中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2009年
3期
260-265
,共6页
徐韶琳%高志卓%王英%陈杰
徐韶琳%高誌卓%王英%陳傑
서소림%고지탁%왕영%진걸
青光眼%巩膜%明胶酶类%金属蛋白酶类组织抑制剂
青光眼%鞏膜%明膠酶類%金屬蛋白酶類組織抑製劑
청광안%공막%명효매류%금속단백매류조직억제제
Glaucoma%Sclera%Gelatinases%Tissue inhibitor of metalloproteinases
目的 探讨大鼠慢性高眼压模型筛板区巩膜组织中基质金属蛋白酶(MMP)及其抑制剂(TIMP)的表达与青光眼发病机制的关系.方法 实验研究.将43只Wistar大鼠的左眼成功地建立为慢性高眼压模型,对侧右眼为对照组.采用计算机随机数字表法将实验大鼠分为3组,分别应用免疫组织化学法(13只鼠)、免疫印迹法(15只鼠)及逆转录聚合酶链反应(RT-PCR)法(15只鼠)检测大鼠筛板区巩膜组织中MMP-2、MMP-9、TIMP-1、TIMP-2的表达情况.应用SAS统计学软件,对免疫印迹法和RT-PCR法榆测MMP-2、MMP-9、TIMP-1、TIMP-2表达的4个定量指标,分别采用具有一个重复测量的两因素设计定量资料的方差分析,以P＜0.01作为差异有统计学意义.结果 (1)免疫组织化学检测结果:实验眼筛板区巩膜组织中细胞核和细胞浆巾MMP-2、MMP-9表达明显,对照眼均未见MMP-2、MMP-9表达;实验眼和对照眼筛板区巩膜组织中细胞核和细胞浆中均未见TIMP-1、TIMP-2表达.(2)免疫印迹法检测结果:平均灰度值:实验眼的MMP-2为193.88±8.84,MMP-9为202.65±8.37,TIMP-1为283.63±5.65,TIMP-2为284.75±5.50;对照眼的MMP-2为117.38±10.76,MMP-9为134.13±5.06,TIMP-1为186.88±7.14,TIMP-2为183.0±5.58.(3)RT-PCR法检测结果:平均灰度值:实验眼的MMP-2为200.50±3.25,MMP-9为200.13±2.95,TIMP-1为201.88±3.14,TIMP-2为195.50±3.55;对照眼的MMP-2为181.88±9.36,MMP-9为181.75±5.85,TIMP-1为179.25±9.21,TIMP-2为179.75±7.12.实验眼与对照眼的表达差异有统计学意义(MMP-2:F=405.55,MMP-9:F=436.11,TIMP-1:F=1167.77,TIMP-2:F=4629.64;均P＜0.01).结论大鼠慢性高眼压模型筛板区巩膜组织中MMP-2、MMP-9、TIMP-1、TIMP-2的表达明显增加.(中华眼科杂志,2009,45:260-265)
目的 探討大鼠慢性高眼壓模型篩闆區鞏膜組織中基質金屬蛋白酶(MMP)及其抑製劑(TIMP)的錶達與青光眼髮病機製的關繫.方法 實驗研究.將43隻Wistar大鼠的左眼成功地建立為慢性高眼壓模型,對側右眼為對照組.採用計算機隨機數字錶法將實驗大鼠分為3組,分彆應用免疫組織化學法(13隻鼠)、免疫印跡法(15隻鼠)及逆轉錄聚閤酶鏈反應(RT-PCR)法(15隻鼠)檢測大鼠篩闆區鞏膜組織中MMP-2、MMP-9、TIMP-1、TIMP-2的錶達情況.應用SAS統計學軟件,對免疫印跡法和RT-PCR法榆測MMP-2、MMP-9、TIMP-1、TIMP-2錶達的4箇定量指標,分彆採用具有一箇重複測量的兩因素設計定量資料的方差分析,以P＜0.01作為差異有統計學意義.結果 (1)免疫組織化學檢測結果:實驗眼篩闆區鞏膜組織中細胞覈和細胞漿巾MMP-2、MMP-9錶達明顯,對照眼均未見MMP-2、MMP-9錶達;實驗眼和對照眼篩闆區鞏膜組織中細胞覈和細胞漿中均未見TIMP-1、TIMP-2錶達.(2)免疫印跡法檢測結果:平均灰度值:實驗眼的MMP-2為193.88±8.84,MMP-9為202.65±8.37,TIMP-1為283.63±5.65,TIMP-2為284.75±5.50;對照眼的MMP-2為117.38±10.76,MMP-9為134.13±5.06,TIMP-1為186.88±7.14,TIMP-2為183.0±5.58.(3)RT-PCR法檢測結果:平均灰度值:實驗眼的MMP-2為200.50±3.25,MMP-9為200.13±2.95,TIMP-1為201.88±3.14,TIMP-2為195.50±3.55;對照眼的MMP-2為181.88±9.36,MMP-9為181.75±5.85,TIMP-1為179.25±9.21,TIMP-2為179.75±7.12.實驗眼與對照眼的錶達差異有統計學意義(MMP-2:F=405.55,MMP-9:F=436.11,TIMP-1:F=1167.77,TIMP-2:F=4629.64;均P＜0.01).結論大鼠慢性高眼壓模型篩闆區鞏膜組織中MMP-2、MMP-9、TIMP-1、TIMP-2的錶達明顯增加.(中華眼科雜誌,2009,45:260-265)
