中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
11期
999-1003
,共5页
视网膜色素上皮%光损伤%基质金属蛋白酶%促红细胞生成素
視網膜色素上皮%光損傷%基質金屬蛋白酶%促紅細胞生成素
시망막색소상피%광손상%기질금속단백매%촉홍세포생성소
Retinal pigment epithilium%Photic injury%Matrix metalloproteinase%Erythropoietin
背景 近年来,随着眼科光学诊疗器械和显微手术应用的增多,各种器械的光源所导致的视网膜损伤等问题倍受关注,研究视网膜光损伤可为临床治疗相关疾病提供参考. 目的 研究小鼠光损伤后视网膜色素上皮(RPE)层基质金属蛋白酶2(MMP-2)、MMP-9表达的变化,探讨促红细胞生成素(EPO)对小鼠视网膜光损伤的治疗作用及其相关机制.方法 将52只SPF级BALB/c小鼠采用抽签法随机分为正常对照组4只、单纯光照组24只和EPO预处理组24只,后两组小鼠在自制光照箱内用6000 lx弥散白光照射4h以建立光损伤动物模型.单纯光照组接受6000 lx白光照射4h,EPO预处理组小鼠腹腔注射rhEPO 5000 U/kg(商品单位)后,接受6000 lx白光照射4h.光照后6、12、36、72、96 h和7d采用免疫组织化学法检测各组小鼠RPE中MMP-2和MMP-9的表达.结果 单纯光照组小鼠视网膜组织病理学改变出现早且明显,光照后RPE细胞出现不同程度的水肿和结构紊乱,且随着光照后时间的延长,形态异常的细胞数增加,但EPO预处理组在光照后7d未发现类似改变.免疫组织化学法检测显示,正常BALB/c小鼠RPE层MMP-2低表达,单纯光照36 h后,RPE层可观察到大量MMP-2阳性表达细胞,但同一时间点EPO预处理组阳性表达量(A值)明显下降.3个组和不同时间点MMP-2的表达量差异均有统计学意义(F分组=3.68,P=0.04;F时间=9.13,P=0.00).MMP-9免疫组织化学法检测显示,正常RPE层中未见MMP-9的阳性表达,单纯光照6h后,MMP-9开始表达,12h时达高峰并维持高表达状态,96 h后表达量开始下降;EPO预处理组MMP-9表达趋势同单纯光照组,但各时间点MMP-9的表达量均较单纯光照组减少.3个组和不同时间点MMP-9的表达差异均有统计学意义(F分组=3.61,P=0.04;F时间=16.91,P=0.00). 结论 MMP-2、MMP-9在视网膜光损伤过程中发挥着重要作用,推测EPO可能通过抑制MMP-2、MMP-9的表达发挥对视网膜的保护作用.
揹景 近年來,隨著眼科光學診療器械和顯微手術應用的增多,各種器械的光源所導緻的視網膜損傷等問題倍受關註,研究視網膜光損傷可為臨床治療相關疾病提供參攷. 目的 研究小鼠光損傷後視網膜色素上皮(RPE)層基質金屬蛋白酶2(MMP-2)、MMP-9錶達的變化,探討促紅細胞生成素(EPO)對小鼠視網膜光損傷的治療作用及其相關機製.方法 將52隻SPF級BALB/c小鼠採用抽籤法隨機分為正常對照組4隻、單純光照組24隻和EPO預處理組24隻,後兩組小鼠在自製光照箱內用6000 lx瀰散白光照射4h以建立光損傷動物模型.單純光照組接受6000 lx白光照射4h,EPO預處理組小鼠腹腔註射rhEPO 5000 U/kg(商品單位)後,接受6000 lx白光照射4h.光照後6、12、36、72、96 h和7d採用免疫組織化學法檢測各組小鼠RPE中MMP-2和MMP-9的錶達.結果 單純光照組小鼠視網膜組織病理學改變齣現早且明顯,光照後RPE細胞齣現不同程度的水腫和結構紊亂,且隨著光照後時間的延長,形態異常的細胞數增加,但EPO預處理組在光照後7d未髮現類似改變.免疫組織化學法檢測顯示,正常BALB/c小鼠RPE層MMP-2低錶達,單純光照36 h後,RPE層可觀察到大量MMP-2暘性錶達細胞,但同一時間點EPO預處理組暘性錶達量(A值)明顯下降.3箇組和不同時間點MMP-2的錶達量差異均有統計學意義(F分組=3.68,P=0.04;F時間=9.13,P=0.00).MMP-9免疫組織化學法檢測顯示,正常RPE層中未見MMP-9的暘性錶達,單純光照6h後,MMP-9開始錶達,12h時達高峰併維持高錶達狀態,96 h後錶達量開始下降;EPO預處理組MMP-9錶達趨勢同單純光照組,但各時間點MMP-9的錶達量均較單純光照組減少.3箇組和不同時間點MMP-9的錶達差異均有統計學意義(F分組=3.61,P=0.04;F時間=16.91,P=0.00). 結論 MMP-2、MMP-9在視網膜光損傷過程中髮揮著重要作用,推測EPO可能通過抑製MMP-2、MMP-9的錶達髮揮對視網膜的保護作用.
