米诺环素对大鼠角膜碱烧伤炎症反应调控的机制研究  

Regulation mechanism of minocycline on inflammatory reactions of rats corneal alkali burn

作　　者：张雷[1] 李康[2] 武斌[1] ZHANG Lei;LI Kang;WU Bin(Department of Burn and Plastic Surgery,Affiliated Hospital of Yan'an University,Yan'an Shaanxi 716000,China;Department of Orthopaedics,Yan'an People's Hospital,Yan'an Shaanxi 716000,China)

机构地区：[1]延安大学附属医院烧伤整形手外科,陕西延安716000 [2]陕西省延安市人民医院骨科四病区,陕西延安716000

出　　处：《临床和实验医学杂志》2019年第21期2256-2259,共4页Journal of Clinical and Experimental Medicine

基　　金：陕西省自然科学基础研究计划项目(编号:2014JM4199)

摘　　要：目的探究米诺环素对大鼠角膜碱烧伤炎症反应的调控机制。方法将40只SD大鼠随机分为4组(n=10),分别为正常对照组、模型组、0.1%米诺环素组和0.5%米诺环素组。采用角膜贴敷氢氧化钠(Na OH)滤纸法构建中度角膜(右眼)碱烧伤动物模型,正常对照组以蒸馏水替代;0.1%米诺环素组和0.5%米诺环素组于建模24 h后予以0.1%、0.5%米诺环素滴眼液滴眼,1滴/次,4次/d,连续滴药14 d,对照组和模型组以生理盐水替代。记录各组大鼠角膜混浊程度、炎症指数和中性粒细胞计数,采用HE染色观察角膜组织病理变化,酶联免疫吸附(ELISA)检测大鼠房水中白介素(IL-6、IL-1β)、肿瘤坏死因子α(TNF-α)的含量,Western blot检测角膜组织中血管内皮生长因子(VEGF)、基质金属蛋白酶-2(MMP-2)、核转录因子-κB(NF-κB)、Toll样受体(TLR-2、TLR-4)的表达。结果与对照组相比,模型组角膜混浊程度评分炎性指数、中性粒细胞计数、房水中IL-6、IL-1β及TNF-α水平、角膜组织中VEGF、MMP-2、NF-κB、TLR-2、TLR-4的表达均显著上升(P<0.05)。米诺环素干预后,角膜碱烧伤大鼠水肿基本消失,混浊程度明显减轻,瞳孔隐约可见,新生血管数量明显较少;HE染色显示,角膜上皮愈合,水肿基本消退,胶原纤维排列相对整齐,有极少数新生血管生成;且角膜混浊程度评分、炎性指数、中性粒细胞计数、房水中IL-6、IL-1β及TNF-α水平、角膜组织中VEGF、MMP-2、NF-κB、TLR-2、TLR-4的表达均显著下降(P<0.05),但两种浓度米诺环素组上述改变差异无显著性(P>0.05)。结论米诺环素对大鼠角膜碱烧伤有一定的治疗作用,可能与下调炎症反应、抑制新生血管生成有关,但0.1%米诺环素和0.5%米诺环素的作用无显著差异。Objective To explore regulation mechanism of minocycline on inflammatory reactions of rat corneal alkali burn.Methods 30 SD rats were randomly divided into model group,0.1%minocycline group and 0.5%minocycline group,10 cases in each group.The corneal application NaOH filtering paper method was performed to construct medium(right eye)corneal alkali burn animal models.At 24 after modeling,0.1%minocycline group and 0.5%minocycline group were given eye drops containing 0.1%and and 0.5%minocycline(1 drop/once,4 times/d).And continuous drip lasted for 14 days.The model group was replaced by normal saline.Corneal opacity degree,inflammatory indexes and neutrophil count in each group were recorded.HE staining was performed to observe pathological changes of cornea tissue.ELISA was performed to detect contents of interleukin(IL-6,IL-1β)and tumor necrosis factorα(TNF-α).Western blot was performed to detect expression of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2(MMP-2),nuclear transcription factor-κB(NF-κB)and toll-like receptors(TLR-2,TLR-4).Results Compared with the control group,corneal opacity score inflammatory index,neutrophil count,IL-6,IL-1βand TNF-αlevels in the aqueous humor,VEGF,MMP-2,NF-κB in the corneal tissue,the expression of TLR-2 and TLR-4 in the model group increased significantly(P<0.05).After minocycline intervention,edema of corneal alkali burn rats was basically disappeared,opacity degree was significantly decreased,pupil was faintly visible,and number of new vessels was significantly less.HE staining showed that corneal epithelium was healed,edema was basically subsided,and arrangement of collagen fibers was relatively neat.There was very little neovascularization.Corneal opacity score,inflammatory indexes,neutrophil count,levels of IL-6,IL-1βand TNF-αin aqueous humor,expressions of VEGF,MMP-2,NF-κB,TLR-2 and TLR-4 in corneal tissue were significantly decreased(P<0.05).However,there was no significant difference in the above changes among groups of different concen

关 键 词：大鼠 角膜碱烧伤 米诺环素 炎症反应 新生血管生成 

分 类 号：R779.1[医药卫生—眼科]