中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
Chinese Journal of Experimental Ophthalmology
2015年
10期
887-891
,共5页
Toll样受体2%单克隆抗体/用法和用量%穿透角膜移植/病理%植片排斥/病理%植片存活/药物作用%动物模型
Toll樣受體2%單剋隆抗體/用法和用量%穿透角膜移植/病理%植片排斥/病理%植片存活/藥物作用%動物模型
Toll양수체2%단극륭항체/용법화용량%천투각막이식/병리%식편배척/병리%식편존활/약물작용%동물모형
Toll-like receptor 2%Antibodies,monoclonal/administration & dosage%Keratoplasty,penetrating/pathology%Graft rejection/pathology%Graft survival/drug effects%Disease models,animal
背景 Toll样受体2(TLR2)在移植免疫相关疾病中的作用越来越受到关注,阻断TLR2对心脏和肾脏移植物的保护作用已有研究报道,但TLR2是否具有抑制角膜移植排斥反应的作用尚未证实.目的 观察TLR2单克隆抗体对同种异体大鼠角膜移植术后植片存活情况的影响.方法 取24只SPF级雌性Wistar大鼠作为受体,12只SPF级SD大鼠作为供体,体质量均为180～220 g.采用同种异体角膜移植的方法在Wistar大鼠右眼实施穿透角膜移植术,制备大鼠角膜移植模型.采用随机数字表法将模型眼分为单纯模型组和TLR2单克隆抗体组,TLR2单克隆抗体组分别于术后0、2、4、6、8d球结膜下注射TLR2单克隆抗体15 μg/30μl,单纯模型组大鼠同法注射等容量生理盐水.术后每日于裂隙灯显微镜下观察各组大鼠角膜植片水肿程度、透明度和新生血管形成情况,参照Holland等的评分标准对排斥反应指数(RI)进行评分;分别于术后第9天、第15天收集各组3只大鼠角膜组织,另取3只正常Wistar大鼠眼球作为正常对照,行常规组织病理学检查.各组取6只大鼠用于生存分析观察.结果 术后1～4 d,裂隙灯显微镜下2个组角膜均轻度水肿,术后9～14d,单纯模型组大鼠角膜植片布满新生血管,可见植片水肿、混浊,而TLR2单克隆抗体组大鼠植片至术后第15天开始混浊.术后5、9、15d,单纯模型组大鼠RI评分均明显高于TLR2单克隆抗体组,差异均有统计学意义(t=4.183、4.954、13.506,均P＜0.05);TLR2单克隆抗体组角膜植存活时间为15.5d,95％可信区间(CI)为14.9～16.1,单纯模型组角膜植片存活时间为9.5d,95％ CI为8.7～10.3,2个组间差异有统计学意义(Z=12.728,P=0.001).角膜组织病理学检查显示,单纯模型组大鼠角膜基质层水肿明显,角膜组织中可见大量炎性细胞浸润和新生血管管腔形成,而在TLR2单克隆抗体组中仅见少量炎性细胞和新生血管.结论 TLR2单克隆抗体能减轻角膜移植术后的炎症反应,延长角膜植片的存活时间.
揹景 Toll樣受體2(TLR2)在移植免疫相關疾病中的作用越來越受到關註,阻斷TLR2對心髒和腎髒移植物的保護作用已有研究報道,但TLR2是否具有抑製角膜移植排斥反應的作用尚未證實.目的 觀察TLR2單剋隆抗體對同種異體大鼠角膜移植術後植片存活情況的影響.方法 取24隻SPF級雌性Wistar大鼠作為受體,12隻SPF級SD大鼠作為供體,體質量均為180～220 g.採用同種異體角膜移植的方法在Wistar大鼠右眼實施穿透角膜移植術,製備大鼠角膜移植模型.採用隨機數字錶法將模型眼分為單純模型組和TLR2單剋隆抗體組,TLR2單剋隆抗體組分彆于術後0、2、4、6、8d毬結膜下註射TLR2單剋隆抗體15 μg/30μl,單純模型組大鼠同法註射等容量生理鹽水.術後每日于裂隙燈顯微鏡下觀察各組大鼠角膜植片水腫程度、透明度和新生血管形成情況,參照Holland等的評分標準對排斥反應指數(RI)進行評分;分彆于術後第9天、第15天收集各組3隻大鼠角膜組織,另取3隻正常Wistar大鼠眼毬作為正常對照,行常規組織病理學檢查.各組取6隻大鼠用于生存分析觀察.結果 術後1～4 d,裂隙燈顯微鏡下2箇組角膜均輕度水腫,術後9～14d,單純模型組大鼠角膜植片佈滿新生血管,可見植片水腫、混濁,而TLR2單剋隆抗體組大鼠植片至術後第15天開始混濁.術後5、9、15d,單純模型組大鼠RI評分均明顯高于TLR2單剋隆抗體組,差異均有統計學意義(t=4.183、4.954、13.506,均P＜0.05);TLR2單剋隆抗體組角膜植存活時間為15.5d,95％可信區間(CI)為14.9～16.1,單純模型組角膜植片存活時間為9.5d,95％ CI為8.7～10.3,2箇組間差異有統計學意義(Z=12.728,P=0.001).角膜組織病理學檢查顯示,單純模型組大鼠角膜基質層水腫明顯,角膜組織中可見大量炎性細胞浸潤和新生血管管腔形成,而在TLR2單剋隆抗體組中僅見少量炎性細胞和新生血管.結論 TLR2單剋隆抗體能減輕角膜移植術後的炎癥反應,延長角膜植片的存活時間.
