超低温冷藏对体外培养兔角膜缘上皮细胞活性的影响

目的 分析超低温冷藏兔角膜缘上皮细胞活性变化.设计实验性研究.研究对象超低温冷藏兔角膜缘上皮单细胞悬浮液/兔角膜缘上皮单细胞悬浮液.方法 采用NIH 3T3饲细胞培养体系,将角膜缘上皮制成单细胞悬浮液后-182℃冷藏1周,复苏后行体外培养.以角膜缘上皮单细胞悬浮液为对照组.通过测定细胞贴壁时间、分裂周期、MTT值、3H-胸苷值分析细胞代谢和增殖活性,测定单克隆形成率分析干细胞性,采用AE5免疫组织化学方法鉴定培养上皮细胞,HE染色观察培养角膜上皮细胞形态,PAS染色鉴别结膜杯状细胞.主要指标细胞贴壁时间,分裂周期,MTT值,3H-胸苷值,单克隆形成率及角膜上皮细胞形态等.结果 体外培养超低温冷藏、复苏兔角膜缘上皮细胞的细胞贴壁率、分裂周期、单克隆形成分别为(73.16 13.36)%,(25.42±18.73)小时,(13.17±2.87)%,对照组为(74.89±15.72)%,(20.04±9.70)小时,(15.17±3.25)%.细胞代谢、增殖力也较对照组弱(P>0.05).HE染色示细胞形态、结构无差异,AE5免疫组化染色均为阳性,而PAS染色均为阴性.结论 超低温冷藏可降低体外培养兔角膜缘上皮细胞生物学活性,但其影响有限不足以改变细胞生物学特性和在眼表修复中的应用.(眼科,2008,17:108-112)     

角膜缘上皮细胞体外培养、冻存和复苏的实验研究

目的 建立兔角膜缘上皮细胞体外培养、冻存和复苏的方法。方法 兔角膜缘上皮细胞组织块接种培养。取第三代细胞进行冻存。于冻存后第2周，3、6个月复苏细胞。用MTT法测细胞生长曲线。结果 兔角膜缘上皮细胞体外生长良好。培养细胞AE1单克隆抗体染色阳性，PAS染色阴性。冻存细胞复苏成功。冻存细胞复苏后生长曲线良好。结论 兔角膜缘上皮细胞可以在体外培养、冻存和复苏。     

Baicalein对兔角膜上皮细胞的毒性分析

目的探讨12-LOX选择性抑制剂baicalein的药物毒性,为baicalein的临床应用提供参考。方法体外原代培养兔角膜上皮细胞,加入30μmol/L baicalein继续培养2、4、6d,MTT法测定细胞抑制率。制作兔角膜上皮缺损模型,同浓度baicalein制成滴眼药局部应用,观察其对在体兔角膜上皮增生和移行的影响。结果30μmol/L baicalein对体外原代培养兔角膜上皮细胞作用2d的细胞抑制率为2.6%,作用4d、6d对细胞生长没有明显的抑制作用。缺损闭合实验显示此浓度baicalein对在体兔角膜上皮细胞的增生和移行无任何影响。结论30μmol/L baicalein安全,对兔角膜上皮细胞的生长无影响。     

脱细胞猪角膜基质体外支持角膜上皮和基质细胞的生长

目的:探讨脱细胞猪角膜基质体外能否支持兔角膜细胞的生长。方法:体外培养兔角膜上皮细胞和基质细胞,并接种到制备的脱细胞猪角膜基质上,倒置相差显微镜和组织学观察细胞生长情况。结果:上皮细胞能在脱细胞猪角膜基质上贴附生长,10d时可形成2~3层的复层结构。基质细胞在脱细胞猪角膜基质上贴附生长后可向材料深层迁徙。结论:制备的脱细胞猪角膜基质体外可支持兔角膜上皮细胞和基质细胞的生长。     

