大鼠血管钙化模型炎症因子及其受体的表达

目的观察血管钙化大鼠模型血清炎症因子[C反应蛋白（C-reactive protein,CRP）、白细胞介素-6（interleukin-6,IL-6）和单核细胞趋化蛋白-1（monocyte chemoattractant protein-1,MCP-1）]表达的变化,以及大鼠主动脉组织中与血清炎症因子相对应的受体表达的变化,以探讨炎症因子在血管钙化中的作用和意义。方法实验动物按电脑随机数字表法分为正常组和钙化组,每组8只,钙化组采用维生素D3（300 000 U/kg1次肌肉注射）和尼古丁（25 mg/kg溶于花生油中,早、晚各灌胃1次）诱导大鼠血管钙化模型,用Von Kossa染色检测血管钙化程度,用钙离子测试盒、碱性磷酸酶（alkaline phosphatase,ALP）试剂盒测定大鼠主动脉钙含量和ALP活性,用放射免疫法检测大鼠血清炎症因子含量,用免疫组织化学法检测血管组织炎症因子受体表达。结果Von Kossa染色可见血管钙化模型大鼠主动脉有大量黑色颗粒沉淀。钙化组血管钙含量、ALP活性明显高于正常组,差异有统计学意义[分别为（0.42±0.05）μmol/g蛋白vs.（0.23±0.02）μmol/g蛋白,P〈0.01;（170.70±14.04）U/g蛋白vs.（110.30±14.05）U/g蛋白,P〈0.01]。钙化组血清炎症因子（CRP、IL-6和MCP-1）的表达比正常组明显上调,差异有统计学意义[分别为（2.20±0.14）mg/L vs.（1.69±0.24）mg/L,P〈0.01;（182.80±4.48）ng/L vs.（153.10±4.28）ng/L,P〈0.01;（83.07±2.37）ng/L vs.（70.52±2.12）ng/L,P〈0.01]。与正常组相比,血管钙化模型大鼠主动脉组织中的炎症因子相应受体（IL-6受体和MCP-1受体）的表达亦同步上调。结论钙化大鼠血清炎症因子和主动脉相应受体表达上调,提示炎症因子参与血管钙化的发生和发展过程。

Expressions of inflammatory factors and receptors in rat models of vascular calcification

Abstract：Objectives To investigate the expressions of serum inflammatory factors [C-reactive protein （CRP）, interleukin-6 （IL-6）, monocyte chemoattractant protein （MCP-1 ）] and corresponding receptors in rat models of aorta vascular calcification, and to explore the effects and the significance of inflammatory factors in vascular calcification. Methods SD rats were randomly divided into normal group and calcification group （n=8 respectively）. The rats in calcification group received a single intramuscular injection of vitamin D3 （300 000 U/kg） and two doses of nicotine （25 mg/kg dissolved in peanut oil） gavaged in the morning and evening to induce vascular calcification. Vascular calcification was confirmed by Von Kossa staining. Calcium content and alkaline phosphatases （ALP） activity were detected by calcium assay kit and ALP detection kit respectively. Serum inflammatory factor contents were determined by radioimmunoassay, and the expressions of corresponding receptors were determined by immunohistochemistry. Results Von Kossa staining showed that there were a large number of black granules deposited in rat models of aorta vascular calcification. Calcium content and ALP activity in calcification group were significandy higher than those in normal group [ （0.42±0.05）μmol/g protein vs. （0.23±0.02） μmol/g protein, P〈0.01; （170.70±14.04） U/g protein vs. （110.30±14.05） U/g protein, P〈0.01]. Meanwhile, serum inflammatory factor （CRP, IL-6 and MCP-1 ） expressions in calcification group obviously raised compared with those in normal group [CRP： （2.20±0.14） mg/L vs. （1.69±0.24） rag/L, P〈0.01; IL-6： （182.80±4.48） ng/Lvs. （153.10±4.28） ng/L, P〈0.01; MCP-1： （83.07±2.37） ng/L vs. （70.52±2.12） ng/L, P〈0.011. Expressions of corresponding receptors （IL-6 receptor and MCP-1 receptor） in calcification group were significantly up-regulated at the same time. Conclusions Inflammatory factors and corresponding receptors are significantly up-regulated in rat models of aorta vascular calcification, which suggests that inflammatory factors may be involved in the occurrence and the development of vascular calcification.