배경 근년래,수착안과광학진료기계화현미수술응용적증다,각충기계적광원소도치적시망막손상등문제배수관주,연구시망막광손상가위림상치료상관질병제공삼고. 목적 연구소서광손상후시망막색소상피(RPE)층기질금속단백매2(MMP-2)、MMP-9표체적변화,탐토촉홍세포생성소(EPO)대소서시망막광손상적치료작용급기상관궤제.방법 장52지SPF급BALB/c소서채용추첨법수궤분위정상대조조4지、단순광조조24지화EPO예처리조24지,후량조소서재자제광조상내용6000 lx미산백광조사4h이건립광손상동물모형.단순광조조접수6000 lx백광조사4h,EPO예처리조소서복강주사rhEPO 5000 U/kg(상품단위)후,접수6000 lx백광조사4h.광조후6、12、36、72、96 h화7d채용면역조직화학법검측각조소서RPE중MMP-2화MMP-9적표체.결과 단순광조조소서시망막조직병이학개변출현조차명현,광조후RPE세포출현불동정도적수종화결구문란,차수착광조후시간적연장,형태이상적세포수증가,단EPO예처리조재광조후7d미발현유사개변.면역조직화학법검측현시,정상BALB/c소서RPE층MMP-2저표체,단순광조36 h후,RPE층가관찰도대량MMP-2양성표체세포,단동일시간점EPO예처리조양성표체량(A치)명현하강.3개조화불동시간점MMP-2적표체량차이균유통계학의의(F분조=3.68,P=0.04;F시간=9.13,P=0.00).MMP-9면역조직화학법검측현시,정상RPE층중미견MMP-9적양성표체,단순광조6h후,MMP-9개시표체,12h시체고봉병유지고표체상태,96 h후표체량개시하강;EPO예처리조MMP-9표체추세동단순광조조,단각시간점MMP-9적표체량균교단순광조조감소.3개조화불동시간점MMP-9적표체차이균유통계학의의(F분조=3.61,P=0.04;F시간=16.91,P=0.00). 결론 MMP-2、MMP-9재시망막광손상과정중발휘착중요작용,추측EPO가능통과억제MMP-2、MMP-9적표체발휘대시망막적보호작용.
Background Increase in ophthalmic optical medical instruments and microsurgical applications leads to retinal photochemical damage and other problems delivery of a variety of devices,so the in-depth study and understanding of its pathogenesis after retina light damage can provide a reference for the clinical treatment of related diseases.Objective This study was to investigate the therapeutic effect and relative mechanism of erythropoietin (EPO) on mouse retina photic injury by studying the expression of matrix metalloproteinases-2 (MMP-2)and MMP-9.Methods Fifty-two SPF BALB/c mice were randomized into normal control group,simple light-induced group and EPO pretreatment group by balloting method.The mice of simple light-induced group and EPO pretreatment group were continuously irradiated with 6000 lx diffuse light for 4 hours in a home-made box to establish the models of light-induced damage;while recombinant human EPO (rhEPO)of 5000 U/kg was intraperitoneally injected prior to the light exposure in the EPO pretreatment group.The expressions of MMP-2 and MMP-9 were examined at 6,12,36,72,96 hours and 7 days following light-exposure by immunohistochemistry.Results Edema and structural disorder of RPE cells appeared inthe simple light-induced group after light-exposure and aggravated with lapse of light-exposure time,but no similar change was seen until 7 days in the EPO pretreatment group.The immunohistochemistry findings showed that the expression of MMP-2(A value)in RPE cells was less in the normal mice.However,a large quantity of positive cells appeared in RPE layer 36 hours after light-exposure.Compared with the simple light-induced group,the positive expression of MMP-2 protein in EPO pretreatment group was significantly decreased,showing statistically significant differences among these three groups and different time points (Fgroup =3.68,P =0 04; Ftime =9.13,P=0.00).There was hardly any MMP-9 expression in the retina of the normal mice.In simple light-induced group,a few of positive cells appeared in RPE layer 6 hours after light-exposure and reached its peak 12 hours following light-exposure.The gradually down-regulation of MMP-9 expression happened 96 hours later following light-irradiation.The expression tendency of MMP-9 in EPO pretreatment group was similar to the simple light-induced group.Significant differences in expressions of MMP-9 were found among different groups and time points (Fgroup=3.61,P =0.04;Ftime =16.91,P=0.00).Conclusions MMP-2 and MMP-9 may be involved in the mechanism of retina photic injury by down-regulating the expression of MMP-2 and MMP-9 in RPE cells.