배경 Toll양수체2(TLR2)재이식면역상관질병중적작용월래월수도관주,조단TLR2대심장화신장이식물적보호작용이유연구보도,단TLR2시부구유억제각막이식배척반응적작용상미증실.목적 관찰TLR2단극륭항체대동충이체대서각막이식술후식편존활정황적영향.방법 취24지SPF급자성Wistar대서작위수체,12지SPF급SD대서작위공체,체질량균위180～220 g.채용동충이체각막이식적방법재Wistar대서우안실시천투각막이식술,제비대서각막이식모형.채용수궤수자표법장모형안분위단순모형조화TLR2단극륭항체조,TLR2단극륭항체조분별우술후0、2、4、6、8d구결막하주사TLR2단극륭항체15 μg/30μl,단순모형조대서동법주사등용량생리염수.술후매일우렬극등현미경하관찰각조대서각막식편수종정도、투명도화신생혈관형성정황,삼조Holland등적평분표준대배척반응지수(RI)진행평분;분별우술후제9천、제15천수집각조3지대서각막조직,령취3지정상Wistar대서안구작위정상대조,행상규조직병이학검사.각조취6지대서용우생존분석관찰.결과 술후1～4 d,렬극등현미경하2개조각막균경도수종,술후9～14d,단순모형조대서각막식편포만신생혈관,가견식편수종、혼탁,이TLR2단극륭항체조대서식편지술후제15천개시혼탁.술후5、9、15d,단순모형조대서RI평분균명현고우TLR2단극륭항체조,차이균유통계학의의(t=4.183、4.954、13.506,균P＜0.05);TLR2단극륭항체조각막식존활시간위15.5d,95％가신구간(CI)위14.9～16.1,단순모형조각막식편존활시간위9.5d,95％ CI위8.7～10.3,2개조간차이유통계학의의(Z=12.728,P=0.001).각막조직병이학검사현시,단순모형조대서각막기질층수종명현,각막조직중가견대량염성세포침윤화신생혈관관강형성,이재TLR2단극륭항체조중부견소량염성세포화신생혈관.결론 TLR2단극륭항체능감경각막이식술후적염증반응,연장각막식편적존활시간.
Background The effects of Toll-like receptor 2 (TLR2) in grafting-related immune diseases have attracted more and more attention.Blocking TLR2 signal pathway can extend the survival time of heart and kidney grafts.However, the effects of anti-TLR2 monoclonal antibody on corneal graft have not been confirmed.Objective This study was to investigate the influence of anti-TLR2 monoclonal antibody on corneal graft survival in the rats received penetrating keratoplasty (PKP).Methods Allograft corneal transplantation was performed on the right eyes of 24 SPF female Wistar rats to establish PKP models,with 12 SD rats as donors.The model eyes were randomized into the TLR2 monoclonal antibody group and the model group.Anti-TLR2 monoclonal antibody of 15 μg/30 μl was subconjunctivally injected on day 0,2,4,6 and 8 following the modeling in the TLR2 monoclonal antibody group,and equal amount of normal saline was injected in the same way in the model group.The edema,transparency and neovascularization were observed under the slit lamp microscope after surgery, and rejection index (RI) was scored based on the criteria of Holland.Corneal tissue sections of the rats were prepared for the histopathological examination on day 9 and 15 after operation.The research protocol was approved by the Southern Medical University Ethics Committee.Results Mild corneal edema was found in the two groups 1-4 days after operation.A lot of new blood vessels, edema and opacification of corneas were seen in the model group 9-14 days after operation,but in the TLR2 monoclonal antibody group,corneal opacification was found 15 days after operation.The RI scores were significantly higher in the model group than those in the TLR2 monoclonal antibody group 5,9,15 days after operation (t=4.183,4.954,13.506;all at P＜0.05).The survival time in the TLR2 monoclonal antibody group was 15.5 days,with the 95％ confidence interval (CI) 14.9-16.1;while that in the model group was 9.5 days,with the 95％ CI 8.7-10.3, showing a significant difference between the two groups (Z =12.728,P =0.001).The corneal histopathological examination revealed that corneal stromal edema,infiltration of inflammatory cells and vascular lumen were more prominent 9 and 15 days after operation in the model group than those in the TLR2 monoclonal antibody group.Conclusions Anti-TLR2 monoclonal antibody can inhibit inflammatory response after allograft corneal transplantation and therefore extend the survival time of graft in rats.
