FK506抑制同种异体角膜缘移植术兔外周血T淋巴细胞CD25的表达

目的 探讨FK506滴眼液对同种异体角膜缘移植术后的免疫抑制作用。方法 新西兰兔48只48眼建立角膜缘缺陷症动物模型。随机分为FK506组、环孢霉素A(CsA)组及生理盐水对照组,每组16眼。1个月后实施同种异体角膜缘移植术,检测术前及术后1－8周兔外周血T淋巴细胞上CD25的表达。结果 各组表达CD25的外周血T淋巴细胞数均有升高。对照组较其他两组高;而CsA组显著高于FK506组(P＜0. 05)。 结论 FK506滴眼液可抑制同种异体角膜缘移植术后T淋巴细胞上CD25的表达,其免疫抑制效果优于CsA。     

FK506滴眼液抑制兔同种异体角膜缘移植排斥反应的免疫病理学研究

目的 :观察同种异体角膜缘移植术后排斥反应的免疫病理学变化 ,探讨FK5 0 6滴眼液的免疫抑制作用。方法 :新西兰兔 48只 48眼建立角膜缘缺陷症动物模型。随机分为PK 5 0 6组、环孢霉素A组 (CsA )及生理盐水对照组 ,每组16眼。 1月后实施同种异体角膜缘移植术。应用免疫病理学方法检测术后 4、 8、 10周角膜缘植片CD2 5的表达和淋巴细胞浸润。结果 :术后 4、 8、 10周三组角膜缘植片CD2 5的表达和淋巴细胞浸润均有升高 ,对照组较FK5 0 6组和CsA组高 (P <0 0 5 ) ,CsA组显著高于FK5 0 6组 (P <0 0 5 )。结论 :PK 5 0 6滴眼液可抑制同种异体角膜缘移植术后植片CD2 5的表达 ,其免疫抑制作用优于CsA。     

CsA滴眼液防治角膜缘移植排斥反应的实验研究

目的 探讨环孢霉素A(CsA)滴眼液对同种异体角膜缘移植术后的免疫抑制作用。方法 32只新西兰白兔随机分成CsA组和生理盐水对照组,每组16只。建立右眼角膜缘缺陷症动物模型,1月后实施同种异体角膜缘移植术,术后分别给予1％CsA滴眼液及生理盐水治疗,共4周。移植术后每日观察植片存活、角膜新生血管和上皮再生情况,共8周;术前1日及术后1～8周动态监测外周血T淋巴细胞CD25的表达。结果 对照组先于CsA组出现上皮排斥反应(P<0.05)。CsA组外周血T淋巴细胞CD25表达低于对照组(P<0.05)。结论 同种异体角膜缘移植术后早期应用CsA滴眼液可抑制外周血T淋巴细胞CD25的表达从而抑制排斥反应发生。     

FK506眼液防治同种异体角膜缘移植免疫排斥反应的实验研究

目的　探索同种异体角膜缘移植排斥反应规律及FK5 0 6眼液的免疫抑制作用。方法4 8只新西兰白兔随机均分成FK5 0 6组、环孢素A组及非治疗组 ,建立右眼角膜缘缺陷症动物模型 ,1个月后行同种异体角膜缘移植术 ,术后分别用 0 5 %FK5 0 6眼液、1%环孢素A眼液及生理盐水滴眼4周。角膜缘移植术前 1d ,术后 2、4周分别进行角膜中央印迹细胞学检查 ;术前 1d及术后 1～ 8周动态监测外周血T淋巴细胞CD2 5的表达 ;移植术后每日观察植片存活、角膜新生血管和上皮再生情况 ,共 10周 ;分别取术后第 4、8及 10周的角膜缘植片观测淋巴细胞浸润及T淋巴细胞上CD2 5的表达。结果　角膜缘创伤后 1个月 ,所有兔眼角膜中央印迹细胞学检查均为结膜表型 ;同种异体角膜缘移植术后 4周 ,FK5 0 6组和环孢素A组角膜中央保持角膜细胞表型 ,非治疗组呈结膜细胞表型。非治疗组最早出现上皮排斥反应 ,FK5 0 6与环孢素A均可抑制排斥反应的发生 ,停药后环孢素A组先于FK5 0 6组出现上皮排斥反应 (P <0 0 5 )。FK5 0 6组外周血、角膜缘植片T淋巴细胞CD2 5表达及角膜缘植片中淋巴细胞浸润程度低于环孢素A组 ,两组均低于非治疗组 (P <0 0 5 )。结论　同种异体角膜缘移植术后早期应用 0 5 %FK5 0 6眼液滴眼可抑制外周血及植片T淋巴细胞CD2 5     

