MRL/lpr狼疮小鼠肾脏明胶酶表达随增龄变化及意义  

Aging changes of gelatinase activity in kidney tissue of autoimmune MRL/lpr mice

作　　者：蔡广研[1] 陈香美[1] 王兆霞[1] 田月[1] 师锁柱[1] 

机构地区：[1]解放军总医院肾科解放军肾病中心暨重点实验室,北京100853

出　　处：《中华病理学杂志》2002年第5期432-435,共4页Chinese Journal of Pathology

基　　金：国家自然科学基金重点项目资助 ( 39930 2 30 );国家重点基础研究发展规划项目资助 (G2 0 0 0 0 5 70 0 3)

摘　　要：目的　观察不同周龄MRL/lpr狼疮小鼠肾脏明胶酶的表达及其随增龄而发生的活性变化及意义。方法　取 8、16与 2 4周龄MRL/lpr狼疮小鼠的肾组织进行常规病理检测 ,利用含有放射自显影的成像乳胶对冷冻肾组织切片进行原位明胶酶活性的检测 ;利用免疫组织化学检测肾脏明胶酶A与明胶酶B的蛋白表达变化 ,十二氨基磺酸钠 聚丙烯酰胺凝胶电泳 (SDS PAGE)明胶酶谱法检测肾脏明胶酶A、B的活性变化。结果　 8周龄狼疮小鼠肾组织中仅在血管处检测到明胶酶的活性 ;16与2 4周龄狼疮小鼠肾小球内明胶酶的净活性明显增加 ,在肾小球以及肾小管间质上也可检测到明胶酶的活性。乙二胺四乙酸 (EDTA)能够抑制肾脏明胶酶的活性。免疫组织化学与SDS PAGE明胶酶谱法结果显示其肾组织中明胶酶A与明胶酶B的蛋白质表达及活性均明显增加。结论　明胶酶A、B在自发性狼疮小鼠肾炎中的表达及活化随增龄均明显增加 。Objective To determine whether increased expressions of gelatinases occur with aging in vivo in kidney tissue of autoimmune MRL/lpr mice by in situ zymography. Methods MRL/lpr mice at the age of 8 weeks, 16 weeks and 24 weeks were investigated. Kidney protein extracts were compared for activities of MMP-2/9 by gelatin zymography. Immunohistochemistry and SDS-PAGE gelatin zymography were used to determine the expressions and activities of gelatinase A(MMP-2) and gelatinase B(MMP-9). To determine the net gelatinase activities in murine lupus kidney, in situ zymography was used with autoradiographic emulsion as substrate. Results Both gelatinase A and B were seldom detected in the kidney tissue in 8 week old mice, Increased expressions of both latent and activated form enzymes of MMP-2/9 were identified in kidney extraction by SDS-PAGE gelatin zymography and immunohistochemical staining showed both MMP-2 and MMP-9 were obviously up-regulated within glomerulus as well as tubular-interstitium in mice at the age of 16 and 24 weeks. In situ zymography showed markedly increased gelatinase activities in kidney tissue consistent with the results of immunohistochemical staining, it is mainly derived from MMPs and inhibited by EDTA but not by PMSF or aprotinin. Conclusions These in vivo results suggested that MMP-2/-9 expressions were significantly up-regulated with aging in murine lupus nephritis, which may play an important role in promoting the remodeling formation of ECM and thus contribute to the progression of renal damage in this model.

关 键 词：狼疮肾炎 明胶酶 原位酶谱法 动物实验 

分 类 号：R593.242[医药卫生—内科学]