摘要
目的本研究以胶原诱导性关节炎（CIA）大鼠为研究对象，通过比较丙酮酸乙酯干预前后正常大鼠、CIA大鼠及与丙酮酸乙酯（EP）干预组大鼠关节滑膜组织高迁移率族蛋白1（HMGBl）的表达，阐明HMGBI在关节炎中的作用及EP对其拮抗作用。方法实验分为正常对照组、CIA大鼠组与EP干预组，每组12只大鼠，分别在造模开始的第6、9周每组解剖6只大鼠。采用免疫组织化学检测关节滑膜组织HMGBl的表达，以病理图像分析软件IPP对胞质部位进行图像分析，表达水平以表达强度表示；采用实时定量．聚合酶链反应（PCR）检测各组大鼠滑膜组织HMGBlmRNA的表达量，以正常对照组第6周大鼠滑膜组织HMGB1mRNA表达量为参照的相对定量来表达。采用t检验比较各组间大鼠滑膜组织HMGBl的表达。结果关节滑膜组织HMGBl免疫组织化学检测结果：第6周时，正常对照组、CIA大鼠组及EP干预组表达强度分别为2．1±0．6、7．3±1．2、6．0±1．2；第9周时，分别为2．2±0．7、12．4±4．5、5．5±1．0；大鼠滑膜组织HMGBlmRNA实时定量PCR检测结果：第6周时，正常对照组、CIA大鼠组及EP干预组相对定量分别为1、2．865、2．602；第9周时，分别为1．005、4．694、1．729。在第6、9周采用免疫组织化学及实时定量PCR检测HMGBl在CIA组的表达明显高于正常对照组（P〈0．05），CIA组关节滑膜中HMGBl的表达在第9周时较第6周明显增高（P〈0．05）；第6周时，EP干预组滑膜HMGBl的表达较CIA组降低，但差异无统计学意义（P〉O．05），第9周时，其表达明显降低（P〈0．05）。结论HMGBl在CIA大鼠滑膜中的表达量明显增高，在晚期尤甚，推测HMGBl可能作为炎症因子参与了CIA的疾病过程，尤其作为晚期致炎因子发挥重要的作用；经EP干预后HMGBl表达量明显下降，推测EP可能对HMGBl具有拮抗作用，为RA患者提供新的的治疗手段。
Objective In this study, we elucidated the role of high mobility group box chromosomal protein 1 （HMGB1） in collagen-induced arthritis （CIA） rat and the antagonist role of ethyl pyruvate by using a rat model of CIA as the research object by comparing the expression of HMGB1 in normal control group, CIA model group and ethyl pyruvate group. Methods Thirty-six rats were randomly divided into 3 groups （n= 12）： normal control group, CIA group and ethyl pyruvate group. Then the 6 rats were dissected at the 6th, 9th week respectively. The expression of HMGB1 was analyzed by immunohistochemistry and Pathology-image analysis software in the cytoplasma. The expression of HMGB1 mRNA with real time-polymerse chain reaction （PCR） was evaluate, and the HMGB1 expression of each group were compared with t-test. Results The immunohistochemical results of HMGB1 showed that the expression intensity in the normal control group, CIA model group and ethyl pyruvate group was 2.1＋0.6, 7.3＋l.2, 6.0＋_1.2 respectively at the 6th week; and 2.2±0.7, 12.4±4.5, 5.5±1.0 at the 9th week respectively. The HMGB1 mRNA real time-PCR results had shown that the relative quantification of the normal control group and CIA model group were 1, 2.865, 2.602 respectively at the 6th week and 1.005, 4.694, 1.729 at the 9th week. At those two points, the I-IMGB1expressions of HMGB1 antagonist group were significantly higher than those of the normal controls （P〈0.05）. In addition, there was statistical significant difference（P〈0.05） in the HMGB1 expression when compared with the placebo group. Furthermore, when the degree of HMGB1 expression among the three groups was compared, the HMGB1 antagonist group was decreased significantly （P〈0.05）. Conclusion The results has demonstrated that HMGB1 could induce inflammation in the synovial tissue of CIA rats, and has provided the rationale that HMBG1 could be the target of treating rheumatoid arthritis （RA）. The results of this study have shown that ethyl pyruvate could antagonize the effect of HMGB1. This finding may provide a new therapeutic target for the treatment of RA.
出处
《中华风湿病学杂志》
CAS
CSCD
北大核心
2012年第8期518-522,F0003,共6页
Chinese Journal of Rheumatology
关键词
关节炎
实验性
大鼠
高迁移率族蛋白质类
滑膜
免疫组织化学
聚合酶链反应
Arthritis, experimental
Rats
High mobility group proteins
Synovial membrane
Immunohistochemistry
Polymerase chain reaction