表面应用FK506对全角膜移植免疫排斥的预防和治疗作用

目的探讨FK506（他克莫司）滴眼液对全角膜移植免疫排斥的预防和治疗作用。方法对全角膜和亚全角膜受损毁的50眼进行全角膜移植。术后病人分两组,0.05%FK506滴眼液联合0.1%地塞米松滴眼液的FK506治疗组和1%环孢霉素A（cyclosporine A,CSA）滴眼液联合0.1%地塞米松滴眼液的CSA治疗组,比较它们预防和治疗移植排斥的疗效。结果平均随访18.6m±6.2m（6～27m）。总的眼球保存率96%,角膜植片总透明率42.9%。术后,23眼视力超过0.02,3眼视力超过0.3,最好视力1.2。FK506组与CSA组相比,明显降低和推迟移植排斥（P=0.005和P=0.036）。FK506组最终视力和术后植片透明率均高于CSA组（P=0.001和P=0.017）。FK506组术后6m和12m的植片透明率分别是74.1%和60.5%,而CSA组分别是42.1%和24.1%。结论通过早期联合表面应用FK506和皮质类固醇,后期单独表面应用FK506,全角膜移植不仅可以挽救角膜和眼前段严重受损的眼球,而且使多数病人获得较好的视力。     

FK506缓释系统前房植入抑制兔高危角膜移植术后的免疫排斥反应

目的探讨前房植入FK506药物缓释系统（DDS）对兔高危角膜移植术后免疫排斥反应的抑制作用和FK506房水药物浓度与免疫排斥反应的关系。方法107只新西兰白兔中随机数字法选取73只兔进行角膜新生血管化模型的制作,其中68只兔作为受体成功建立高危角膜移植动物模型,随机数字法分为对照组、空白DDS前房植入组、环孢素A（CsA）DDS前房植入组（含CsA 1mg）、0．1％FK506眼液滴眼组及FK506 DDS前房植入组（含FK5060．5mg）。角膜移植术后观察各组角膜植片排斥发生的时间,移植术后1周取各组实验兔眼房水和静脉血进行FK506药物浓度检测。0．1％FKS06眼液滴眼组和FKS06 DDS前房植入组在移植术后的不同时间点抽取实验兔眼房水和静脉血,进行FK506药物浓度的检测。观察各组兔移植术后4周和观察期结束时角膜植片的病理变化,同时应用原位杂交的方法检测各组角膜植片内白细胞介素2受体a（IL-2Bot）、单核细胞趋化蛋白1（MCP-1）、Fas及FasL mRNA的表达。结果FK506 DDS前房植入组角膜植片存活时间超过180d,明显优于其他各组（F=926．37,P=0．0000）,其房水和角膜组织中的FK506药物浓度明显高于FKS06眼液滴眼组（T=21．00,P=0．0022）。FKS06 DDS前房植入组在术后24周内均能在房水中检测出FK506。术后4周对照组和空白DDS前房植入组有大量的炎性细胞浸润,并有明显的IL．2Bet和MCP-1mRNA的表达,而CsADDS前房植入组、FK506眼液滴眼组及FK506 DDS植入组角膜未见明显的炎性细胞浸润,未见IL-2Pux和MCP-1mRNA的表达。各组均未见明显的Fas和FasL mRNA的表达。结论前房植入FK506 DDS可有效地抑制高危角膜移植术后免疫排斥反应的发生,房水中较高的FK506药物浓度是防治术后发生免疫排斥反应的重要因素。     

