中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
10期
902-907
,共6页
解孝锋%田庆梅%毕宏生%温莹%张竖%郭爱莲
解孝鋒%田慶梅%畢宏生%溫瑩%張豎%郭愛蓮
해효봉%전경매%필굉생%온형%장수%곽애련
角膜移植术/高危%免疫排斥反应%祖国传统医学%中医药
角膜移植術/高危%免疫排斥反應%祖國傳統醫學%中醫藥
각막이식술/고위%면역배척반응%조국전통의학%중의약
Corneal transplantation/high-risk corneal transplantation%Immunne rejection%Traditional Chinese medicine%Chinese medicine
背景 高危角膜移植患者虽然常规应用免疫抑制性药物,但角膜移植片的排斥率仍很高.近年来传统中药在角膜移植中的作用逐渐引起关注,黑睛排斥汤对大鼠高危角膜移植免疫排斥反应的防治作用需要进一步研究. 目的 探讨黑睛排斥汤对大鼠高危角膜移植免疫排斥反应的抑制作用. 方法 16只正常雌性SD大鼠角膜作为供体,32只雌性Wistar大鼠作为受体,用角膜缝线法诱导角膜新生血管(CNV),建立大鼠高危角膜移植模型,然后行封闭群大鼠SD-Wistar组间同种异体全板层角膜移植.采用随机数字表法将大鼠随机分为4个组,包括模型对照组(生理盐水灌服)、CsA组[10 mg/(kg·d) CsA灌服]、黑睛排斥汤组[4g/(kg·d)黑睛排斥汤灌服]和联合用药组[10 mg/(kg·d)CsA+4 g/(kg·d)黑睛排斥汤灌服],各组大鼠均于同种异体角膜移植后4d开始灌服相应药物,于术后第4天起裂隙灯显微镜下观察眼前节反应,每隔2天观察1次,连续观察30 d.参考Larkin等的标准对角膜混浊程度、水肿程度及新生血管程度进行评分,记录移植排斥指数(RI)并观察植片的存活状况,对角膜进行常规组织病理学检查,采用流式细胞分析法对受体角膜局部和全身免疫状态进行检测,对4个组大鼠的各种检测指标进行比较.结果 模型对照组大鼠角膜移植术后10d左右发生排斥反应,CsA组和黑睛排斥汤组大鼠均于术后12 ～ 13 d发生排斥反应,而联合用药组大鼠于术后22 d发生排斥反应.与模型对照组比较,CsA组、黑睛排斥汤组和联合用药组大鼠角膜移植后角膜混浊程度评分、角膜水肿评分和新生血管评分均明显下降,差异均有统计学意义(P＜0.05),且联合用药组上述各指标的评分均明显低于CsA组和黑睛排斥汤组,差异均有统计学意义(P＜0.05),而CsA组与黑睛排斥汤组的各种评分差异均无统计学意义(P＞0.05).模型对照组角膜移植片存活时间平均为(10.38±1.69)d,联合用药组平均为(22.50±3.07)d,差异有统计学意义(t=-9.790,P=0.000).组织病理学检查表明,术后15d联合用药组大鼠角膜基质中淋巴细胞浸润及新生血管评分少于CsA组和黑睛排斥汤组.流式细胞学检查证实,黑睛排斥汤组、CsA组和联合用药组大鼠角膜移植术后外周血中CD4+淋巴细胞计数及CD4+/CD8+比值均低于模型对照组,联合用药组CD4+淋巴细胞计数及CD4+/CD8+比值均低于CsA组和黑睛排斥汤组,差异均有统计学意义(P＜0.05).结论 黑睛排斥汤对高危角膜移植模型大鼠术后角膜排斥反应有抑制作用,其疗效与0.1％CsA接近,二者联合用药有协同作用.