培养的兔羊膜上皮细胞构建角膜表层移植片的研究

目的应用培养的兔羊膜上皮细胞（AECs）体外构建复层上皮细胞-角膜基质移植材料,探讨利用AECs重建角膜表层的可行性。方法取妊娠晚期新西兰大白兔（27～28孕周）的羊膜,制成AECs单细胞悬液,用含血清和表皮生长因子（EGF）的DMEM／F12培养液培养、传代,利用免疫组织化学单克隆抗体AE1／AE3、AE5检测培养的AECs中细胞角蛋白ck3／12的表达;将体外培养的2～3代兔AECs种植在新鲜兔角膜基质上,利用气-液界面培养法使之复层化,体外构建复层上皮细胞-角膜基质移植材料,进行光学显微镜和扫描电镜观察,并进行免疫组织化学测定。结果体外培养的兔AECs呈现单克隆抗体AE1／AE3、AE5表达阳性,AECs在新鲜兔角膜基质上能形成形态类似于正常角膜上皮细胞的3～5层复层结构,且复层化后的上皮细胞单克隆抗体AE5表达阳性。结论应用培养的AECs能成功构建类似角膜表层的复层上皮细胞-角膜基质移植材料,AECs可能成为重建角膜表层的一种新的细胞来源。     

荧光激活细胞分离技术在角膜缘干细胞研究中的应用

目的探讨荧光激活细胞分离技术（FACS）在体外培养的兔角膜缘上皮细胞生物学特性研究中的应用。方法收集新西兰白兔的角膜缘组织进行组织块培养。采用FACS分离出体外培养的兔角膜缘上皮细胞中的侧群（SP）细胞和非SP细胞。用2%锥虫蓝染色法对分离出的SP细胞进行克隆形成率（CFE）检测以评估SP细胞的活性,用免疫组织化学染色检测K3/12和ABCG2蛋白在SP细胞和非SP细胞中的表达。结果SP细胞在体外培养的兔角膜缘上皮细胞中占（0.22±0.09）%,分离出的SP细胞的CFE为（5.52±0.45）%,而非SP细胞为（0.78±0.73）%,二者差异有统计学意义（t=2.17,P〈0.01）;大部分SP细胞呈K3/K12抗原阴性,ABCG2免疫标记呈阳性;而非SP细胞呈K3/K12抗原阳性,ABCG2免疫标记呈阴性。结论FACS可以应用于体外培养的兔角膜缘上皮细胞的SP细胞分离,分离出的SP细胞具有较强的增生能力。     

复方中药金芪滴眼剂对兔角膜上皮及基质细胞增殖的影响

目的：观察复方中药金芪滴眼剂对兔角膜上皮细胞及兔角膜基质层细胞增殖的影响。方法：建立体外培养兔角膜上皮细胞，兔角膜基质层细胞系，用MTT法检测不同浓度金芪滴眼剂对细胞增殖活性的影响。结果：金芪滴眼剂对兔角膜上皮细胞有显著的促进增殖作用。且随浓度的增加。增殖率加大，对兔角膜基质层细胞无促增殖作用。高浓度时有抑制作用。结论：复方中药金芪滴眼剂对兔角膜上皮细胞有较好的保护作用。     

体外角膜上皮细胞凋亡的研究

为观察离体角膜上皮细胞经传代培养后细胞死亡的规律及确定其性质。本文应用流式细胞仪 ( FACStarplus)检测凋亡细胞的亚 2倍体 DNA以及细胞凋亡的形态学观察 ,对体外培养的每一代兔角膜缘上皮细胞的凋亡情况进行了观察及分析。结果显示 :角膜缘原代上皮细胞被检测出 7.0 0± 2 .2 3 %的凋亡细胞 ;从第 6代开始 ,角膜上皮细胞凋亡数目显著升高 ( P<0 .0 1) ;之后细胞凋亡数目急剧增加 ,第 8～ 9代后细胞出现集体“自杀”现象。透射电镜观察到这些细胞出现了核浓缩、胞膜发泡、凋亡小体等典型细胞凋亡的形态学改变。结论 :离体角膜缘上皮细胞因外部生长环境的改变 ,在培养到第 6代时出现了细胞凋亡现象。     