FK506滴眼液联合角膜移植术治疗复发性蚕蚀性角膜溃疡

目的：观察FK506滴眼液联合角膜移植术治疗复发性蚕蚀性角膜溃疡的疗效。方法：采用FK506滴眼液联合角膜移植术治疗复发性吞蚀性角膜溃疡患者9例（15只眼），对其中角膜溃疡＜2个象限角膜缘的2只眼，采用局部滴用0．1％，FD506滴眼液：角膜溃疡＞2个象限角膜缘的13只眼中，12只眼行球结膜切除及板层角膜移植术，1只眼因角膜溃疡穿孔，行穿透性角膜移植术，待术眼角膜上皮愈合后，局部滴用0．1％，FK506滴眼液，观察其疗效，同时对术中获取的角膜，结膜组织及房水，用酶联免疫分析法测定0．1％，FK506滴眼液在角膜，结膜组织及房水中的含量，对照组为板层角膜移植术联合0．1％地塞米松滴眼液治疗的12例复发性吞蚀性角膜溃疡患者。结果：滴用0．1％ FK506滴眼液后，角膜和结膜组织中FK506的含量为30－350ng/g，房水中未检测出FK506。9例（15只眼）吞蚀性角膜溃疡患者滴用0．1％，FD506滴眼液或联合角膜移植术治疗后，角膜溃疡均愈合，随访观察12－17个月，角膜溃疡无复发。视力提高＞2行者5只眼，对照组中，有7只眼的角膜溃疡复发，结论：局部应用0．1％ FK506滴眼液联合角膜移植术是治疗复发性吞蚀性角膜溃疡的有效方法。     

山茱萸总甙滴眼液防治角膜移植免疫排斥反应的实验研究

目的：观察山茱萸总甙（COG)滴眼液局部应用对角膜移植免疫排斥反应的影响。方法：建立封闭群大鼠角膜移植模型，随机分组：A、B、C、D组为Wister—SD组问同种异体角膜移植，Wistar为受体，SD为供体，其中A组为空白对照组、B组为0．5％COG滴眼液组、C组为1％COG滴眼液组，D组为2％COG滴眼液组，E组为Wistar—Wistar对照组，即Wistar大鼠间同种异体移植组。用裂隙灯显微镜记录及比较各组：角膜植片透明度、水肿度、新生血管度、移植排斥指数(RI)及角膜排斥发生时间。结果：术后各组移植排斥指数及发生角膜移植排斥时间，A组与B组、A组与C组、A组与D组、B组与C组、C组与D组比较均有显著性差异，P＜0．01；B组与D组之间比较P＞0．05，无显著性差异。结论：COG滴眼液能有效地防治角膜移植免疫排斥反应，1％COG滴眼液更有效。     

局部联合应用雷帕霉素及环孢霉素A在大鼠角膜移植排斥反应中的作用

目的:评价雷帕霉素滴眼液与环孢霉素A滴眼液联合应用对大鼠角膜移植免疫排斥反应的抑制作用。方法:建立大鼠穿透性角膜移植动物模型,以SD大鼠为受体,Wistar大鼠为供体,分为A,B,C,D4组。手术后2d术眼分别滴用滴眼液基质100μL,2g/L雷帕霉素滴眼液100μL,10g/L环孢霉素A滴眼液100μL,2g/L雷帕霉素滴眼液50μL 10g/L环孢霉素A滴眼液50μL,4次/d,连续用药至排斥反应发生。对角膜植片进行临床观察,记录植片存活时间,并进行组织学检查。结果:各治疗组显著延长角膜植片存活时间(P<0.01),联合用药的D组效果更佳,优于各单药物治疗组B组和C组(P<0.05,P<0.01)。组织学发现,术后14d各治疗组炎细胞浸润、新生血管形成及水肿程度明显轻于对照组。结论:局部联合应用雷帕霉素滴眼液与环孢霉素A滴眼液能显著延长角膜植片存活时间,抑制大鼠角膜移植排斥反应,具有协同作用。     

