多西环素对角膜瘢痕形成的影响  

Effect of doxycycline on corneal scar formation

作　　者：周佳欣 邹文进[1] Zhou Jiaxin;Zou Wenjin(Department of Ophthalmology,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院眼科,南宁530021

出　　处：《广西医科大学学报》2022年第4期606-611,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81160119);广西重点研发计划资助项目(No.AB20238003)。

摘　　要：目的:研究多西环素对角膜瘢痕形成的影响。方法:(1)体外实验:收集人眼角膜基质分离并培养,使用免疫荧光双染鉴定所提取到的细胞为人角膜细胞。多西环素组在传代培养后加入重组人转化生长因子-β1(TGF-β1)诱导人角膜细胞分化成角膜肌成纤维细胞,培养2 h后加入浓度为20 ng/mL的多西环素溶液,另设不加入任何药物的对照组和只加入TGF-β1的模型组,放入培养箱继续培养48 h。蛋白免疫印迹(Western blotting)检测CollagenⅠ和Fibronectin蛋白表达;实时荧光定量PCR(RT-qPCR)法检测CollagenⅠ和Fibronectin mRNA相对表达量。(2)体内实验:采用36只雌性SD大鼠构建角膜碱烧伤模型,随机分为对照组和多西环素组,每组18只。采用体视显微镜观察大鼠角膜恢复情况并在第3、第7、第14天进行角膜混浊度评分,角膜取材后做HE染色观察大鼠角膜组织结构变化。结果:模型组细胞Fibronectin、CollagenⅠmRNA和蛋白相对表达量均较对照组升高(均P<0.05),多西环素组细胞Fibronectin、CollagenⅠmRNA和蛋白相对表达量均较模型组降低(均P<0.05)。与对照组相比,大鼠角膜碱烧伤后第7、第14天,多西环素组大鼠角膜混浊度评分更低(P<0.05);HE染色图片,各个时间点多西环素组大鼠角膜基质结构更规整,第7天多西环素组上皮修复更佳、炎症细胞浸润更少。结论:多西环素可以抑制人角膜细胞分化为肌成纤维细胞,促进大鼠角膜碱烧伤后角膜愈合,减轻角膜瘢痕。Objective:To study the effect of doxycycline on the formation of corneal scar.Methods:(1)In vitro experiments:Human corneal stroma was collected,isolated and cultured,and the extracted cells were identified as human corneal cells using immunofluorescence double staining.In the doxycycline group,recombinant human transforming growth factor-β1(TGF-β1)was added to induce differentiation of human corneal cells into corneal myofibroblasts after culturing for 2 h,and doxycycline solution at a concentration of 20 ng/mL was added after 2 h.A control group without any drug and a model group with only TGF-β1 added were set up and incubated for 48 h.Western blotting was used to detect the expression levels of Collagen I and Fibronectin protein.Real-time quantitative PCR(RT-qPCR)was used to detect the relative expression levels of CollagenⅠand Fibronectin mRNA.(2)In vivo experiments:36 female SD rats were used to construct corneal alkali burn models and randomly divided into control and doxycycline groups,with 18 rats in each group.The recovery of the corneal was observed using a stereomicroscope,and corneal cloudiness was scored on day 3,day 7 and day 14.Results:The relative expressions of Fibronectin,Collagen mRNA and protein were higher in the model group compared with the control group(P<0.05),and the relative expression of Fibronectin,Collagen mRNA and protein were lower in the doxycycline group compared with the model group(P<0.05).Compared with the control group,the corneal cloudiness scores of rats in the doxycycline group were lower on days 7 and 14 after corneal alkali burns(P<0.05).HE staining pictures showed that the corneal stromal structure was more regular in the doxycycline group at all time points.On day 7,the epithelial repair was better in the doxycycline group,and the inflammatory cell infiltration was also less.Conclusion:Doxycycline can inhibit the differentiation of human corneal cells into myofibroblasts,promote corneal healing and reduce scar formation after corneal alkali burns in rats.

关 键 词：角膜瘢痕 角膜碱烧伤 多西环素 转化生长因子Β1 角膜肌成纤维细胞 

分 类 号：R779.1[医药卫生—眼科]