高糖高脂喂养LDLR^(-/-)小鼠动脉钙化的研究

High-sugar,high-fat diet induced aortic calcification in low density lipoprotein receptor gene deficient mice

目的:研究高糖、高脂喂养LDL受体基因缺陷(LDLR-/-)小鼠的代谢及血管钙化指标,探讨糖尿病血管钙化的相关机制。方法:高糖、高脂喂养LDLR-/-小鼠为实验组(HF组),常规饲养LDLR-/-小鼠为对照组(CON组),喂养16周。实验结束检测两组小鼠空腹血糖(FBS)、总胆固醇(CHO)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、空腹胰岛素(FIns)。流动注射分析法检测血清糖基化终产物肽(AGE-P),实时定量PCR分析小鼠主动脉MSX2、OPN、Cbfα-1、α-SMA mRNA表达,苏木精-伊红染色法(HE染色)观察主动脉粥样硬化病变,Von Kossa染色观察主动脉钙化病变。结果:与CON组比较,HF组小鼠血清FBS、FIns、稳态胰岛素评价指数、CHO、TG、LDL、HDL、AGE-P水平显著增高;HF组小鼠主动脉MSX2、OPN、Cbfα-1基因表达增加,α-SMA基因表达减少。光镜下观察,HE染色CON组主动脉未见动脉粥样硬化病变,HF组主动脉可见动脉粥样硬化。Von Kossa染色CON组主动脉未见钙化,HF组主动脉可见钙化。结论:高糖、高脂喂养可以加重LDLR-/-小鼠糖、脂代谢异常,继而加速动脉钙化。这一动物模型可用于糖尿病相关血管钙化的研究。

Objective： To explore the mechanisms of diabetic vascular calcification by studying the indicators of metabolism and vascular calcification in the high-sugar,high-fat diet low density lipoprotein receptor gene deficient（ LDLR- /-） mice. Methods： High-sugar,high-fat diet fed LDLR- /-mice was the experimental group（ HF group） and conventional diet fed LDLR- /-mice was the control group（ CON group）. The two groups were fed for16 weeks. At the end of the experiment,fasting blood glucose（ FBS）,total cholesterol（ CHO）,triglyceride（ TG）,low density lipoprotein（ LDL）,high density lipoprotein（ HDL）,fasting insulin（ FIns） were detected.Serum advanced glycation end product-peptide（ AGE-P） were detected by flow injection analysis. The expressions of MSX2,OPN,Cbfα-1,α-SMA mRNA were analyzed by Real-time quantitative PCR. Aortic atherosclerotic lesions were observed by hematoxylin-eosin staining. Aortic calcified lesions were observed by Von Kossa staining.Results： Compared with CON group,the serum FBS,FIns,HOMA-IR,CHO,TG,LDL,HDL,AGE-P were significantly higher in HF group. The gene expressions of MSX2,OPN,Cbfα-1 increased,while α-SMA decreased. Aortic atherosclerotic lesions were seen in HF group. HF group demonstrated aortic calcification by Von Kossa staining. No aortic calcification was showed in the CON group. Conclusion： The high sugar,high fat diet can increase the metabolism disorder of glucose and lipid in the LDLR- /-mice,and then accelerate aortic calcification. This animal model can be used to study the mechanisms of vascular complications associated with diabetes.