MMP-1及TIMP-1在大鼠肠粘连组织中的表达及其意义

Expression of MMP-1 and TIMP-1 in the tissue of intestinal adhesion and its significance in rat

目的通过大鼠肠粘连动物模型,检测基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶抑制物-1(TIMP-1)在肠粘连组织及正常组织中的表达,研究MMP-1和TIMP-1在肠粘连发病中的作用。方法选取SD雄性大鼠70只随机分为3组:10只作为正常组;观察组30只刮伤其回肠浆膜,创面滴无水酒精,钳夹盲肠系膜动脉制成损伤性肠粘连模型;对照组30只大鼠与观察组一样麻醉、开腹,但不动肠组织,直接关腹。观察组及对照组分别于造模后2、7、14d3个时相点各取10只大鼠,处死并分别取小肠浆膜层浆膜粘连面组织,应用免疫组化和图像分析的方法测定MMP-1和TIMP-1在肠粘连组织及正常肠组织中的蛋白表达。结果 (1)造模术后2d,观察组中MMP-1和TIMP-1的含量均升高(P均<0.01),MMP-1升至峰值。(2)造模术后7d,观察组MMP-1含量降低,而TIMP-1的含量升高至峰值,与术后2d及对照组相比,差异均有统计学意义(P均<0.01)。(3)造模术后14d,观察组MMP-1、TIMP-1的含量均下降,与术后2、7d及对照组相比,差异均有统计学意义(P均<0.01)。(4)对照组各时相点两两比较,无明显差异(P均>0.05)。结论 MMP-1、TIMP-1的活性变化是肠粘连形成的重要调节因素之一。

Objective To determine the expressions of matrix metalloproteinase-1（MMP-1）and tissue inhibitor of metalloproteinase-1（TIMP-1） in the tissues of intestinal adhesion and study the role of MMP-1 and TIMP-1 in the pathogenesis of intestinal adhesion of rat.Methods Seventy SD male rats were randomly assigned to three groups：normal group（n=10）,observation group（n=30） and control group（n=30）.In the observation group,the serous membrane of ileum was injured via scrapping;the dehydrated alcohol was dropped on the surface of wound;cecal mesenteric artery was clamped;so the model of traumatic intestinal adhesion of rat was established.In the control group,the anesthesia and laparotomy were same as observation group,but no intestinal tissue was injured,and no mesenteric artery was clamped.In observation group and control group,10 rats were respectively sacrificed at 2 d,7 d,14 d after modeling,and the tissues of serous membrane in adhesive area were taken.The expressions of MMP-1 and TIMP-1 in the intestinal adhesive tissues and the normal intestinal tissues were determined by immunohistochemistry assay and image analysis.Results（1） The expression levels of both MMP-1 and TIMP-1 elevated 2 days after modeling（all P〈0.01）,and MMP-1 level reached to the peak value.（2）Compared with control group and 2 days after modeling in observation group,the MMP-1 expression reduced,but the TIMP-1 expression elevated to the peak value at 7 days after modeling in observation group（all P〈0.01）.（3） Compared with control group and 2 or 7 days after modeling in observation group,the expression levels of both MMP-1 and the TIMP-1 dropped at 14 days after modeling in observation group（all P〈0.01）.（4） The expression levels of both MMP-1 and TIMP-1 in the control group had no significant difference compared between two phases each other（all P〉0.05）.Conclusions The active change of MMP-1/TIMP-1 is one of the important adjustment factors in the formation of intestinal adhesion.