CAJ | 학술논문

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12 g/L CsA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％(10 g/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l、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时间=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分组=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,RI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,RI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个月时消失.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入；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. 揹景環孢素A(CsA)是治療角膜移植免疫排斥反應的主要藥物,但閤適的藥物劑型和給藥途徑對提高藥物利用度有重要意義. 目的探討CsA緩釋微毬結膜下給藥、前房給藥及CsA滴眼液跼部點眼途徑抑製兔眼穿透角膜移植術(PKP)後的排斥反應.方法健康清潔級成年新西蘭白兔60隻60隻眼作為受體,健康清潔級青紫藍兔30隻60隻眼作為供體.將受體兔按隨機數字錶法隨機分為空白對照組、結膜下給藥組、結膜下對照組、前房給藥組、前房對照組和CsA滴眼液組,每組10隻實驗兔.所有實驗兔行PKP.術畢結膜下給藥組和前房給藥組兔眼以相應的給藥途徑註射12 g/L CsA緩釋微毬懸液0.1ml,結膜下對照組和前房對照組採取相應的給藥途徑註射空白微毬懸液0.1ml,CsA滴眼液組每日應用質量分數1％(10 g/L) CsA滴眼液點眼,空白對照組PKP術後不給予CsA藥物.術後裂隙燈顯微鏡下定期觀察各組術眼角膜植片的透明度、水腫情況、新生血管生長情況等,併計算術眼的排斥反應指數(RI).分彆于術前,術後3d,術後l、2、3週和術後1、2、3箇月用Tono-pen眼壓計測量眼壓；分彆于術後1箇月、3箇月穫取各組植片行常規組織病理學檢查.結果術後各組兔眼眼壓較術前均明顯下降,手術前後兔眼眼壓的總體比較差異有統計學意義(F時間=29.210,P=0.000)；同一時間點各組兔眼眼壓比較差異無統計學意義(F分組=0.254,P=0.938).空白對照組、結膜下對照組及前房對照組于術後2～3週齣現不同程度的角膜植片混濁和新生血管,術後4週時植片混濁加重,RI分彆為8.60±1.52、8.60±0.55和8.80±0.84;結膜下給藥組、前房給藥組及CsA滴眼液組于術後3週時齣現角膜新生血管,但未髮現植片混濁,RI分彆為4.40±0.89、3.20±0.84和3.00±0.71,均明顯低于空白對照組、結膜下對照組和前房對照組,差異均有統計學意義(P＜0.05).其中前房給藥組兔眼術後前房有輕度炎癥反應,隨時間延長逐漸減輕,至術後3箇月時消失.組織病理學檢查可見,空白對照組、結膜下對照組、前房對照組術眼角膜植片均明顯增厚,有大量炎性細胞浸潤和新生血管長入；而結膜下給藥組、前房給藥組和CsA滴眼液組植片的炎性細胞浸潤明顯減輕,新生血管減少. 結論 CsA緩釋微毬經不同途徑給藥均可抑製兔眼角膜移植術後的排斥反應,其中經前房給藥途徑的整體療效較經結膜下給藥途徑更佳.