揹景 高危角膜移植患者雖然常規應用免疫抑製性藥物,但角膜移植片的排斥率仍很高.近年來傳統中藥在角膜移植中的作用逐漸引起關註,黑睛排斥湯對大鼠高危角膜移植免疫排斥反應的防治作用需要進一步研究. 目的 探討黑睛排斥湯對大鼠高危角膜移植免疫排斥反應的抑製作用. 方法 16隻正常雌性SD大鼠角膜作為供體,32隻雌性Wistar大鼠作為受體,用角膜縫線法誘導角膜新生血管(CNV),建立大鼠高危角膜移植模型,然後行封閉群大鼠SD-Wistar組間同種異體全闆層角膜移植.採用隨機數字錶法將大鼠隨機分為4箇組,包括模型對照組(生理鹽水灌服)、CsA組[10 mg/(kg·d) CsA灌服]、黑睛排斥湯組[4g/(kg·d)黑睛排斥湯灌服]和聯閤用藥組[10 mg/(kg·d)CsA+4 g/(kg·d)黑睛排斥湯灌服],各組大鼠均于同種異體角膜移植後4d開始灌服相應藥物,于術後第4天起裂隙燈顯微鏡下觀察眼前節反應,每隔2天觀察1次,連續觀察30 d.參攷Larkin等的標準對角膜混濁程度、水腫程度及新生血管程度進行評分,記錄移植排斥指數(RI)併觀察植片的存活狀況,對角膜進行常規組織病理學檢查,採用流式細胞分析法對受體角膜跼部和全身免疫狀態進行檢測,對4箇組大鼠的各種檢測指標進行比較.結果 模型對照組大鼠角膜移植術後10d左右髮生排斥反應,CsA組和黑睛排斥湯組大鼠均于術後12 ～ 13 d髮生排斥反應,而聯閤用藥組大鼠于術後22 d髮生排斥反應.與模型對照組比較,CsA組、黑睛排斥湯組和聯閤用藥組大鼠角膜移植後角膜混濁程度評分、角膜水腫評分和新生血管評分均明顯下降,差異均有統計學意義(P＜0.05),且聯閤用藥組上述各指標的評分均明顯低于CsA組和黑睛排斥湯組,差異均有統計學意義(P＜0.05),而CsA組與黑睛排斥湯組的各種評分差異均無統計學意義(P＞0.05).模型對照組角膜移植片存活時間平均為(10.38±1.69)d,聯閤用藥組平均為(22.50±3.07)d,差異有統計學意義(t=-9.790,P=0.000).組織病理學檢查錶明,術後15d聯閤用藥組大鼠角膜基質中淋巴細胞浸潤及新生血管評分少于CsA組和黑睛排斥湯組.流式細胞學檢查證實,黑睛排斥湯組、CsA組和聯閤用藥組大鼠角膜移植術後外週血中CD4+淋巴細胞計數及CD4+/CD8+比值均低于模型對照組,聯閤用藥組CD4+淋巴細胞計數及CD4+/CD8+比值均低于CsA組和黑睛排斥湯組,差異均有統計學意義(P＜0.05).結論 黑睛排斥湯對高危角膜移植模型大鼠術後角膜排斥反應有抑製作用,其療效與0.1％CsA接近,二者聯閤用藥有協同作用.
배경 고위각막이식환자수연상규응용면역억제성약물,단각막이식편적배척솔잉흔고.근년래전통중약재각막이식중적작용축점인기관주,흑정배척탕대대서고위각막이식면역배척반응적방치작용수요진일보연구. 목적 탐토흑정배척탕대대서고위각막이식면역배척반응적억제작용. 방법 16지정상자성SD대서각막작위공체,32지자성Wistar대서작위수체,용각막봉선법유도각막신생혈관(CNV),건립대서고위각막이식모형,연후행봉폐군대서SD-Wistar조간동충이체전판층각막이식.채용수궤수자표법장대서수궤분위4개조,포괄모형대조조(생리염수관복)、CsA조[10 mg/(kg·d) CsA관복]、흑정배척탕조[4g/(kg·d)흑정배척탕관복]화연합용약조[10 mg/(kg·d)CsA+4 g/(kg·d)흑정배척탕관복],각조대서균우동충이체각막이식후4d개시관복상응약물,우술후제4천기렬극등현미경하관찰안전절반응,매격2천관찰1차,련속관찰30 d.삼고Larkin등적표준대각막혼탁정도、수종정도급신생혈관정도진행평분,기록이식배척지수(RI)병관찰식편적존활상황,대각막진행상규조직병이학검사,채용류식세포분석법대수체각막국부화전신면역상태진행검측,대4개조대서적각충검측지표진행비교.결과 모형대조조대서각막이식술후10d좌우발생배척반응,CsA조화흑정배척탕조대서균우술후12 ～ 13 d발생배척반응,이연합용약조대서우술후22 d발생배척반응.여모형대조조비교,CsA조、흑정배척탕조화연합용약조대서각막이식후각막혼탁정도평분、각막수종평분화신생혈관평분균명현하강,차이균유통계학의의(P＜0.05),차연합용약조상술각지표적평분균명현저우CsA조화흑정배척탕조,차이균유통계학의의(P＜0.05),이CsA조여흑정배척탕조적각충평분차이균무통계학의의(P＞0.05).모형대조조각막이식편존활시간평균위(10.38±1.69)d,연합용약조평균위(22.50±3.07)d,차이유통계학의의(t=-9.790,P=0.000).조직병이학검사표명,술후15d연합용약조대서각막기질중림파세포침윤급신생혈관평분소우CsA조화흑정배척탕조.류식세포학검사증실,흑정배척탕조、CsA조화연합용약조대서각막이식술후외주혈중CD4+림파세포계수급CD4+/CD8+비치균저우모형대조조,연합용약조CD4+림파세포계수급CD4+/CD8+비치균저우CsA조화흑정배척탕조,차이균유통계학의의(P＜0.05).결론 흑정배척탕대고위각막이식모형대서술후각막배척반응유억제작용,기료효여0.1％CsA접근,이자연합용약유협동작용.
The rate of corneal graft rejection is still high for high-risk keratoplasty although immune suppression drug is routinely used.The role of traditional Chinese medicine in corneal transplantation is concerned gradually.Heijingpaichitang on the prevention and treatment of rats with high-risk corneal allograft rejection needs further study.Objective This study was to investigate the inhibitory effect of heijingpaichitang on high-risk corneal