压力仿生培养对兔角膜内皮细胞调控的研究

目的 探讨体外压力仿生培养系统下不同梯度压力对角膜内皮细胞形态和功能的调控作用。设计 实验性研究。研究对象 兔角膜内皮细胞。 方法 将体外培养的第一代兔角膜内皮细胞分为五组：A组为无压力常规培养（空白对照）,B组为正常压力仿生培养（15 mmHg）,C组为压力波动组,将压力设为15mmHg、25 mmHg、20 mmHg、10 mmHg,D组30 mmHg压力培养,E组50 mmHg压力培养。细胞均培养24h,免疫法鉴定原代角膜内皮细胞,HE染色和电镜观察细胞形态的改变,流式细胞术检测细胞活性。主要指标 角膜内皮细胞的形态、存活率。 结果 获取的所有细胞证实为角膜内皮细胞表型,无角膜上皮细胞及基质细胞污染。五组细胞分别培养24h后,经HE染色和电镜检测发现正常压力微环境培养的角膜内皮细胞排列紧密,六边形细胞居多,细胞表面微绒毛丰富,细胞核染色质丰富,而高压力培养的角膜内皮细胞活性差,细胞间隙加大。经流式细胞术分析显示,正常压力组、30 mmHg组、压力波动组、50 mmHg组的角膜内皮细胞培养24 h后细胞存活率分别为（98.16±0.45）%、（78.83±1.65）%、（70.2±3.54）%、（41.33±0.25）%（P=0.016）。高压力培养组中随着压力的升高和持续时间延长,细胞活性显著下降。结论 正常压力微环境培养对角膜内皮细胞形态和功能具有正向调节作用,而高压力对角膜内皮细胞具有损伤性,并随时间延长而加重。（眼科,2017,26： 56-60）     

猴,兔角膜细胞成分原代培养的研究

目的 建立从同一角膜上上分离培养角膜上皮,基质和内皮细胞的方法。方法 采用消化法培养猴角膜内皮细胞,上皮细胞和兔角膜上皮细胞,组织块培养法培养角膜基质细胞和兔角膜内皮细胞,结果 猴,兔角膜内皮细胞培养１周后,均能形成细胞单层,猴角膜上皮细胞培养３－４天生长旺盛,但难以形成细胞单层,兔上皮细胞生长旺盛,１周后已达融合状态,细胞呈膜状伸出板层伪足,猴,兔基质细胞易培养,１周后融合成单层细胞,结论 从同     

双氢青蒿素对棘阿米巴抑制作用的实验研究

目的 评价双氢青蒿素对棘阿米巴的体外抑制作用。设计 实验研究。研究对象 临床患者角膜分离的阿米巴虫株。方法 体外培养阿米巴虫株,加入不同稀释浓度（3.2%、1.6%、0.8%、0.4%、0.2%、0.1%、0.05%、0.025%和0.0125%）的双氢青蒿素（DHA）共同孵育,同时以洗必泰（CHG）和聚六甲基双胍（PHMB）做对照药物,通过转种阿米巴培养基和LIVE/DEAD染色观察药物对阿米巴活性的抑制作用。主要指标 光学显微镜观察药物作用后虫体形态变化;LIVE/DEAD染色后,荧光显微镜下阿米巴活性情况,计算最小杀包囊浓度（MCC）和最小杀滋养体浓度（MTAC）值,以及各MTAC和MCC水平下阿米巴虫株的累计分布率,即MTAC50或MCC50和MTAC90或MCC90。结果 药物抑制后的阿米巴包囊干瘪皱缩,LIVE/DEAD法观察,受抑制的包囊呈红色,无活性。DHA组MTAC和MCC值范围均为0.05%~1.6%。MTAC50和MCC50值均为0.4%,MTAC90和MCC90值均为1.6%;CHG组MTAC值范围0.0125%~1.6%,MCC值范围0.0125%~0.8%,MTAC50和 MCC50值分别为0.05%和0.1%,MTAC90和MCC90值均为0.8%。PHMB组MTAC值范围0.00625%~0.4%,MCC值范围0.00625%~0.2%,MTAC50和MCC50值均为0.025%,MTAC90和MCC90值均为0.2%。三组药物对不同阿米巴菌株的MCC和MTAC相比较均无统计学差异（P=0.342、0.459、0.469）。三组MTAC值总体比较DHA组最高,CHG组次之,PHMB组最低（F=3.813,P=0.035)。其中,DHA组高于PHMB组（P=0.011）;DHA组和CHG组以及CHG组和PHMB组比较差异无统计学意义（P=0.105和0.297）。三种药物组的MCC值总体比较差异有统计学意义（F=6.672,P=0.004), DHA组比CHG组高,DHA组比PHMB组高（P=0.017和0.001）; CHG组与PHMB组无差异（P=0.325）。结论 双氢青蒿素在体外对棘阿米巴有抑制作用,其临床治疗阿米巴性角膜炎的作用仍待验证。     

Cysticidal effect on acanthamoeba and toxicity on human keratocytes by polyhexamethylene biguanide and chlorhexidine