来氟米特活性代谢物A771726对大鼠角膜移植排斥反应的影响

目的探讨来氟米特活性代谢物A771726对大鼠角膜移植排斥反应的抑制作用。方法建立SD-Wistar大鼠同种异体穿透角膜移植模型,随机分组。A组为空白对照组;B、C、D组分别为0．5％、1．0％及2．0％A771726滴眼液组;E组为Wistar大鼠自体移植对照组。术后比较各组角膜植片排斥指数（RI）及植片存活时间,并对植片进行组织学及免疫组织化学染色观察。结果A组角膜植片存活时间为（9．38±2．26）d,B组（10．13±2．41）d,C组（17．57±1．72）d,D组（17．50±2．14）d,E组（〉28．00）d。A组、B组分别与C组、D组植片存活时间差异有统计学意义（P〈0．01）。移植术后10d,A组、B组角膜植片发生排斥反应,植片高表达IFN-γ及ICAM-1;术后20d,C组、D组植片发生排斥反应,植片IFN-γ及ICAM-1表达增高。结论局部应用A771726滴眼液能有效抑制角膜移植排斥反应。     

FK-506抑制同种异体角膜缘移植术后免疫排斥反应的研究

目的：探讨ＦＫ－５０６抑制角膜缘干细胞移植术后免疫排斥反应的临床可行性与有效性。方法：应用前瞻性评估研究方法,将角膜缘移植术后病例６４只眼按随机原则分投药组及对照组各半,投药组应用０．５％ＦＫ－５０６滴眼液,对照组应用１％ＣｓＡ滴眼液。平均随访期半年,以术后角膜缘植片新生血管、水肿、混浊及溶解程度为临床主要评估指标。结果：随访期内在新生血管指数、上皮排斥发生率、移植排斥指数及假性胬肉指数四项指标投     

Limbal autograft and allograft transplantations in patients with corneal burns

AIM: To investigate and compare the surgical outcomes of limbal autograft and limbal allograft transplantations in patients with corneal burns. METHODS: In total, 20 patients (n=22 eyes) with chemical burn and two patients (n=2 eyes) with thermal burn were included in this study. Limbal autograft or limbal allograft transplantation surgery was performed in all patients. HLA-typing was tested before allograft surgeries. Limbal allografting was performed in all eyes using donor tissue from live relatives. Systemic cyclosporine A was administered for immunosuppression. RESULTS: The corneal surface was successfully reconstructed in all eyes (100%) after limbal autografting, two eyes required additional amniotic membrane transplantation and one eye required allografting. The mean follow-up period for limbal autografts was 13.9 +/- 7.0 months. Limbal allografting failed to reduce corneal vascularity and opacification in five (55.6%) eyes and was successful only in four (44.4%) eyes (mean follow-up 16.2 +/- 11.2 months) (P=0.002). In all, 15 eyes undergoing limbal autografting completed re-epithelialization of the cornea at a mean of 35.6 +/- 60.2 days. The mean epithelial healing time in nine eyes undergoing limbal allografting was 13.0 +/- 7.3 days (P=0.525). After limbal autografting, functional vision (> or =1/10) was attained in 12 (80%) eyes. Only one eye (11.1%) achieved functional vision after limbal allografting (P=0.036). Penetrating keratoplasty was performed in three patients following limbal allografting. No cyclosporine-associated side effects were observed. CONCLUSIONS: Limbal autograft transplantation is an effective and safe procedure for unilateral corneal burns. It seems that limbal allograft transplantation is better combined with penetrating keratoplasty for a better visual outcome and higher graft survival rate. Systemic immunosuppression seems to be necessary for limbal allografts even in the presence of HLA-matched donor tissues.     

