IL-10拮抗Pg-LPS所致高脂血症兔枯否氏细胞炎症反应的实验研究  

Experimental Study of IL-10 anta Ponizing Inflammatory Reaction of Kuffer Cell Induced by Pg-LPS in Hyperlipidemia Rabbit

作　　者：李艳芬[1] 许雯静[2] 吴春芳[1] 游晓庆[1] 骆凯[1] 闫福华[3] 

机构地区：[1]福建医科大学附属口腔医院牙周科,福州350002 [2]福建省人民医院口腔科,福州350004 [3]南京大学医学院附属口腔医院,南京210008

出　　处：《福建医科大学学报》2016年第6期359-364,共6页Journal of Fujian Medical University

基　　金：国家自然科学基金(30973326);福建省自然科学基金(2013J01306);福建省教育厅科研基金(JA11123)

摘　　要：目的探讨白细胞介素-10(IL-10)对牙龈卟啉单胞菌脂多糖(Pg-LPS)引起的高脂血症兔肝脏枯否氏细胞(KC)炎症反应的影响。方法健康新西兰兔12只随机分为2组,分别喂予基础饲料和高脂饲料。喂养6周后建立高脂血症模型。分离、收集KC,将正常和高脂组KC随机分为:对照组、Pg-LPS组(1μg/mL Pg-LPS刺激)、IL-10+Pg-LPS组(0.1μg/mL IL-10+1μg/mL Pg-LPS刺激)。刺激24h后,采用Griess法检测NO,Western-blot法检测NF-κB p65及IκB-α,DCFH-DA荧光检测细胞内ROS的表达。结果对照组中高脂KC的NF-κB p65蛋白及NO,ROS水平均高于正常KC。Pg-LPS作用后,正常及高脂KC的炎症物质表达量升高,且高脂血症与Pg-LPS呈协同作用。IL-10的干预处理使NO,ROS,NF-κB p65表达下降,IκB-α表达升高,对Pg-LPS所引起的炎症反应起抑制作用,对高脂KC的抑制作用更加明显。结论高脂血症可使KC处于相对激活状态。在牙周致病菌Pg-LPS作用下,高脂血症和Pg-LPS具有协同作用,IL-10可抑制Pg-LPS所致KC的炎性反应,且对高脂状态下KC的干预作用更佳。Objective To explore the effect of interleukin-10 on inflammatory reaction of Kuffer cell following the Pg-LPS stimulation in hyperlipidemia rabbit. Methods 12 New Zealand rabbits were randomly divided into two groups and were fed with normal or high fat diet. The hyperlipidemia model was established 6weeks later. We isolated and cultured the Kuffer cells（KC）. Both normal and hyperlipidemia group were divided into 3groups：control,Pg-LPS at 1μg/mL,and Pg-LPS at 0.1μg/mL IL-10＋1μg/mL. After the treatment for 24 h,NO levels were detected by Griess assay. NF-κB p65 and IκB-αfragment were inspected by Western blot. ROS levels were determined by fluorescence enzymelabelled meter with DCFH-DA as fluorescent probe. Results The levels of NF-κB p65,NO and ROS were higher in hyperlipidemia control group than in normal control group. After being stimulated by Pg-LPS,the expression of inflammatory substances was increased indicating a synergistic effect between hyperlipidemia and Pg-LPS. Under the IL-10 treatment the expression of NO,ROS,NF-κB p65 were decreased,while the expression of IκB-αwas increased. IL-10 inhibited the inflammatory reaction induced by Pg-LPS,and the inhibition was more significant in the KC of hyperlipidemia rabbit. Conclusion The KC of hyperlipidemia rabbit is in a relatively activated condition,after being stimulated by Pg-LPS,hyperlipidemia and Pg-LPS present a synergistic effect. IL-10 can inhibit the inflammatory reaction of KC after the Pg-LPS stimulation,and the effect is better for the KC with hyperlipidemia.

关 键 词：＊卟啉单胞菌 牙髓 脂多糖类 白细胞介素10 高脂血症 炎症 

分 类 号：R589.2[医药卫生—内分泌]