배경 배포소A(CsA)시치료각막이식면역배척반응적주요약물,단합괄적약물제형화급약도경대제고약물이용도유중요의의. 목적 탐토CsA완석미구결막하급약、전방급약급CsA적안액국부점안도경억제토안천투각막이식술(PKP)후적배척반응.방법 건강청길급성년신서란백토60지60지안작위수체,건강청길급청자람토30지60지안작위공체.장수체토안수궤수자표법수궤분위공백대조조、결막하급약조、결막하대조조、전방급약조、전방대조조화CsA적안액조,매조10지실험토.소유실험토행PKP.술필결막하급약조화전방급약조토안이상응적급약도경주사12 g/L CsA완석미구현액0.1ml,결막하대조조화전방대조조채취상응적급약도경주사공백미구현액0.1ml,CsA적안액조매일응용질량분수1％(10 g/L) CsA적안액점안,공백대조조PKP술후불급여CsA약물.술후렬극등현미경하정기관찰각조술안각막식편적투명도、수종정황、신생혈관생장정황등,병계산술안적배척반응지수(RI).분별우술전,술후3d,술후l、2、3주화술후1、2、3개월용Tono-pen안압계측량안압；분별우술후1개월、3개월획취각조식편행상규조직병이학검사.결과 술후각조토안안압교술전균명현하강,수술전후토안안압적총체비교차이유통계학의의(F시간=29.210,P=0.000)；동일시간점각조토안안압비교차이무통계학의의(F분조=0.254,P=0.938).공백대조조、결막하대조조급전방대조조우술후2～3주출현불동정도적각막식편혼탁화신생혈관,술후4주시식편혼탁가중,RI분별위8.60±1.52、8.60±0.55화8.80±0.84;결막하급약조、전방급약조급CsA적안액조우술후3주시출현각막신생혈관,단미발현식편혼탁,RI분별위4.40±0.89、3.20±0.84화3.00±0.71,균명현저우공백대조조、결막하대조조화전방대조조,차이균유통계학의의(P＜0.05).기중전방급약조토안술후전방유경도염증반응,수시간연장축점감경,지술후3개월시소실.조직병이학검사가견,공백대조조、결막하대조조、전방대조조술안각막식편균명현증후,유대량염성세포침윤화신생혈관장입；이결막하급약조、전방급약조화CsA적안액조식편적염성세포침윤명현감경,신생혈관감소. 결론 CsA완석미구경불동도경급약균가억제토안각막이식술후적배척반응,기중경전방급약도경적정체료효교경결막하급약도경경가.
Background Cyclosporine A (CsA) is an effective drug to prevent rejection response after penetrating keratoplasty (PKP).Appropriate dosage forms and right administrating route is very important for improving the bioavailability of CsA.Objective This study was to investigate the preventing effect of CsA microspheres via subconjunctive and anterior chamber injection and CsA eye drops on immune rejection after PKP.Methods Sixty eyes of 60 clean adult New Zealand white rabbits served as receipts,and 60 eyes of 30 clean adult pigmented rabbits were used as the donors.The receipts were randomized into the blank control group (only PKP group),subconjunctival CsA injection group,subconjunctival vector injection group,anterior chamber CsA injection group,anterior chamber vector injection group,and 1％ CsA eye drops group.PKP was performed on the all rabbits,and then the CsA microsphere (0.1 ml,12 g/L) or blank microsphere (0.1 ml) were respectively administered in the corresponding groups.The corneal grafts were examined by slit lamp microscope regularly,and rejection index (RI)was calculated based on the corneal opacity,edema and neovascularization.The intraocular pressure (IOP) of operative eyes was measured by Tono-pen tonometer before operation,3 days,1 week,2 weeks,3 weeks,1 month,2 months and 3 months after operation,respectively.Histopathological examination on corneal grafts was performed 1month and 3 months after operation.Results The IOP of all the rabbits lowed after operation,but there was no statistical difference in different time points and various groups (Ftime =29.210,P =0.000; Fgroup =0.254,P =0.938).The grafts of the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group showed varied degrees of corneal opacity and neovascularization 2-3 weeks after operation,and the degree of graft opacity was aggravated obviously in the fourth week,with the RI 8.60±1.52,8.60±0.55 and 8.80±0.84 individually.Neovascularization of the grafts in the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group was found 3 weeks after operation,and the RI were 4.40±0.89,3.20±0.84 and 3.00±0.71,showing a significantly lower than that of the control groups (P＜0.05).Mild inflammatory response of the grafts was seen in the anterior chamber CsA injection group,but it was lightened over time.The histopathological examination revealed obvious thickening of corneal grafts,neovascularization and infiltration of lots of inflammatory cells in the blank control group,subconjunctival vector injection group and the anterior chamber vector injection group.However,only slight new blood vessels and inflammatory response were seen on the subconjunctival CsA injection group,anterior chamber CsA injection group,and 1％ CsA eye drops group.Conclusions Administration of CsA in different methods can prevent the immune rejection after PKP in rabbit eyes,the effect of giving drugs via anterior chamber injection is better than that via subconjunctive pathyway.