巨噬细胞移动抑制因子和细胞间黏附分子-1在慢性牙周炎合并动脉粥样硬化大鼠模型中的表达分析

Expression analysis of macrophage migration inhibitory factory and intercellular cell adhesion molecule-1 in experimental rats with periodontitis and atherosclerosis

目的分析慢性牙周炎(CP)合并动脉粥样硬化(AS)大鼠血清及动脉组织中巨噬细胞移动抑制因子(MIF)以及细胞间黏附分子-1(ICAM-1)的表达。方法24只成年雄性SD大鼠用随机数字表法分为四组,分别为正常对照(NC)组、CP组、AS组及CP+AS组,每组各6只。采用钢丝结扎双侧上颌第二磨牙联合口腔内接种牙龈卟啉单胞菌(P.gingivalis)建立牙周炎模型;高脂饮食、腹腔注射维生素D3联合左侧颈动脉球囊损伤法建立AS模型。四组分别接受相应的建模处理。12周后,组织病理学观察颈动脉组织的病理改变;体视显微镜下观察各组大鼠牙槽骨吸收面积;全自动生化分析仪检测大鼠血清血脂的表达水平;酶联免疫吸附测定(ELISA)和免疫组织化学法分析血清中和主动脉组织中MIF及ICAM-1的表达水平。结果组织病理学检查发现:NC组颈动脉组织未见明显异常,CP组血管壁较NC组略增厚,AS组、CP+AS组颈主动脉组织中出现动脉粥样硬化斑块病变;AS组、CP+AS组血清三酰甘油、总胆固醇、低密度脂蛋白(LDL)水平较NC组、CP组明显升高(P<0.05),AS组、CP+AS组血清高密度脂蛋白(HDL)水平较NC组、CP组明显降低(P<0.05)。两项检测结果提示AS模型建立成功。ELISA检测血清MIF表达水平,AS组(50.88±4.20)μg/L、CP+AS组(59.40±3.92)μg/L明显高于NC组(42.93±2.63)μg/L及CP组(45.57±2.59)μg/L(P<0.05),且CP+AS组表达水平最高(P<0.05);CP组(3.99±0.44)μg/L、AS组(4.19±0.89)μg/L、CP+AS组(6.77±1.47)μg/L血清ICAM-1表达水平明显高于NC组(1.33±0.25)μg/L(P<0.05),且CP+AS组表达水平最高(P<0.05),CP与AS组间差异无统计学意义(P>0.05)。免疫组织化学法检测动脉组织中MIF表达(吸光度),AS组(0.127±0.011)、CP+AS组(0.152±0.022)明显高于NC组(0.096±0.010)、CP组(0.010±0.011)(P<0.05),且CP+AS组表达水平最高(P<0.05);AS组(0.183±0.025)、CP+AS组(0.197±0.033)动脉组织中ICAM-1表达(吸光度)明显高于NC组(0.133±0.012)、CP组(0.156±0.006)(P<0.05),AS组与CP+AS组间差异无统计学意义(P>0.05)。结论同CP组和AS组相比,CP+AS组血清及主动脉组织中MIF的表达水平及血清中ICAM-1的表达水平明显增高,提示慢性牙周炎可能通过升高炎性因子MIF及ICAM-1表达的方式促进AS病变的形成过程。

Objective To analyze the expression of macrophage migration inhibitory factor(MIF)and intercellular cell adhesion molecule-1(ICAM-1)in the serum and arterial tissue of rats with chronic periodontitis(CP)and atherosclerosis(AS).Methods Twenty-four adult male SD rats were assigned into 4 groups by random number table method:normal control group(NC),the CP group,the AS group and the CP+AS group,each group with 6 rats.CP animal models were established by wire ligation on bilateral maxillary second molars and oral inoculation with Porphyromonas gingivalis(P.gingivalis).AS animal models were induced by carotid artery balloon injury and injected vitamin D3 intraperitoneally and fed with a high-fat diet.Four groups received corresponding modeling processing.Twelve weeks later,pathological changes in carotid arteries were observed by histopathology.The alveolar bone resorption levels of rats were observed by stereomicroscope.The serum lipid levels of rats were detected through automatic biochemical analyzer.The levels of MIF and IC.AM-1 in serum were detected by Enzyme-linked immunosorbent assays(ELISAs),the protein expression levels of MIF and ICAM-1 in carotid artery tissue were detected using immunohistochemiatry.Results Histopathological examination showed that there were no obvious abnormalities in the vessel wall of the NC group,the vessel wall of the CP group were slightly thickened,and the atherosclerotic plaque lesions of the AS group and the CP+AS group were formed.The levels of serum triglyceride(TC),total cholesterol and low density lipoprotein cholesterol(LDL)in the AS group and the CP+AS group were significantly higher than those in the NC group and the CP group(P<0.05),the level of serum high density lipoprotein cholesterol(HDL)in the AS group and the CP+AS group was significantly lower than that in the NC group and the CP group(P<0.05).Two test results suggested that the AS animal model was successfully established.The expression of MIF in the serum of the AS group(50.88±4.20)p g/L and the GP+AS group(59.40±3.92)μg/L was significantly higher than that in the NC group(42.93±2.63)μg/L and the CP group(45.57±2.59)μg/L(P<0.05),MIF expression of the CP+AS group was the highest(P<0.05).The expression of serum ICAM-1 in the CP group(3.99±0.44)μg/L,the AS group(4.19±0.89)μg/L and the CP+AS group(6.77±1.47)μg/L was significantly higher than that in the NC group(P<0.05),and the expression of the CP+AS group was the highest(P<0.05),and there was no significant difference between the CP group and the AS group(P>0.05).Immunohistochemistry showed that the expression of MIF in the carotid artery tissue of the AS group and the CP+AS group was significantly higher than that of the NC group and the CP group(P<0.05),and the expression of the CP+AS group was the highest(P<0.05).The expression of ICAM-1 in arterial tissues of the AS group and the CP+AS group was significantly higher than that of the NC group and the CP group(P<0.05).Conclusion The expression of MIF in the serum and carotid artery tissue and the expression of ICAM-1 in the serum are significantly increased compared with the CP group and the AS group,suggesting that chronic periodontitis may promote the development of atherosclerotic lesions by increasing the expression levels of MIF and ICAM-1.