transplantation immune rejection in rats.Methods Sixteen female SD rats were used as the donors and 32female Wistar rats were served as recipients.The high-risk corneal trasplantation models were established by corneal suture in 32 Wistar rats,and then homogeneity variant SD-Wistar corneal transplantation was performed.The recipients were randomized into model control group,cyclosporinc A (CsA)group,heijingpaichitang group and CsA +heijingpaichitang group.CsA,heijingpaichitang and CsA + heijingpaichitang was orally administered 4 days after operation once per day for 15 days,and normal saline solution was used at the same way in the model control group.Ocular anterior segment reaction was examined under the slit lamp and corneal opacification,edema and neovasculation were scored based on Larkin' s criteria.Rejection index of the corneal graft was recorded and the graft survival time was calculated.The pathological examination of the corneal graft was carried out in all rats,and the inflammatory cells in the corneas and CD4+ cells in the periphery blood were assayed using flow cytometry.The use of the animals complied with ARVO Statement.Results Corneal graft rejection occurred in 10 days after operation in the model control group,12-13 days in the CsA group and heijingpaichitang group and 22 days in the CsA +heijingpaichitang group.Compared with model control group,the scores of the corneal opacification,corneal edema and neovascularization were significantly lower in the CsA group,heijingpaichitang group and CsA+heijingpaichitang group (P＜0.05),and all the scores were declined in the CsA+ heijingpaichitang group compared with CsA group and heijingpaichitang group(P＜0.01),but no significant differences were seen in the scores between the CsA group and heijingpaichitang group(P＞0.05).The mean survival time of grafts was (10.38 ±1.69)days in the model control group,(22.50 ± 3.07) days in the CsA + heijingpaichitang group,with the significant difference (t =-9.790,P =0.000).The pathological examination of graft showed that the lymphocytes and new blood vessels were less in the CsA+heijingpaichitang group compared with CsA group and heijingpaichitang group 15 days after operation.Flow cytometry verified that the number of lymphocytes in graft,CD4+cells and CD4+/CD8+ in periphery blood were significantly lower in the heijingpaichitang group,CsA group and CsA+heijingpaichitang group compared with model control group (P＜0.05).Conclusions Heijingpaichitang can inhibit immune rejection to certain extent in high-risk corneal transplantation rat and has a similar effect to 0.1％ CsA.Heijingpaichitang and 0.1％ CsA have a synergistic effect.
