INF-γ对体外培养的系膜细胞HMGB1和MMP-2表达的影响

doi:河北省科技厅自然科学基金资助课题（C20070

INF-γ对体外培养的系膜细胞HMGB1和MMP-2表达的影响

刘淑霞，郭惠芳，郝军，张玉军，唐丽娟，陈宁，段惠军

河北医科大学基础医学院病理学教研室，河北石家庄 050017

收稿日期:2008-07-23 修回日期:1900-01-01 出版日期:2009-01-28 发布日期:2009-01-28
通讯作者: 段惠军

Effect of INF-γ on expressions of HMGB1 and MMP-2 protein in mesangial cells

LIU Shu-xia,GUO Hui-fang,HAO Jun,ZHANG Yu-jun,TANG Li-juan,CHEN Ning,DUAN Hui-jun

Department of Pathology,School of Basic Medical Sciences,Hebei Medical University,Shijiazhuang 050017，China

Received:2008-07-23 Revised:1900-01-01 Online:2009-01-28 Published:2009-01-28
Contact: DUAN Hui-jun

摘要/Abstract

摘要： 目的：探讨INF-γ对体外培养的人肾小球系膜细胞高迁移率〖JP3〗族蛋白1（HMGB1）和基质金属蛋白酶2〖JP〗（MMP-2）表达的影响及可能机制，为系统性红斑狼疮（SLE）肾脏损害的治疗提供依据。方法：将常规培养的人肾小球系膜细胞分为正常对照组和INF-γ刺激组，于6、12和24 h收集细胞和上清，RT-PCR和免疫细胞化学法检测HMGB1 mRNA和蛋白表达；双抗夹心ELISA法检测细胞培养上清中HMGB1和MMP-2表达；免疫细胞化学法检测细胞中增殖细胞核抗原（PCNA）和MMP-2表达。结果：与对照组比较，INF-γ刺激组系膜细胞PCNA蛋白表达强度增加，INF-γ刺激12和24 h时系膜细胞中HMGB1 mRNA水平明显增加(P<0.01)，HMGB1蛋白表达强度增加；培养上清中HMGB1浓度明显增加(P<0.01)；与对照组比较，INF-γ刺激组系膜细胞中MMP-2表达强度增加，培养上清中MMP-2蛋白水平升高(P<0.01)；培养上清中HMGB1 浓度与MMP-2蛋白表达量呈正相关关系（r=0.915，P<0.01）。结论：INF-γ通过促进系膜细胞合成和分泌HMGB1而上调MMP-2的表达，可能与SLE肾脏损伤有关联。

关键词: 干扰素Ⅱ型, 高迁移率族蛋白质类, 明胶酶A

Abstract: Abstract:Objective To investigate the effect of INF-γ on the expression of high mobility group box 1 (HMGB1) and MMP-2 protein in mesangial cells(MC) and its possible mechanism in order to provide basis for treatment of renal injury in systemic lupus erythematosus (SLE). Methods Human MC induced by 5 ng•L^-1 INF-γ were collected in 6,12 and 24 h respectively,as well as cells in normal control group in vitro. The expressions of HMGB1mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry.The expressions of proliferation cell nuclear antigen (PCNA) and MMP-2 protein were detected by immunocytochemistry.ELISA was used to determine the levels of HMGB1 and MMP-2 in supernatant.Results Compared with control group,the PCNA protein expression was up-regulated in INF-γ group,the HMGB1 mRNA and protein expressions in MC obviously increased,the HMGB1 content in supernatant increased significantly.Compared with control group,MMP-2 protein in MC and supernatant in INF-γ group increased; there was positive correlation between HMGB1 and MMP-2 protein in supernatant (r=0.915,P<0.01).Conclusion INF-γ can up-regulate the expression of MMP-2 by promoting synthesis of MC and secretion of HMGB1,it might play an important role in renal injury of SLE.

