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目的观察基质金属蛋白酶(MMP)-9及其组织型金属蛋白酶抑制剂(TIMP)-1、一氧化氮(NO)和诱导型一氧化氮合酶(iNOS)在内毒素诱导性葡萄膜炎(EIU)眼组织中的变化。方法90只Sprague-Dawley(SD)大鼠随机分为实验组(81只)和对照组(9只)。实验组大鼠双后足垫注射伤寒杆菌内毒素(LPS)200μl建立EIU模型;对照组大鼠不予注射。在LPS注射后0、6、12、18、24、48、72、96h和7d分别处死9只实验组大鼠,观察眼部改变并进行组织病理学检查。检测血浆、房水和葡萄膜组织的NO含量和房水蛋白浓度。免疫组织化学方法和计算机图像分析系统检测MMP-9、TIMP-1和iNOS的表达及其平均吸光度[A,旧称光密度(OD)]值。结果虹膜、睫状体的上皮细胞和渗出的炎性细胞表达iNOS、MMP-9和TIMP-1。房水蛋白浓度,血浆、房水和葡萄膜组织中NO含量,以及MMP-9的A值与炎症程度呈正相关,iNOS和TIMP-1的A值与炎症程度无明显相关性。iNOS在LPS注射后6h可见表达,12h达高峰,以后逐渐下降。TIMP-1表达出现于24h,72h时达高峰。结论在EIU发生、发展过程中,MMP-9、TIMP-1、NO、iNOS的含量或表达发生变化,提示它们参与EIU的病理过程。
Objective To observe the effects of matrix metalloproteinase (MMP) -9 and tissue inhibitor of metalloproteinase (TIMP) -1, nitric oxide (NO) and inducible nitric oxide synthase (iNOS) on endotoxin-induced uveitis EIU) eye tissue changes. Methods Ninety Sprague-Dawley rats were randomly divided into experimental group (n = 81) and control group (n = 9). In the experimental group, 200 ul of Salmonella typhimurium endotoxin (LPS) was injected into the double hind foot pad to establish EIU model; the control group rats were not injected. Nine rats in experimental group were sacrificed at 0, 6, 12, 18, 24, 48, 72, 96 h and 7 d after LPS injection. Changes of ocular tissues were observed and histopathological examination was performed. Plasma, aqueous humor and uveal tissue NO content and aqueous protein concentration were measured. The expression of MMP-9, TIMP-1 and iNOS and their average absorbance [A, old optical density (OD)] values were detected by immunohistochemistry and computerized image analysis system. Results The iris, ciliary body epithelial cells and exudative inflammatory cells express iNOS, MMP-9 and TIMP-1. Aqueous protein concentration, NO content in plasma, aqueous humor and uveal tissue, as well as the A value of MMP-9 were positively correlated with the degree of inflammation. There was no significant correlation between the A value of iNOS and TIMP-1 and the degree of inflammation. The expression of iNOS was observed at 6h after LPS injection and peaked at 12h, then decreased gradually. TIMP-1 expression appeared in 24h, 72h reached its peak. Conclusions The content or expression of MMP-9, TIMP-1, NO and iNOS in the occurrence and development of EIU changes, suggesting that they participate in the pathological process of EIU.