鼠眼碱烧伤后羊膜移植对角膜缘干细胞增殖效应的研究
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摘要
目的：制备SD大鼠眼碱烧伤动物模型,行羊膜移植术,观察P63、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)在角膜缘处的表达,进一步了解羊膜对角膜缘干细胞(LSCs)增殖效应的影响。方法：健康SD大鼠40只40眼,实验组标本20只,20眼,行眼碱烧伤加羊膜移植术；对照组标本20只,20眼,单纯行眼碱烧伤；术后第1、2、3、4周分别取角膜缘组织进行固定、石蜡包埋、切片,采用免疫组化技术检测角膜缘组织中P63、PCNA的表达情况。结果：羊膜移植组角膜缘组织中P63、PCNA表达情况显著高于对照组,羊膜移植组和对照组相比较,有统计学意义(P<0.05)；羊膜移植术后1周、2周、3周和4周P63、PCNA蛋白在角膜缘干细胞的表达不一致。呈线性趋势,1周达到高峰,以后逐渐降低。
结论：眼碱烧伤后行羊膜移植术,对角膜缘干细胞有明显的保护作用和并可促进干细胞的增殖表达。
Abstract Objective:To established the animal model of ocular alkali burns in SD rats, and observed the expression of the P63 and proliferating cell nuclear antigen on the corneal limbus.Present study was to discuss the effect of corneal limbal stem cell(LSCs) proliferation by transplantation of amniotic membrane after alkali burn.Methods:A total of 40 healthy SD rats were used for ocular alkali burn models,and divided into two groups randomly.20 rats (eyes) were the experimental group, proceeding amniotic membrane transplantation after ocular alkali burns; 20 rats (eyes) were the control group, simply proceeding ocular alkali burn.The tissue the corneal limbus was used to fixed, paraffin embedded, sections 1、2、3、4 weeks after the operation.The expression of P63 and PCAN in the corneal limbal stem cells were detected by immunohistochemistry. Results:The expression of P63 and PCNA in experimental group was significantly higher than in control group,the difference was statistically significant (P<0.05); the expression of the P63 and PCNA in LSCs decreased gradually along with time of 1,2,3,4 weeks after alkali burn, the peak is 1 week.Conclusion:The amniotic membrane transplantation have significant protective effects to corneal limbal stem cells,and may contribute to the expression of stem cells proliferation after alkali burn.
结论：眼碱烧伤后行羊膜移植术,对角膜缘干细胞有明显的保护作用和并可促进干细胞的增殖表达。
Abstract Objective:To established the animal model of ocular alkali burns in SD rats, and observed the expression of the P63 and proliferating cell nuclear antigen on the corneal limbus.Present study was to discuss the effect of corneal limbal stem cell(LSCs) proliferation by transplantation of amniotic membrane after alkali burn.Methods:A total of 40 healthy SD rats were used for ocular alkali burn models,and divided into two groups randomly.20 rats (eyes) were the experimental group, proceeding amniotic membrane transplantation after ocular alkali burns; 20 rats (eyes) were the control group, simply proceeding ocular alkali burn.The tissue the corneal limbus was used to fixed, paraffin embedded, sections 1、2、3、4 weeks after the operation.The expression of P63 and PCAN in the corneal limbal stem cells were detected by immunohistochemistry. Results:The expression of P63 and PCNA in experimental group was significantly higher than in control group,the difference was statistically significant (P<0.05); the expression of the P63 and PCNA in LSCs decreased gradually along with time of 1,2,3,4 weeks after alkali burn, the peak is 1 week.Conclusion:The amniotic membrane transplantation have significant protective effects to corneal limbal stem cells,and may contribute to the expression of stem cells proliferation after alkali burn.
引文
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22.Zhou Chen,Cintia S,De Paiva,etal.Charaterization of putati-ve stem cell phenotype in human limbal epithelia[J].Stem Cells 2004;22:355-366
23.Elias E,Hemandez G,Carsten T,etal.Correlation of cell cycle kinetics with P63 expressi-on in human limbal epithel-ial cells expanded on intact human amnioitc membrane[J]. Ophthalmic Res 2009;41:83-90
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25.Carmen L,Insausti,Miguel Blanquer,et al.The amniotic membrane as a source of stem cells[J].Histo Histopathol 2010; 25:91-98
26. Gan L, Fagerholm P, Ekenbark S,et al.Expression of proliferating cell nuclear antigen
in comeaskept in long term culture[J]. ActaOphthalmol Scand,1998;76(3):308-313.
27. MaticM, EvansWH, Brink PR, et al.Epidermal stem cells do not communicate thro-ugh gap junctions[J]. J InvestDermatol,2002;118(1):110-116
28.易敬林,钟文贤.不同来源人角膜缘干细胞的体外培养研究[J].江西医学院学报,2005；45(6)：16-31
29.李红,苏敏.PCNA在新西兰大耳兔角膜及角膜缘的表达.局部手术学杂志,2009；18(5)：297-303
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