Efficacy of systemic cyclosporine A and amniotic membrane on rabbit conjunctival limbal allograft rejection

PURPOSE: To evaluate the effect of intramuscular cyclosporine A (CsA) and amniotic membrane (AM) on conjunctival limbal allograft survival in a rabbit model. METHODS: Eighty-two female rabbits (59 New Zealand white rabbits, 23 Dutch pigmented rabbits) were used. The New Zealand white rabbits were divided into 4 treatment groups: group 1 (n=13), conjunctival limbal autograft transplantation; group 2 (n=12), conjunctival limbal allograft transplantation without additional treatment; group 3 (n=18), conjunctival limbal allograft transplantation and human AM; and group 4 (n=16), conjunctival limbal allograft transplantation and systemic CsA (10 mg/kg/day intramuscularly). The 23 Dutch pigmented rabbits were used as limbal stem cell allograft donors. The rejection index, the mean survival time, and the rejection rates were calculated for each group. RESULTS: After 28 days of follow-up, there were no episodes of limbal rejection in groups 1 and 4, whereas the rejection rate was 100% in groups 2 and 3. There was no significant difference in mean survival time of the rejected grafts between groups 2 and 3. CONCLUSIONS: A model of rejection of conjunctival limbal transplantation was developed in the rabbit. Intramuscularly injected CsA effectively prevents limbal allograft rejection. Human AM is not useful for this purpose.     

Preliminary study of the effect of FK506 nanospheric-suspension eye drops on rejection of penetrating keratoplasty.

OBJECTIVE: The aim of this study was to investigate the effect of a topical FK506 nanospheric suspension in a rat model of penetrating keratoplasty. METHODS: FK506 nanospheres were prepared by using a biodegradable poly (lactic-co-glycolic acid) copolymer (PLGA). Its distribution in the eye and blood after a single instillation was examined in rabbits. Sprague-Dawley (SD) rats received corneal heterografts and were topically treated with phosphate-buffered saline (PBS), PLGA, FK-506 0.01% (nanospheres), or dexamethasone 0.05% solutions twice a day for 28 days. Rejection index and graft-survival time were recorded and compared between the four groups. Three grafts were collected at different time points for immunohistochemical studies. RESULTS: In the cornea, the FK-506 concentration reached its peak within 1 h of a single eye-drop instillation and then decreased by half (1667.85 +/- 611.87 ng/g) at 8 h. FK-506 cannot be detected in rabbit blood. There were significant differences in the graft-survival time between the FK-506 nanosphere group (15.09 +/- 4.81 days) and the other three groups [PBS (7.90 +/- 1.20, t = -4.594, P < 0.001), PLGA (8.44 +/- 0.88, t = - 4.074, P = 0.001) and dexamethasone (10.44 +/- 1.42, t = -2.790, P = 0.012)]. The rejected corneas in the FK506 nanosphere group showed significantly fewer CD4, CD8, CD68, CD79, vascular endothelial growth factor, ICAM, and tumor growth factor-beta(1)-positive cells than those in the other groups. CONCLUSIONS: FK506 0.01% nanospheric-suspension eye drops delayed the occurrence of corneal allograft rejection and prolonged allograft survival time. The FK506 nanospheres may be valuable in suppressing corneal graft rejection.     

