背景 角膜上皮损伤可导致角膜溃疡和基质混浊,甚至出现不可逆的视功能损害,以往采用的药物治疗仅能缓解刺激症状,而促进角膜上皮细胞再生的作用较弱,因此寻求能有效调控角膜上皮细胞生长,从而有效治疗角膜上皮损伤的药物至关重要. 目的 探讨重组人BIGH3蛋白滴眼液对角膜上皮损伤的修复作用.方法 50只清洁级健康成年新西兰大白兔,均取右眼为实验眼,其中2只兔制作角膜上皮损伤模型后即刻行裂隙灯显微镜检查及角膜荧光素染色,眼前节照相后处死；48只兔按照随机数字表法随机分为重组人表皮生长因子(EGF)衍生物组(阳性对照组)、生理盐水组(阴性对照组)、质量分数0.25％重组人BIGH3蛋白滴眼液组及0.5％重组人BIGH3蛋白滴眼液组,每组12只.在直径7 mm环钻内滴入体积分数20％乙醇,置于角膜中央区60 s,然后用角膜刮匙刮去角膜上皮,制作角膜上皮缺损模型.术后各组兔眼分别点用相应药物,每天4次,分别于术后12、24、36、48、72 h行裂隙灯显微镜检查及角膜荧光素染色,于12、24、36、48 h采用抽签法各处死2只实验兔,72 h时各处死4只实验兔,制备角膜组织标本并行苏木精-伊红染色,观察并比较各组兔眼在不同时间点角膜上皮修复情况,应用计算机图像处理系统测量各组兔眼不同时间点角膜上皮愈合面积.结果 各组兔眼用药后均未发现任何眼部刺激症状.组织病理学研究发现,造模后即刻全层角膜上皮缺损,随着时间的延长,实验组及对照组兔角膜上皮缺损区皆逐渐变小,可见周边角膜上皮向中央缺损区移行,细胞层数逐渐增加,细胞排列极向逐渐趋于规则.重组人EGF衍生物组、生理盐水组、0.25％及0.5％重组人BIGH3蛋白滴眼液组角膜上皮愈合速率分别为15.00、13.81、18.05、18.86 mm2/d.与生理盐水组比较,术后12、24、36、48 h,0.25％、0.5％重组人BIGH3蛋白滴眼液组及重组人EGF衍生物组角膜损伤面积明显缩小,差异均有统计学意义(均P=0.000),但造模12、24、36 h,重组人EGF衍生物组与生理盐水组比较差异均无统计学意义(P=0.321、0.057、0.126).与重组人EGF衍生物组比较,术后各时间点0.5％重组人BIGH3蛋白滴眼液组角膜损伤面积明显缩小,差异均有统计学意义(P=0.042、0.039、0.025、0.008),0.25％重组人BIGH3蛋白滴眼液组角膜损伤面积仅在术后12h、24 h明显缩小,差异均有统计学意义(P=0.047、0.042).术后各时间点0.25％、0.5％重组人BIGH3蛋白滴眼液组间角膜损伤面积的差异均无统计学意义(P=0.358、0.259、0.108、0.062).术后72 h,生理盐水组角膜损伤面积为(0.51±0.42)mm2,而其他3个组角膜上皮均完全修复.角膜上皮修复曲线表明,0.25％及0.5％重组人BIGH3蛋白滴眼液组角膜上皮修复的相对面积均优于重组人EGF衍生物组和生理盐水组. 结论 0.25％及0.5％重组人BIGH3蛋白滴眼液对角膜上皮损伤的愈合有明显的促进作用,其作用优于EGF.
揹景 角膜上皮損傷可導緻角膜潰瘍和基質混濁,甚至齣現不可逆的視功能損害,以往採用的藥物治療僅能緩解刺激癥狀,而促進角膜上皮細胞再生的作用較弱,因此尋求能有效調控角膜上皮細胞生長,從而有效治療角膜上皮損傷的藥物至關重要. 目的 探討重組人BIGH3蛋白滴眼液對角膜上皮損傷的脩複作用.方法 50隻清潔級健康成年新西蘭大白兔,均取右眼為實驗眼,其中2隻兔製作角膜上皮損傷模型後即刻行裂隙燈顯微鏡檢查及角膜熒光素染色,眼前節照相後處死；48隻兔按照隨機數字錶法隨機分為重組人錶皮生長因子(EGF)衍生物組(暘性對照組)、生理鹽水組(陰性對照組)、質量分數0.25％重組人BIGH3蛋白滴眼液組及0.5％重組人BIGH3蛋白滴眼液組,每組12隻.在直徑7 mm環鑽內滴入體積分數20％乙醇,置于角膜中央區60 s,然後用角膜颳匙颳去角膜上皮,製作角膜上皮缺損模型.術後各組兔眼分彆點用相應藥物,每天4次,分彆于術後12、24、36、48、72 h行裂隙燈顯微鏡檢查及角膜熒光素染色,于12、24、36、48 h採用抽籤法各處死2隻實驗兔,72 h時各處死4隻實驗兔,製備角膜組織標本併行囌木精-伊紅染色,觀察併比較各組兔眼在不同時間點角膜上皮脩複情況,應用計算機圖像處理繫統測量各組兔眼不同時間點角膜上皮愈閤麵積.結果 各組兔眼用藥後均未髮現任何眼部刺激癥狀.組織病理學研究髮現,造模後即刻全層角膜上皮缺損,隨著時間的延長,實驗組及對照組兔角膜上皮缺損區皆逐漸變小,可見週邊角膜上皮嚮中央缺損區移行,細胞層數逐漸增加,細胞排列極嚮逐漸趨于規則.重組人EGF衍生物組、生理鹽水組、0.25％及0.5％重組人BIGH3蛋白滴眼液組角膜上皮愈閤速率分彆為15.00、13.81、18.05、18.86 mm2/d.與生理鹽水組比較,術後12、24、36、48 h,0.25％、0.5％重組人BIGH3蛋白滴眼液組及重組人EGF衍生物組角膜損傷麵積明顯縮小,差異均有統計學意義(均P=0.000),但造模12、24、36 h,重組人EGF衍生物組與生理鹽水組比較差異均無統計學意義(P=0.321、0.057、0.126).