深Ⅱ度烫伤大鼠创面基质金属蛋白酶-1,2及其抑制剂表达变化机制的探讨

The regulation mechanisms of MMP-1, 2 and TIMP-1,2 on wound healing after partial thickness scald

目的 观察深Ⅱ度烫伤创面愈合过程中基质金属蛋白酶 1,2 (MMP 1,2 )与金属蛋白酶组织抑制因子 1,2 (TIMP 1,2 )的表达规律以及其变化机制。 方法 150只Wistar大鼠 3 0 %深Ⅱ度烫伤后随机分为 5组 :(1)正常对照组 (n =6) ;(2 )单纯烫伤组 (n =3 6) ;(3 )激酶激活剂BDM组 (n =3 6) ;(4)蛋白激酶C制剂H7组 (n =3 6) ;(5)c fos抗体组 (n =3 6)。分别于伤后 3、6h和 1、3、7、14d采取创面皮肤标本 ,检测创面组织c fosmRNA与c fos蛋白的表达 ,以及MMP 1,2 /TIMP 1,2相应变化。 结果 伤后 3～ 6h ,c fosmRNA与c fos蛋白的表达明显增多 ,随后逐渐下降。MMP 1,2 /TIMP 1,2的表达滞后于前者 ,在伤后 3～ 7d表达较高 ,MMP 2 /TIMP 2的表达增加显著。使用BDM后 ,烫伤 3～6h组织c fosmRNA与c fos蛋白的表达增加 ,MMP 1,2 /TIMP 1,2的表达也增强。使用H7后 ,c fosmRNA与c fos蛋白的表达被抑制 ,MMP 1,2 /TIMP 1,2的表达随之减弱。c fos抗体中和内源性c fos蛋白表达后 ,MMP 1/TIMP 1,2的表达受抑制 ,MMP 2的变化不明显。 结论 创面愈合过程中 ,MMP 1,2和TIMP 1,2的表达与蛋白激酶信号途径的激活有密切关系 ,c fos调节MMP 1和TIMP 1和TIMP 2的含量 ,完成对愈合的调控。

Objective To observe the changes of matrix metalloproteinase 1, 2/tissue inhibitor of metalloproteinase 1, 2 (MMP 1, 2 and TIMP 1, 2) in granulationtissue after 30% TBSA deeper partial thickness scald, and explore the regulation mechanism of MMP 2/TIMP 2 during wound healing. Methods 150 male Wistar rats were randomly divided into 5 groups as follows: (1) normal control (n=6); (2) injured control group (n=36): which is subdivided into postburn 3 h, 6 h, 1 d, 3 d, 7 d and 14 d groups, respectively; (3) BDM group (n=36): intravenous injected of 400 mg 2,3 butanedione monoxime in each rat was done after anesthesia;(4) H7 group (n=36): Each rat was intravenous injected of 0 2 mg 1 5 isoquinolinyl sulfony 2 methylpiperazine after anesthesia;(5) anti c fos group (n=36): Each rat was intravenous injected of 5 μg c fos monoclony antibody after anesthesia. The immunohistochemistry staining technique and the reverse transcription polymerase chain reaction (RT PCR) were used for detectting. Results The expression of c fos mRNA and protein was increased from 3 to 6 hours post burn, and then decreased. The expression of MMP 1, 2/TIMP 1, 2 was delayed to 3 days post burn compared with the expression of c fos mRNA and protein. Treatment with BDM induced to raise c fos mRNA and protein expression. The expression of MMP 1, 2/TIMP 1, 2 was also increased accordingly. However, following treatment with H7 inhibited the expression of c fos mRNA and protein, MMP 1, 2/TIMP 1, 2 proteins expression decreased. Exogenous c fos antibody could inhibit endogenous c fos protein expression and the expression of MMP 1/TIMP 1, 2 decreased, but MMP 2 has no notable changes. Conclusions The expression of MMP 1, 2 and TIMP 1, 2 has closely relation protein kinases activated signaling pathways. The expression changes of MMP 1 and TIMP 1/ TIMP 2 depend on c fos expression. Oncogenes play an important role in the change process of wound matrix degradation and remodeling.