Limbal allografting from related live donors for corneal surface reconstruction

OBJECTIVE: To report the results of limbal allograft transplantation, from human leukocyte antigen (HLA)-matched and -unmatched related live donors, in patients with ocular surface disease due to chemical burns and Stevens-Johnson syndrome. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: Eight patients (nine eyes) with severe chemical burns (n = 7 eyes) and Stevens-Johnson syndrome (n = 2 eyes). INTERVENTION: Recipient eyes were treated with excision of cicatricial tissues. Transplantation of superior and inferior limbal grafts was performed from related live HLA-matched (n = 7) and -unmatched donors (n = 2). Systemic cyclosporine was not used in any of the recipients. MAIN OUTCOME MEASURES: Reconstruction of corneal surface epithelium, restoration of avascularity, increase in ocular comfort, and improvement in visual acuity. RESULTS: With a mean observation period of 17.2 months, phenotypically corneal epithelium, decreased vascularization of the corneal surface, and improved ocular comfort were seen in seven (77.8%) eyes. In all seven eyes, gradual recurrence of peripheral corneal vascularization occurred during the follow-up period. Features of graft rejection developed in three (42.9%) of these seven eyes. In two eyes, limbal transplantation from HLA-unmatched donors failed to reconstitute the corneal surface. Limbal allograft transplantation resulted in visual acuity of 20/400 or greater in only two (22.2%) eyes at last follow-up. Corneal grafts performed 7 and 16 months after successful limbal transplantation in two eyes developed recurrent epithelial breakdown and superficial corneal scarring. None of the donor eyes in this study had any complication. CONCLUSION: Transplantation of limbal tissue from related live donors successfully reconstructs the corneal surface in HLA-matched recipients. Recurrence of vascularization on long-term follow-up probably results from inadequate stem cell transfer, immune-mediated stem cell damage, or both. Limbal allografting is best performed by transplanting the entire limbus from a cadaveric donor eye with systemic immunosuppression of the recipient, even if the donor is HLA-compatible.     

环孢素A不同给药方式抑制兔眼穿透角膜移植术后免疫排斥反应的研究

背景环孢素A（CsA）是治疗角膜移植免疫排斥反应的主要药物,但合适的药物剂型和给药途径对提高药物利用度有重要意义。目的探讨CsA缓释微球结膜下给药、前房给药及CsA滴眼液局部点眼途径抑制兔眼穿透角膜移植术（PKP）后的排斥反应。方法健康清洁级成年新西兰白兔60只60只眼作为受体,健康清洁级青紫蓝兔30只60只眼作为供体。将受体兔按随机数字表法随机分为空白对照组、结膜下给药组、结膜下对照组、前房给药组、前房对照组和CsA滴眼液组,每组10只实验兔。所有实验兔行PKP。术毕结膜下给药组和前房给药组兔眼以相应的给药途径注射12g／LCsA缓释微球悬液0．1ml,结膜下对照组和前房对照组采取相应的给药途径注射空白微球悬液0．1ml,CsA滴眼液组每日应用质量分数1％（10g／L）CsA滴眼液点眼,空白对照组PKP术后不给予CsA药物。术后裂隙灯显微镜下定期观察各组术眼角膜植片的透明度、水肿情况、新生血管生长情况等,并计算术眼的排斥反应指数（RI）。分别于术前,术后3d,术后1、2、3周和术后1、2、3个月用Tono—pen眼压计测量眼压;分别于术后1个月、3个月获取各组植片行常规组织病理学检查。结果术后各组兔眼眼压较术前均明显下降,手术前后兔眼眼压的总体比较差异有统计学意义（F目目：29．210,P=0．000）;同一时间点各组兔眼眼压比较差异无统计学意义（F捆=0．254,P=0．938）。空白对照组、结膜下对照组及前房对照组于术后2～3周出现不同程度的角膜植片混浊和新生血管,术后4周时植片混浊加重,R1分别为8．60±1．52、8．60±0．55和8．80±0．84;结膜下给药组、前房给药组及CsA滴眼液组于术后3周时出现角膜新生血管,但未发现植片混浊,R1分别为4．40＋0．89、3．20±0．84和3．00±0．71,均明显低于空白对照组、结膜下对照组和前房对照组,差异均有统计学意义（P〈0．05）。其中前房给药组兔眼术后前房有轻度炎症反应,随时间延长逐渐减轻,至术后3个月时}肖失。组织病理学检查可见,空白对照组、结膜下对照组、前房对照组术眼角膜植片均明显增厚,有大量炎性细胞浸润和新生血管长人;而结膜下给药组、前房给药组和CsA滴眼液组植片的炎性细胞浸润明显减轻,新生血管减少。结论CsA缓释微球经不同途径给药均可抑制兔眼角膜移植术后的排斥反应,其中经前房给药途径的整体疗效较经结膜下给药途径更佳。