to prevent atherosclerotic progression.
出处
《中华血管外科杂志》
2017年第4期223-229,共7页
Chinese Journal of Vascular Surgery
基金
国家自然科学基金面上项目(81370387)Fund program: National Natural Science Foundation of China(General Program)
关键词
动脉粥样硬化
氧化应激
炎症
脂质
Atherosclerosis
Oxidative stress
Inflammation
Lipid