與重組人EGF衍生物組比較,術後各時間點0.5％重組人BIGH3蛋白滴眼液組角膜損傷麵積明顯縮小,差異均有統計學意義(P=0.042、0.039、0.025、0.008),0.25％重組人BIGH3蛋白滴眼液組角膜損傷麵積僅在術後12h、24 h明顯縮小,差異均有統計學意義(P=0.047、0.042).術後各時間點0.25％、0.5％重組人BIGH3蛋白滴眼液組間角膜損傷麵積的差異均無統計學意義(P=0.358、0.259、0.108、0.062).術後72 h,生理鹽水組角膜損傷麵積為(0.51±0.42)mm2,而其他3箇組角膜上皮均完全脩複.角膜上皮脩複麯線錶明,0.25％及0.5％重組人BIGH3蛋白滴眼液組角膜上皮脩複的相對麵積均優于重組人EGF衍生物組和生理鹽水組. 結論 0.25％及0.5％重組人BIGH3蛋白滴眼液對角膜上皮損傷的愈閤有明顯的促進作用,其作用優于EGF.
배경 각막상피손상가도치각막궤양화기질혼탁,심지출현불가역적시공능손해,이왕채용적약물치료부능완해자격증상,이촉진각막상피세포재생적작용교약,인차심구능유효조공각막상피세포생장,종이유효치료각막상피손상적약물지관중요. 목적 탐토중조인BIGH3단백적안액대각막상피손상적수복작용.방법 50지청길급건강성년신서란대백토,균취우안위실험안,기중2지토제작각막상피손상모형후즉각행렬극등현미경검사급각막형광소염색,안전절조상후처사；48지토안조수궤수자표법수궤분위중조인표피생장인자(EGF)연생물조(양성대조조)、생리염수조(음성대조조)、질량분수0.25％중조인BIGH3단백적안액조급0.5％중조인BIGH3단백적안액조,매조12지.재직경7 mm배찬내적입체적분수20％을순,치우각막중앙구60 s,연후용각막괄시괄거각막상피,제작각막상피결손모형.술후각조토안분별점용상응약물,매천4차,분별우술후12、24、36、48、72 h행렬극등현미경검사급각막형광소염색,우12、24、36、48 h채용추첨법각처사2지실험토,72 h시각처사4지실험토,제비각막조직표본병행소목정-이홍염색,관찰병비교각조토안재불동시간점각막상피수복정황,응용계산궤도상처리계통측량각조토안불동시간점각막상피유합면적.결과 각조토안용약후균미발현임하안부자격증상.조직병이학연구발현,조모후즉각전층각막상피결손,수착시간적연장,실험조급대조조토각막상피결손구개축점변소,가견주변각막상피향중앙결손구이행,세포층수축점증가,세포배렬겁향축점추우규칙.중조인EGF연생물조、생리염수조、0.25％급0.5％중조인BIGH3단백적안액조각막상피유합속솔분별위15.00、13.81、18.05、18.86 mm2/d.여생리염수조비교,술후12、24、36、48 h,0.25％、0.5％중조인BIGH3단백적안액조급중조인EGF연생물조각막손상면적명현축소,차이균유통계학의의(균P=0.000),단조모12、24、36 h,중조인EGF연생물조여생리염수조비교차이균무통계학의의(P=0.321、0.057、0.126).여중조인EGF연생물조비교,술후각시간점0.5％중조인BIGH3단백적안액조각막손상면적명현축소,차이균유통계학의의(P=0.042、0.039、0.025、0.008),0.25％중조인BIGH3단백적안액조각막손상면적부재술후12h、24 h명현축소,차이균유통계학의의(P=0.047、0.042).술후각시간점0.25％、0.5％중조인BIGH3단백적안액조간각막손상면적적차이균무통계학의의(P=0.358、0.259、0.108、0.062).술후72 h,생리염수조각막손상면적위(0.51±0.42)mm2,이기타3개조각막상피균완전수복.각막상피수복곡선표명,0.25％급0.5％중조인BIGH3단백적안액조각막상피수복적상대면적균우우중조인EGF연생물조화생리염수조. 결론 0.25％급0.5％중조인BIGH3단백적안액대각막상피손상적유합유명현적촉진작용,기작용우우EGF.
Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25％ or 0.5％ recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25％ and 0.5％ recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25％,0.5％ recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5％recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25％ recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25％ and 0.5％recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25％ and 0.5％ recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.
