羊膜对人结膜鳞状上皮化生的影响及其作用机制的实验研究

发布时间：2018-04-03 18:29

本文选题：羊膜　切入点：鳞状上皮化生　出处：《南华大学》2010年硕士论文

【摘要】： 目的:鳞状上皮化生是发生在眼表疾病的一种常见的病理改变,通过体外空气暴露培养人结膜组织,建立结膜鳞状上皮化生模型,探讨羊膜对人结膜鳞状上皮化生的影响及其作用机制。方法:在体外空气暴露条件下培养人正常结膜组织或人正常结膜组织与羊膜或羊膜提取液共培养4天, 8天或12天,应用K10,MUC5AC,Pax6,Ki67,K14,K16和p63检测结膜上皮细胞的分化和增殖状态,同时对Wnt信号通路相关蛋白β-catenin和phosphor-β-catenin进行免疫染色检测。结果:在体外空气暴露培养条件下,结膜鳞状上皮化生被成功诱导:上皮层次明显增加,K10和K16阳性表达, Ki67、K14和p63表达明显上调,MUC5AC和Pax6表达明显下调或消失。与单独培养的结膜相比,与羊膜共培养的结膜,尽管MUC5AC表达下调,但结膜上皮细胞的异常分化和增殖同时被明显抑制。已发生鳞状上皮化生的结膜与羊膜共培养12天后,结膜上皮几乎被逆转成正常的结膜上皮类型。此外羊膜提出液对结膜鳞状上皮化生的发生同样具有明显抑制作用。在体外空气暴露培养条件下,Wnt信号通路在结膜鳞状上皮化生过程中被明显激活,但在与羊膜共培养的结膜上皮中,Wnt激活信号表达明显下调。结论:体外空气暴露培养结膜组织能成功并稳定地诱导出结膜鳞状上皮化生。羊膜能有效抑制和逆转结膜鳞状上皮化生;羊膜组织中可溶性成分-羊膜提取液也能有效抑制结膜鳞状上皮化生;羊膜抑制结膜鳞状上皮化生可能通过羊膜组织中的可溶性成分发挥作用,同时羊膜抑制结膜鳞状上皮化生可能与其抑制Wnt信号通路的活化相关;这将为结膜鳞状上皮化生性疾病的预防和治疗开辟新的途径。
[Abstract]:Objective: squamous metaplasia is a common pathological change in ocular surface diseases. The model of conjunctival squamous metaplasia was established by cultured human conjunctival tissue by air exposure in vitro.To investigate the effect of amniotic membrane on human conjunctival squamous metaplasia and its mechanism.Methods: cultured human normal conjunctival tissue or human normal conjunctival tissue were co-cultured with amniotic membrane or amniotic membrane extract for 4 days, 8 days or 12 days.The Wnt signaling pathway related proteins 尾-catenin and phosphor- 尾-catenin were detected by immunostaining.Results: the conjunctival squamous metaplasia was successfully induced by air exposure in vitro. The positive expression of K10 and K16 was significantly increased at the epithelial level, and the expression of Ki67, K14 and p63 was significantly up-regulated. The expression of MUC5AC and Pax6 was down-regulated or disappeared.Compared with conjunctiva cultured alone and conjunctiva co-cultured with amniotic membrane, the abnormal differentiation and proliferation of conjunctival epithelial cells were significantly inhibited, although the expression of MUC5AC was down-regulated.After 12 days of co-culture of the conjunctiva and amniotic membrane, the conjunctival epithelium was almost reversed into the normal type of conjunctival epithelium.In addition, amniotic membrane fluid also inhibited the development of conjunctival squamous metaplasia.Wnt signaling pathway was significantly activated in conjunctival squamous metaplasia under air exposure in vitro, but down-regulated in conjunctival epithelium co-cultured with amniotic membrane.Conclusion: conjunctival squamous metaplasia can be induced successfully and stably by cultured conjunctival tissue after air exposure in vitro.Amniotic membrane can effectively inhibit and reverse conjunctival squamous metaplasia, and amniotic membrane extract can effectively inhibit conjunctival squamous metaplasia.The inhibition of conjunctival squamous metaplasia by amniotic membrane may play a role through soluble components in amniotic tissue, and the inhibition of amniotic membrane on conjunctival squamous metaplasia may be related to its inhibition of activation of Wnt signaling pathway.This will open a new way for the prevention and treatment of conjunctival squamous metaplasia.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R770.2

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