Key words: interferon　type Ⅱ, high mobility group proteins, gelatinase A

中图分类号:

R363

刘淑霞，郭惠芳，郝军，张玉军，唐丽娟，陈宁，段惠军. INF-γ对体外培养的系膜细胞HMGB1和MMP-2表达的影响[J]. J4, 2009, 35(1): 47-50.

LIU Shu-xia,GUO Hui-fang,HAO Jun,ZHANG Yu-jun,TANG Li-juan,CHEN Ning,DUAN Hui-jun. Effect of INF-γ on expressions of HMGB1 and MMP-2 protein in mesangial cells[J]. J4, 2009, 35(1): 47-50.

[1]	潘颖，朱丹，司乃文，张文颖. 卵巢上皮性癌组织RhoC和MMP-2蛋白的表达及其临床意义[J]. J4, 2008, 34(6): 1042-1045.
[2]	马岩，李晓林，陆艳娟,李晓梅，刘坤，高航，张海英，马颖哲，韩智冲，张蒙. 黄蘑碱溶性多糖对荷瘤小鼠MMPs表达的调节及其抗肿瘤作用机制[J]. J4, 2008, 34(4): 605-609.
[3]	杨丹，方艳秋，许淑芬，段秀梅，谭岩. 人白细胞介素24基因的克隆及在大肠杆菌中的高效表达[J]. J4, 2008, 34(2): 266-269.
[4]	杨巍，孙婷，龚平生，李修义，龚守良. 重组质粒pIRESEgr-IFNγ的构建及其在Lewis肺癌细胞中的辐射诱导表达[J]. J4, 2007, 33(5): 790-793.
[5]	罗萍，黄兰，卢雪红，张冬梅. 慢性肾小球肾炎患者血清MMP-2表达及临床意义[J]. J4, 2007, 33(4): 741-743.
[6]	郝冰，孟晓萍，杨东华，王莉，郭昊. 普罗布可对冠心病患者血清中MMP-2的抑制作用[J]. J4, 2007, 33(3): 559-561.
[7]	李蕴潜，赵丽艳，李才. 人胶质瘤细胞的侵袭能力和胶原溶解作用的体外研究[J]. J4, 2007, 33(1): 84-87.
[8]	盛辉，苑春莉，周洪澜，王医术. MMP-2和TIMP-2在肺癌组织中的表达及其与微血管密度的关系[J]. J4, 2006, 32(6): 1059-1062.
[9]	杨巍，孙婷，刘淑春，朴春姬，梁硕，李修义，龚守良. 多次小剂量pEgr-IFNγ-Endostatin基因-放射治疗对小鼠Lewis肺癌的抑瘤作用[J]. J4, 2006, 32(4): 547-549.
[10]	梅红，郭玉芳，郑贤红，王心蕊，张丽红，朱桂彬，王建民，李一雷. 整合素α_Vβ₃在细胞外基质调控人黑色素瘤MMP-2表达和活化中的作用[J]. J4, 2006, 32(2): 182-185.
[11]	邢王月，迟宝荣，李淑梅，景瑕斌，孟晓萍. 聚乳酸-聚乙醇酸-聚乙二醇血管支架置入对兔动脉粥样硬化模型血清基质金属蛋白酶2，9的影响[J]. J4, 2006, 32(2): 257-260.
[12]	张志超,孙亚欣,贾明库. 基质金属蛋白酶含量及表达与肝细胞癌侵袭转移的关系[J]. J4, 2006, 32(1): 116-3.
[13]	郭玉芳，梅红，张丽红，刘巍，郑贤红，李一雷. 人乳腺癌组织活化型MMP-2的分布及意义[J]. J4, 2006, 32(1): 103-3.
[14]	王珂，李然伟，顾莲芝，马忠森. 结核分枝杆菌诱导巨噬细胞产生的条件培养基对γ-干扰素信号转导系统的抑制作用[J]. J4, 2006, 32(1): 31-4.
[15]	张宇靖，谷丽萍，华莹，张首杰. 基质金属蛋白酶-2及其组织抑制剂和微血管密度在子宫内膜癌中的表达[J]. J4, 2006, 32(1): 106-4.