人TIMP-1转基因小鼠随增龄肾组织炎症反应变化的意义  

Inflammatory reaction changes with aging in kidneys of human TIMP-1 transgenic mice

作　　者：张雪光[1] 陈香美[1] 洪权[1] 尚希瑶[1] 师锁柱[1] 尹忠[1] 蔡广研[1] 

机构地区：[1]解放军总医院肾科全军肾脏病中心暨重点实验室,北京市100853

出　　处：《中华老年医学杂志》2006年第12期916-921,共6页Chinese Journal of Geriatrics

基　　金：国家自然科学基金创新群体科学基金(30121005);国家自然科学基金(30300161);北京市自然科学基金(7032045)

摘　　要：目的利用人基质金属蛋白酶组织抑制物-1(tissue inhibitor of metalloproteinase-1,TIMP-1)转基因小鼠,研究了IMP-1在肾脏器官衰老过程中的作用。方法对3、12、24月龄人TIMP-1转基因小鼠和野生型小鼠肾组织石蜡切片行PAS染色；免疫荧光法检测肾组织内F4/80阳性细胞数；Western印迹方法检测肾组织内TIMP-1、TIMP-2、基质金属蛋白酶(matrix metalloproteinase,MMP)-9、MMP-2、细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)、转化生长因子β1(transforming growth factorβ1,TGF-β1)、Ⅲ型和Ⅳ型胶原蛋白质表达；明胶酶谱法检测明胶酶活性。反向酶谱法检测TIMP-1的活性。结果两种基因型小鼠随增龄肾组织内出现局灶性纤维化改变。24月龄,与野生型小鼠比较,转基因小鼠肾组织内TIMP-1表达及活性上调[(1．10±0．18)对(0．62±0．09)；(50．75±7．25)对(20．64±3．50),P＜0．05]；明胶酶的表达及活性降低[(MMP-2：(2．08±0．20)对(3．39±0．43)；MMP-9：(4．02±0．82)对(6．72±1．40),P＜0．05]；Ⅲ型和Ⅳ型胶原[(0．72±0．11)对(0．57±0．09)；(0．84±0．13)对(0．6±0．11),P＜0．05]、ICAM-1和TGF-β1的表达上调[(0．72±0．12)对(0．53±0．07)；(0．69±0．12)对(0．45±0．09),P＜0．05],F4/ 80阳性细胞增多[(18．8±4．4)对(12．7±3．6),P＜0．05]。结论TIMP-1通过上调ICAM-1、增强炎症,促进衰老相关肾脏纤维化。Objective To explore the role of tissue inhibitor of metalloproteinase-1 （TIMP-1） during renal senescence by using human TIMP-1 transgenic mice. Methods Renal histological changes of wild type mice and transgenic mice at the age of 3, 12, 24 months were observed by periodic acid-schiff （PAS） staining of paraffin sections. The numbers of F4/80 positive cells were detected by immunofluorescence. The protein expressions of TIMP-1, TIMP-2, matrix metalloproteinase （MMP） -9, MMP-2, intercellular adhesion molecule-1 （ICAM-1）, transforming growth factor β1 （TGF-β1）, collagen Ⅲ and collagen Ⅳ were detected by Western blot. The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively. Results Focal renal fibrosis was found in two genotypes with aging. At the age of 24 months, compared with wild type, in kidneys of transgenic type, the expressions and activities of gelatinases were dowregulated （MMP-2： 2.08±0. 20 vs. 3.39±0. 43; MMP-9： 4.02±0. 82 vs. 6.72±1.40, all P〈0.05）; the expressions of collagen Ⅲ, collagen Ⅳ, ICAM-1, and TGF-β1 were upragulated （0. 72±0.11 vs. 0. 57±0. 09; 0. 84±0. 13 vs. 0. 6±0. 11; 0. 72±0.12 vs. 0.53±0. 07; 0. 69±0.12 vs. 0.45±0.09, all P〈0. 05）, and the numbers of F4/80 positive cells were increased （18.8±4.4 vs. 12.7±3.6, P〈0. 05） with the upregulated expression and activity of TIMP-I（1.10±0.18vs. 0.62±0.09; 50.75±7.25 vs. 20.64±3.50, P〈0.05）. Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.

关 键 词：金属蛋白酶类组织剂 肾 纤维化 炎症 

分 类 号：R692[医药卫生—泌尿科学]