糖尿病合并动脉粥样硬化大鼠冠状动脉巨噬细胞M1/M2及其相关指标变化

Changes of macrophages M1/M2 in coronary artery and relevant indexes in rats with diabetes mellitus and atherosclerosis

目的检测糖尿病(DM)合并动脉粥样硬化(AS)模型大鼠冠状动脉中巨噬细胞M1、M2以及冠状动脉粥样硬化病理变化,探讨其在DM合并冠状动脉粥样硬化发生、发展中的相互作用。方法将30只6周龄体质量160 g雄性Wistar大鼠正常喂养2周后,随机数字表法分成2组:实验组20只,健康对照组10只。2组均自由进水,健康对照组给予标准鼠饲料喂养。实验组行DM合并AS造模,成功造模继续高脂饲喂12周后,2组大鼠禁食12 h后处死,腹主动脉取血,离心,取血清,全自动生化分析仪检测血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C),血糖;酶联免疫吸附试验(ELISA)测定白细胞介素(IL)-6、IL-10,肿瘤坏死因子(TNF)-α;取主动脉切片苏木精-伊红(HE)染色行病理检查,并用免疫组织化学法标记主动脉巨噬细胞标志物CD86、CD163。结果实验组处死后测定TC、TG、LDL-C、HDL-C含量较健康对照组升高显著(P<0.05);实验组血清IL-6、IL-10,TNF-α较健康对照组显著升高(P<0.05),病理切片粥样硬化变化实验组明显重于健康对照组。行免疫组织化学标记冠状动脉中巨噬细胞CD86、CD163表达明显高于健康对照组。结论高血糖、高血脂及冠状动脉中巨噬细胞CD86与AS发生关系密切,可能因相互协同、诱导,一起参与AS发生、发展过程,HDL-C、IL-10可抑制巨噬细胞产生炎性因子,促进组织修复影响粥样硬化生成,但并不意味着冠心病患病风险降低,深入研究它们的共同调控机制和药物靶向定位,将可能进一步更好地控制这类疾病的发生和发展。

Objective To determine the pathological changes of macrophages M1/M2 in coronary artery and atherosclerosis of coronary artery in rat models with diabetes mellitus（DM） and atherosclerosis（AS）, and to investigate the interaction in the occurrence and development of DM and AS of coronary artery. Methods Thirty 6-week-old male Wistar rats weighing 160 g were normal fed for 2 weeks, and were randomly divided into two groups, the experimental group（n=20） and the normal control group（n=10）. Both groups could drink freely. The normal control group was fed with standard fodder. After the DM and AS models were successfully established in the experimental group, the rats were continued fed with high fat fodder for 12 weeks. After fasting for 12 h, the rats were executed. The serum was sep-arated from abdominal aortic blood by centrifugation. The serum total cholesterol（TC）, triglyceride（TG）, high density lipoprotein cholesterol（HDL-C）, low density lipoprotein cholesterol（LDL-C） and glucose were measured by the automatic biochemical analyzer. ELISA method was used to determine IL-6, IL-10 and TNF-α. Coronary artery slices were pathologically examined by HE staining. The aortic macrophage markers CD86 and CD163 were labeled by immuno-histochemistry. Results After the experimental group was executed, the determined content of TC, TG, HDL-C and LDL-C in the experimental group were significantly increased compared with those in the control group（ P〈0.05）. The levels of IL-6, IL-10 and TNF-α in the experimental group were significantly increased compared with those in the control group（P〈0.05）. The pathological changes of the AS lesions in the experimental group were more than those in the control group. The expression of macrophages CD86 and CD163 in coronary artery labeled by immunohistochem-istry in the experimental group were significantly higher than those in the control group. Conclusion Hyperglycemia,hyperlipidemia and macrophage CD86 in coronary artery is closely correlated with AS, which may be involved in the occurrence and development of AS due to mutual interaction and induction. HDL-C and IL-10 may inhibit inflammatory cytokines produced by macrophages, and promote tissue repair on AS generation. However, it does not mean that the risk of coronary heart disease is reduced. Further research on their common regulatory mechanism and drug targeting position may further control the occurrence and development of these diseases.