PURPOSE: To evaluate the cysticidal effect of polyhexamethylene biguanide (PHMB) and chlorhexidine on Acanthamoeba and its toxic effect on cultured human keratocytes. METHODS: Each well of a twofold-diluted Acanthamoeba cyst-containing suspension of 5 x 10(4) cysts/mL was treated with PHMB and chlorhexidine for 8, 24, and 48 hours to determine the minimal cysticidal concentration (MCC) of each disinfectant. Human corneal keratocytes (5 x 10(4) cells/mL) were exposed to PHMB and chlorhexidine for the same time to determine the survival rate of keratocytes. Inverted phase-contrast and electron microscopy were used to observe the morphologic changes. RESULTS: The mean MCC of PHMB for 8, 24, and 48 hours was 9.42, 5.62, and 2.37 microg/mL, respectively. The mean MCC of chlorhexidine for 8, 24, and 48 hours was 24.32, 10.02, and 7.02 microg/mL, respectively. The respective survival rate of keratocytes at the MCC was 91.7%, 64.6%, and 49.7% for PHMB and 95.7%, 90.6%, and 78.1% for chlorhexidine, respectively. The cysts and keratocytes showed more damaged appearances after treatment with PHMB than chlorhexidine. CONCLUSIONS: PHMB and chlorhexidine showed a similar amoebicidal efficacy. However, PHMB seemed to be more toxic to keratocytes than chlorhexidine.     

Comparison of polyhexamethylene biguanide and chlorhexidine as monotherapy agents in the treatment of Acanthamoeba keratitis

PURPOSE: To compare the therapeutic outcomes of polyhexamethylene biguanide (PHMB) and chlorhexidine for Acanthamoeba keratitis. DESIGN: Prospective, double-masked, randomized comparative study. METHODS: Fifty-six eyes of 55 patients with Acanthamoeba keratitis were randomized to receiving PHMB 0.02% or chlorhexidine 0.02%. Diagnosis was made based on positive culture results (cornea or contact lens case) or on clinical grounds. The primary outcome measure was treatment failure defined as failure to induce a favorable clinical response within two weeks. Secondary outcomes were: 1) recovery of visual acuity (VA), 2) the degree of corneal scarring posttreatment, or 3) the need for penetrating keratoplasty. RESULTS: Fifty-one eyes completed the study. Twenty-three eyes received PHMB and 28 received chlorhexidine. Ninety-eight percent were contact lens wearers. Eighteen (78%) PHMB patients were treatment successes compared with 24 (85.7%) chlorhexidine patients (P = .71). Diagnosis was confirmed by positive corneal culture results in 26 cases (51%). Diagnosis was made within 28 days in 29 cases (56.9%), between one and two months in 13 cases (25.5%), and after more than two months in eight cases (15.7%). Improvement in VA was seen in 13 eyes (56.5%) receiving PHMB vs 20 eyes (71.4%) receiving chlorhexidine. Mild one-quadrant corneal scarring was seen in 43.5% of eyes receiving PHMB and 71.4% of eyes receiving chlorhexidine, whereas moderate corneal scarring in two or three quadrants was seen in 21.7% of eyes receiving PHMB and in 10.7% of eyes receiving chlorhexidine. Five eyes worsened while receiving PHMB vs four eyes worsening while receiving chlorhexidine. Penetrating keratoplasty was required in three eyes from PHMB group and in two eyes from chlorhexidine group. CONCLUSIONS: Outcomes were similar when using PHMB and chlorhexidine as monotherapy agents in treating Acanthamoeba keratitis.     

The effect of substance P with insulin-like growth factor-1 on corneal epithelial cell proliferation]

PURPOSE: To elucidate the pathogenesis of neuroparalytic keratopathy, we examined the effect of substance P (SP) on corneal epithelial cell proliferation in comparison with that of insulin-like growth factor-1 (IGF-1) in vivo and in vitro. METHODS: In the in vivo study, the epithelium of rabbit cornea was removed mechanically and treated with eye drops containing SP, IGF-1, SP + IGF-1, or physiological saline as a control. Corneas were excised 48 hours after the removal of the epithelium, labeled with 3H-thymidine, and subjected to autoradiography. An in vitro study was also performed by culturing rabbit corneal epithelial cells in culture medium containing SP with or without IGF-1 and labeling cells with 3H-thymidine for a subsequent autoradiographical study. FINDINGS: In the in vivo study, SP in combination with IGF-1 enhanced the healing of rabbit corneal epithelial defect. 3H-thymidine autoradiography revealed that in both in vivo and in vitro studies, SP + IGF-1 stimulated corneal epithelial cell proliferation, but SP or IGF-1 alone did not. CONCLUSION: These results indicate that SP enhances the stimulatory effect of IGF-1 on corneal epithelial cell proliferation and accelerates corneal epithelial healing.     

