摘要
背景:细胞因子是一类多功能强效的细胞生长调节因子,调节多种细胞的生长过程。研究表明细胞因子作为炎症介质在眼部炎症反应中占据重要地位,但其在外伤性白内障术后的炎症反应中的动态变化目前研究较少。目的:探讨兔外伤性白内障囊外摘除术后房水中白细胞介素1、白细胞介素6及肿瘤坏死因子α表达水平的动态变化及其与眼内炎症反应的关系。设计:随机对照的动物实验。单位:暨南大学医学院附属深圳眼科中心深圳市眼科医院。材料:实验于2004-03/2006-03在暨南大学医学院附属深圳眼科中心研究所完成。选择健康新西兰成年家兔15只,30只眼。普通级,体质量2.5～3.0kg,雌雄不限(暨南大学第二临床医学院动物中心提供,许可证号:SYXK(粤)2005-0061)。实验前检查家兔双眼未见异常。将家兔随机分成3组,分别为正常对照组、外伤对照组及外伤手术组(行囊外晶体摘除术),每组5只,10只眼。白细胞介素1、白细胞介素6及肿瘤坏死因子α的检测试剂盒由深圳市雅为生物科技有限公司提供。方法:将外伤对照组和外伤手术组家兔全麻(静脉注射10%乌拉坦,1mL/kg)后,用5号针头自角膜缘穿刺进入前房,划破晶体前囊膜约5mm,制成双眼外伤性白内障动物模型。正常对照组家兔正常喂养。外伤对照组及外伤手术组家兔造模成功后双眼均滴用普通抗生素眼药水清洁结膜囊,3次/d。造模成功后第3天全麻(静脉注射10%乌拉坦,1mL/kg)外伤手术组的家兔,于手术显微镜下由制作动物模型的同一手术者施行常规双眼白内障囊外摘除术,进入前房后行水平截囊或利用外伤性前囊膜破口,进行水分离,娩出晶体核,冲洗干净晶体皮质,缝合切口。于手术当天及术后1d,3d,7d,14d分别检查外伤对照组和外伤手术组家兔前房炎症反应情况并分别抽取3组家兔的房水0.2mL做房水细胞计数及分类,同时通过双抗体ELASA法测定3组家兔的房水中细胞因子白细胞介素1,白细胞介素6和肿瘤坏死因子α的表达水平和动态变化。主要观察指标:3组家兔术后不同时间房水中白细胞总数及白细胞介素1、白细胞介素6和肿瘤坏死因子α含量比较。结果:①正常对照组家兔在术后1d,3d,7d,14d房水中白细胞总数分别为(2.4±0.7)×106/L,(2.2±0.5)×106/L,(2.8±0.8)×106/L和(2.0±0.5)×106/L;外伤对照组分别为(19.7±7.3)×106/L,(28.1±9.6)×106/L,(14.2±5.6)×106/L和(8.4±3.8)×106/L。外伤手术组分别为(65.3±14.5)×106/L,(79.8±12.7)×106/L,(21.7±8.2)×106/L和(12.4±4.1)×106/L。外伤对照组和外伤手术组明显高于正常对照组(F=22.5,27.9,11.6,8.4,P<0.05)。②术后1～14d外伤对照组和外伤手术组家兔房水中的白细胞介素1、白细胞介素6及肿瘤坏死因子α含量均明显高于正常对照组(P<0.05),外伤手术组与外伤对照组之间差异也具有显著性(P<0.05),手术当天3组家兔之间比较差异无显著性(P>0.05)。③各细胞因子的含量在术后7d时达到高峰,而后逐渐降低,至14d时明显降低,外伤对照组和外伤手术组仍明显高于正常对照组(P<0.05)。结论:外伤及手术后眼内炎症反应的严重程度与房水中细胞因子白细胞介素1、白细胞介素6及肿瘤坏死因子α表达水平的动态变化密切相关,细胞因子是外伤性白内障术后眼内炎症反应的重要介质。
BACKGROUND ： Cytokine is a poly-functional and effective regulator factor to regulate growth of multiple cells. Research- es suggest that, as an inflammatory medium, cytokines play a key role in inflammatory reaction of eye; however, there are rare studies on dynamic changes after traumatic cataract.
