目的：结合体外细胞实验和在体动物实验研究羊膜上皮细胞(amnion epithelial cell，AEC)混悬液对兔角膜上皮细胞(cornea epithelial cell，CEC)生物学行为的影响及对急性角膜碱烧伤的治疗作用。

方法：细胞实验：实验组将AEC及CEC于Transwell小室中培养，上室为AEC混悬液，下室接种CEC，对照组上室仅为培养基，于不同时间点行CCK-8法检测CEC的增殖活性； 免疫细胞化学法检测PCNA表达； 细胞划痕法观察细胞迁移能力。动物实验：采用改良的兔角膜碱烧伤模型制备方法：将10mm直径环钻轻置于角膜中央，向环钻中央加入NaOH溶液，1min后去环钻迅速冲洗兔眼，随机分三组(空白对照、AEC混悬液滴眼液治疗组、结膜下注射治疗组)，于术后每周行裂隙灯显微镜及荧光素钠染色观察角膜情况。第28d摘除眼球并固定行HE染色，免疫组织化学法检测角膜VEGF、mcp-1的表达。

结果：CCK-8法示AEC混悬液作用后的CEC增殖活性增强(P<0.05)； 免疫细胞化学法显示PCNA阳性表达增强(P<0.05)； 划痕试验示AEC干预后CEC迁移加快。动物实验表明，滴眼液组和结膜下注射组的角膜恢复时间明显缩短，CNV面积均少于对照组(P<0.05)，滴眼液组和结膜下注射组之间差异无统计学意义(P>0.05)； HE染色示治疗组角膜组织炎症细胞浸润更少，组织排列更规则； 免疫组化显示组织中VEGF、mcp-1表达更低(P<0.05)，而滴眼液组和结膜下注射组之间差异无统计学意义(P>0.05)。

结论：AEC混悬液促进CEC增殖和迁移，影响CEC生物学行为变化。经AEC混悬液点眼及结膜下注射治疗兔眼碱烧伤后角膜修复加快，且两种方法抑制炎症反应及CNV形成，有望成为治疗急性角膜碱烧伤的新方法。

METHODS: The rabbit's corneal epithelium were cultured in the lower chamber of transwell, and AEC suspension liquid was dropwised in the upper chamber. There was only culture medium in the upper chamber of the control group. The proliferation of rabbit's corneal epithelium was observed with CCK-8 automated colorimetry and the expression of PCNA was detected by immunocytochemistry. We used the scratch wound assay to detect the migration of corneal epithelial cell(CEC). The in vivo models were established by placing a 10mm diameter corneal trephine in the center of the cornea, within 1mol/L NaOH for 1min. We divided those into three groups: treatment group of AEC suspension liquid eye drop, AEC suspension liquid subconjunctival injection and the control group without any treatment. Using the slit-lamp biomicroscope and fluorescence staining to observe the cornea per week. After 28d we took the eyeballs with the HE staining. The expression of VEGF was detected by immunohistochemistry.

RESULTS: The activity of CEC with AEC treatment was much higher than the control group(P<0.05). The expression of PCNA increased in AEC group(P<0.05). And the migration of CEC in the AEC group was faster than the control one. In vivo, the inflammation of the corneal and the CNV of the AEC group were all significantly reduced compared with the control group(P<0.05). There were less invasive cells and more ordered organization arrangement in ACE group observed by the HE staining. The expression of VEGF and mcp-1 in these two AEC treatment groups all significantly decreased compared with the control group(P<0.05).

CONCLUSION: AEC suspension liquid can promote the proliferation and migration of the rabbit's corneal epithelium. The potential of AEC suspension liquid as a therapy for acute corneal alkali burn.