以异种角膜脱细胞基质为载体构建角膜上皮-载体-内皮复合物的实验研究

目的探讨以异种角膜脱细胞基质为载体,体外构建生物角膜的可能性和方法。方法应用去垢剂1％TritonX-100冷冻干燥处理制备猪角膜脱细胞基质载体,在其上皮面和内皮面分别接种兔角膜上皮细胞和内皮细胞,体外培养2周。将复合物制成组织切片,在光镜下观察组织形态（HE染色）,采用免疫组织化学方法检测角膜上皮特异性细胞角蛋白3（CK3）,使用锥虫蓝联合茜素红染色观察内皮细胞,在扫描电镜下观察上皮面和内皮面的超微结构。结果体外培养生物角膜获得上皮、无细胞基质和内皮三层复合结构。4或5层复层上皮中以扁平细胞为主,胞质内特异性CK3表达阳性;内皮层为一连续的单层扁平细胞,细胞活性良好,锥虫蓝联合茜素红染色显示组织呈典型的蜂巢样镶嵌结构。扫描电镜下观察,在载体的上皮面细胞呈复层样生长,形态近似扁平与梭形之间;内皮面为多边形单层细胞,表面具有微绒毛结构。结论构建的生物角膜为上皮一脱细胞基质载体一内皮复合物,初具角膜的雏形结构。异种角膜脱细胞基质提供了良好的细胞生长界面,有望成为体外构建角膜的载体材料。     

兔角膜内皮細胞體外原代培養及形熊學觀察

目的研究培養兔角膜内皮細胞,為實驗提供基礎.方法通過改良的細胞收獲、培養技術,在體外進行兔角膜内皮細胞的培養,細胞的類型由電鏡證實.觀察hEGF不同時間點對角膜内皮細胞生長狀態的影響.結果未加入任何促細胞分裂劑的情况下,細胞在體外生長良好,約一周左右形成密集狀態,形態類似天然細胞,經倒置顯微鏡和電鏡證實為角膜内皮細胞.在有hEGF存在時,細胞生長迅速,3-4天形成單層,各個時間點EGF組的細胞數高于對照組.結論該技術可為角膜内皮細胞的研究提供較多的純内皮細胞.     

猪脱细胞角膜基质组织结构特点与生物相容性研究

背景构建组织工程化角膜时,载体的选择十分重要。目前有多种载体可供选用,但脱细胞角膜基质是公认的较好载体。目的观察猪脱细胞角膜基质的组织结构特点,评价其与异种角膜基质和上皮细胞的生物相容性。方法取猪角膜组织进行组织块培养,经胰蛋白酶-EDTA酶消化角膜上皮、基质、内皮细胞,支架组织于-20℃冷冻干燥后灭菌保存。猪脱细胞角膜基质石蜡切片行常规苏木精一伊红染色,光学显微镜下观察组织的形态学特征;扫描电镜下观察其组织结构特点;并对其物理特性,如抗拉性、膨胀性、含水量和透明度进行评价。将猪脱细胞角膜基质移植到兔角膜基质层内,同时与体外培养的兔角膜上皮细胞共培养4周,分析其组织和细胞生物相容性。结果经酶消化处理后猪角膜组织中未见上皮、基质和内皮细胞,其胶原纤维直径大小、排列与正常角膜组织相同,抗拉性、膨胀性、含水量和透明度等物理特性与正常猪角膜相似。猪脱细胞角膜基质行异种兔角膜基质层间移植1周时可见轻度水肿,2周后水肿基本消退,4周时透明度较好。猪脱细胞角膜基质与兔角膜基质之间贴附良好,未见炎症反应及新生血管。兔角膜上皮细胞接种于猪脱细胞角膜基质上共培养4周后CK3表达阳性。猪脱细胞角膜基质脱水前与脱水2h、4h后及正常猪角膜基质间的抗拉性、膨胀性、含水量的差异均无统计学意义（P〉0．05）,但脱水2h、4h后及正常猪角膜基质的透明度明显好于脱水前,差异均有统计学意义（P〈0．01）。结论猪脱细胞角膜基质组织结构与正常猪角膜相似,与兔角膜基质和上皮细胞具有良好的生物相容性。     