OBJECTIVE： To investigate the relationship among inflammatory reaction and dynamic changes of interleukin 1 （IL-1）, IL-6 and tumor necrosis factor alpha （TNF-α） in aqueous humor of rabbits after extracapsular cataract extraction.
DESTGN： Randomized controlled animal study
SETTING ： Shenzhen Municipal Ophthalmology Hospital, Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University.
MATERIALS ： The experiment was carried out in Institute of Shenzhen Municipal Ophthalmology Center Affiliated to Medical College of Jinan University from March 2004 to March 2006. Fifteen healthy adult New Zealand rabbits of 30 eyes, general grade, weighting 2.5-3.0 kg, of either gender, were provided by Animal Center of the Second Clinical Medical College of Jinan University [certification： SYXK （yue） 2005-006]. EyeS of rabbits were normal before experiment. All rabbits were randomly divided into normal control group, traumatic control group and operative group with 5 in each group and in total of 10 eyes in each group. IL-1, IL-6 and TNF-α kits were provided by Shenzhen Yawel Blotechnology Com; pany Limited.
METHODS ： Rabbits in traumatic control group and operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, 5# needle was punctured from corneal Ilmbus to anterior chamber to scarify anterior membrane of lens about 5 mm to establish animal models of traumatic cataract of oculus uterque. Rabbits in normal control group were fed normally. After successful modeling, common antibiotic eyedrops was used to clean conjunctival sac of rabbits in traumatic control group and operative group 3 times a day. On the 3^rd day of successful modeling, rabbits in operative group were totally anesthetized with intravenous injection of 10% 1 mL/kg urethan, and then, they undertook extracapsular cataract extraction of oculus uterque： horizontally intercepting bladder or watedy separating with breakage of anterior bladder membrane, expulsing nucleus of lens, washing lens cortex and suturing incisions. On the operative day and on the 1^st, 3^rd, 7^th and 14^th days after, operation, inflammatory reaction of antedor chamber in traumatic control group and operative group was measured and 0.2 mL aqueous humor was extracted from rabbits in three groups to count and classify cells; meenwhlle, expressed level and dynamic changes of IL-1, IL-6 and TNF-α in cytokines of aqueous humor were measured with double-antibodies ELASA technique.
MAIN OUTCOME MEASURES ： Total numbors of leucocytes and contents of IL-1, IL-6 and TNF-α in aqueous humor of rab= bits in three groups after operation.
RESULTS： ① On the 1^st, 3^rd, 7^th and 14^th days after operation, numbers of leucocytes were （2.4±0.7）×10^6/L, （2.2±0.5）× 10^6/L, （2.8±0.8）×10^6/L and （2.0±0.5）×10^6/L in aqueous humor; （19.7±7.3）×10^6/L, （28.1±9.6）×10^6/L, （14.2±5.6）×10^6/L and （8.4±3.8）×10^6/L in traumatic control group; （65.3±14.5）×10^6/L, （79.8±12.7）×10^6/L, （21.7±8.2）×10^6/L and （12.4±4.1）×10^6/L in operative group. In addition, numbers of leucocytes were more in traumatic control group and operative group than those in normal control group （F =22.5, 27.9, 11.6, 8.4;P 〈 0.05）. ② Within 1-14 days after operation, contents of IL-1, IL-6 and TNF-α in aqueous humo were higher in traumatic control group and operative group than those in normal control group （P 〈 0.05）, and there was a significant difference between traumatic Control group and operative group （P 〈 0.05）; however, there was no significant difference among three groups on the operative day （P 〉 0.05）, ③ Contents of cytokines reached peak on the 7^th day after operation, decreased gradually, and reached the lowest value on the 14^th day. Contents in traumatic control group and operative group were higher than those in normal control group （P 〈 0.05）.
CONCLUSION： The intraocular inflammation after lens extraction is closely related to the dynamic changes of IL-1, IL-6 and TNF-α levels in aqueous humor. Cytokine may be one of crucially inflammatory agents in the eyes after traumatic cataract.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第2期383-386,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
深圳市科技局基金资助项目(200304194)~~
