水蛭对载脂蛋白E基因敲除鼠动脉粥样硬化斑块的影响

目的:探讨水蛭对载脂蛋白E基因敲除(apoE-/-)小鼠动脉粥样硬化(As)斑块的影响及可能机制。方法:(21±3)g雌性apoE-/-小鼠20只,12-13周龄,高脂喂养8周后随机分为对照组和水蛭用药组,第10周后改为普通饲养,对照组给予0.5%羧甲基纤维素钠(CMC-Na)悬液灌胃,用药组于混悬液中加入水蛭粉[0.2 mg/(g.d)]灌胃8周。小鼠禁食过夜后采取眼球后静脉血,测量血浆总胆固醇(TC)、甘油三脂(TG)水平,肿瘤坏死因子α(TNF-α)和对氧磷酯酶1(PON1)酶活性,处死小鼠后剖面分析胸腹主动脉As斑块面积,主动脉根部组织冰冻切片分析斑块面积、脂质含量以及血管平滑肌细胞(SMC)面积。结果:与对照组比较,水蛭组小鼠血浆TC及TNF-α水平明显下降,而TG水平和PON1酶活性并无显著改变;水蛭能明显减少主动脉As斑块面积,斑块中脂质含量减少但所占斑块比例无差异,而斑块中SMC比例显著减少。结论:水蛭粉喂食可能通过降低apoE-/-小鼠的血脂及炎症水平,进一步抑制As斑块中SMC增殖而发挥抗As的功能。

Effects of Leech on Atherosclerotic Lesion in Apolipoprotein E Deficient Mice

Objective: To investigate the effects of leech powder on the development of atherosclerosis lesions in apolipoprotein E deficient(apoE-/-) mice.Methods: Twenty female apoE-/-mice fed with Western diet 8 weeks,then randomly divided into control group and leech treatment group.The diets were changed to chow diet in the tenth week.Leech powder(0.2 mg/g per day) was given to hyperlipidemic mice for 8 weeks and 0.5% CMC-Na suspension was given to the control.Plasma total cholesterol(TC) and triglycerides(TG) level were determined enzymatically,tumor necrosis factor α(TNF-α) level was determined by enzyme-linked immunosorbent assay(ELISA),paraoxonase 1(PON1) activity was measured using paraoxon as the substrate.The atherosclerotic lesions in aorta and aortic root were analyzed by hematoxylin-eosin staining.The contents of lipid and smooth muscle cells(SMC) area in lesions were determined by oil-red O staining and immunohistochemistry.Results: Compared with control group,the levels of TC and TNF-α were statistically decreased,while the level of TG and PON1 activity had no obvious difference in apoE-/-mice with Leech treatment.The atherosclerotic lesions area in aorta and aortic root significantly lower after treated with leech,and the SMC content in lesions were obviously reduced while the percentage of lipid content in lesions had no changes.Conclusion: The anti-atherogenic effect of leech powder may be involved in the decrease of TC and inflammatory factor,further inhibits the proliferation of SMC in the atherosclerotic lesions.