Epithelial-stromal interactions: specific stimulation of corneal epithelial cell growth in vitro by a factor(s) from cultured stromal fibroblasts

By using a recently modified method of isolating and culturing rabbit corneal cells, this study investigated the presence of a diffusible substance(s) in stromal fibroblast conditioned medium that stimulated the growth of cultured corneal epithelial cells. The growth stimulation involved initiation of DNA synthesis (assayed by [3H]-thymidine incorporation) and enhanced cell proliferation (quantified by cell counting). Among the three corneal cell types, only fibroblasts (rabbit and human) released the stimulatory substance, which acted only on epithelial cells. The effect of this stromal fibroblast factor (SFF) was observed after an exposure period of less than 16 hr and persisted as long as it was present. Its action was concentration-dependent and was not a result of improvement in the survival of epithelial cells during culture. Both sparse and confluent epithelial cultures were susceptible to SFF. The release of SFF was correlated with the number of fibroblasts in the culture and appeared to be sensitive to the growth condition of the cells. Both the release and action of SFF did not depend on the presence of serum in the culture medium. The factor was heat resistant and insensitive to proteolytic enzymes. From ultrafiltration studies, the size of SFF was estimated to be in the approximate range of 50-1000 daltons. By direct comparison of the stimulatory effect with other previously studied growth promoting agents, it was concluded that SFF was not epidermal growth factor, fibroblast growth factor, putrescine, cyclic AMP, hydrocortisone or acetate. The implication of SFF in the regulation of epithelial growth by endogenous, intercellular mechanisms is discussed.     

Spherical indentations of human and rabbit corneal epithelium following extended contact lens wear.

PURPOSE: Mucin balls appear to cause spherical indentations in the corneal epithelium during silicone hydrogel extended contact lens wear. The purpose of this report is to describe and quantify these spherical indentations, as examined in the human cornea by in vivo confocal microscopy and by in vitro immunocytochemistry in the rabbit cornea. METHODS: Confocal images of full-thickness corneal epithelium were taken from three human patients participating in a 1-year extended contact lens-wear trial. Diameter and depth of the indentations were determined and measured. Two rabbit corneas showing identical indentations were stained with propidium iodide (nuclear stain) and Ki-67 (proliferation marker) and were examined using a laser scanning confocal microscope. RESULTS: The diameter of the spherical indentations is largest on the epithelial surface, ranging from 33.9 to 78.8 microm. Indentations form spherical sections whose depth variably extends into the corneal epithelium, reaching as far as the basal lamina. The rabbit model showed no epithelial nuclei within the indentation. Furthermore, stromal cells localized immediately beneath the indentations were positive for Ki-67 (proliferation). DISCUSSION: Spherical indentations of the corneal epithelium induced by mucin balls appear to be gaps or holes that can extend deep into the corneal epithelium. Indentations may potentially open a pathway for infectious microorganisms to penetrate the cornea. Surprisingly, stromal cells immediately beneath the holes were stimulated to proliferate, and there seemed to be an increase in localized cell density.     

内皮素-1对培养的兔角膜内皮细胞的影响

目的 研究内皮素-1(ET-1)对培养的兔角膜内皮细胞的增殖能力和移行能力的影响。方法 采用MTT方法并通过损伤模型观察不同浓度ET-1加药48 h后，对体外培养的兔角膜内皮细胞增殖和细胞移行的能力。光镜下和透射电镜下观察ET-1对兔角膜内皮细胞形态的影响。采用免疫组化染色法和计算机图像分析系统检测不同浓度ET-1对细胞PCNA表达的影响。结果 一定浓度ET-1促进培养的兔角膜内皮细胞的增殖和移行，且呈剂量相关性。10 pmol／L时起作用，200 pmol／L时发挥最大作用。光镜下和透射电镜下发现ET-1对兔角膜内皮细胞的形态学特征和超微结构无影响。兔角膜内皮细胞PCNA表达阳性，ET-1促进其表达，且呈剂量相关性。结论 ET-1可作为一种生长因子，一定浓度下可促进培养的兔角膜内皮细胞增殖和移行能力。