목적 탐토대서만성고안압모형사판구공막조직중기질금속단백매(MMP)급기억제제(TIMP)적표체여청광안발병궤제적관계.방법 실험연구.장43지Wistar대서적좌안성공지건립위만성고안압모형,대측우안위대조조.채용계산궤수궤수자표법장실험대서분위3조,분별응용면역조직화학법(13지서)、면역인적법(15지서)급역전록취합매련반응(RT-PCR)법(15지서)검측대서사판구공막조직중MMP-2、MMP-9、TIMP-1、TIMP-2적표체정황.응용SAS통계학연건,대면역인적법화RT-PCR법유측MMP-2、MMP-9、TIMP-1、TIMP-2표체적4개정량지표,분별채용구유일개중복측량적량인소설계정량자료적방차분석,이P＜0.01작위차이유통계학의의.결과 (1)면역조직화학검측결과:실험안사판구공막조직중세포핵화세포장건MMP-2、MMP-9표체명현,대조안균미견MMP-2、MMP-9표체;실험안화대조안사판구공막조직중세포핵화세포장중균미견TIMP-1、TIMP-2표체.(2)면역인적법검측결과:평균회도치:실험안적MMP-2위193.88±8.84,MMP-9위202.65±8.37,TIMP-1위283.63±5.65,TIMP-2위284.75±5.50;대조안적MMP-2위117.38±10.76,MMP-9위134.13±5.06,TIMP-1위186.88±7.14,TIMP-2위183.0±5.58.(3)RT-PCR법검측결과:평균회도치:실험안적MMP-2위200.50±3.25,MMP-9위200.13±2.95,TIMP-1위201.88±3.14,TIMP-2위195.50±3.55;대조안적MMP-2위181.88±9.36,MMP-9위181.75±5.85,TIMP-1위179.25±9.21,TIMP-2위179.75±7.12.실험안여대조안적표체차이유통계학의의(MMP-2:F=405.55,MMP-9:F=436.11,TIMP-1:F=1167.77,TIMP-2:F=4629.64;균P＜0.01).결론대서만성고안압모형사판구공막조직중MMP-2、MMP-9、TIMP-1、TIMP-2적표체명현증가.(중화안과잡지,2009,45:260-265)
Objective To investigate the expression of matrix metalloproteinases(MMPs)and tissue inhibitors of metallopmteinases(TIMPs)in the sclera of lamina cribrosa in rat chronic elevated intraocular pressure(IOP)and it's the relationship with the pathogenesis of glaucoma.Methods It was an experimental study.Chronic ocular hypertension(OHT)model was induced in the left eyes of 43 Wistar rats by cauterizing episcleral venous while the right eyes were used as control.The rats were divided into three groups:13 for immunohistology,15 for Western blotting,and 15 for RT-PCR by randomly computerized assignment and sacrificed at week 12 after OHT induction.The expression of MMP-2,MMP-9,TIMP-1,and TIMP-2 in the sclera lamina cribrosa was detected using immunohistology,Western blotting,and RT-PCR technique,respectively.Repeated measures ANOVA of two-factor Studies in SAS statistic software were used to analyze the data.Results After 12 weeks,compared with control eyes,the lOP of rat OHT eyes was significantly(F=1519.67,P＜0.01)elevated.The expression of MMP-2 and MMP-9 was increased in cell nucleus and cytoplasm in sclera lamina cribrosa in rat OHT when compared to control eyes with negative expression.No expressions of TIMP-1、TIMP-2 in sclera lamina cribrosa were found in both OHT and control eyes.The mean gray-scale values of MMP-2,MMP-9,TIMP-1,and TIMP-2 in OHT were significantly(F=405.55,F:436.11,F=1167.77,and F=4629.64,P＜0.01)higber than control eye by Western blotting(193.88±8.84 vs 117.38±10.76,202.65±8.37 vs 134,13±5.06.283.63±5.65 vs 186.88±7.14,and 284.75±5.50 vs 183.0±5.58),while by RT-PCR(200.50±3.25 vs 181.88±9.36,200.13±2.95 vs 181.75±5.85,201.88±3.14 vs 179.25±9.21,and 195.50±3.55 vs 179.75±7.12).respectively.Conclusion The increased expression of MMP-2,MMP-9,TIMP-1,and TIMP-2 in lamina cribrosa in rat OHT eye indicates that MMPs and TIMPs may be involved in the pathogenesis of glaucoma.(Chin J Ophthalmol,2009,45:261-266)
