本文選題：羊膜　切入點(diǎn)：鱗狀上皮化生　出處：《南華大學(xué)》2010年碩士論文

【摘要】： 目的:鱗狀上皮化生是發(fā)生在眼表疾病的一種常見(jiàn)的病理改變,通過(guò)體外空氣暴露培養(yǎng)人結(jié)膜組織,建立結(jié)膜鱗狀上皮化生模型,探討羊膜對(duì)人結(jié)膜鱗狀上皮化生的影響及其作用機(jī)制。 方法:在體外空氣暴露條件下培養(yǎng)人正常結(jié)膜組織或人正常結(jié)膜組織與羊膜或羊膜提取液共培養(yǎng)4天, 8天或12天,應(yīng)用K10,MUC5AC,Pax6,Ki67,K14,K16和p63檢測(cè)結(jié)膜上皮細(xì)胞的分化和增殖狀態(tài),同時(shí)對(duì)Wnt信號(hào)通路相關(guān)蛋白β-catenin和phosphor-β-catenin進(jìn)行免疫染色檢測(cè)。 結(jié)果:在體外空氣暴露培養(yǎng)條件下,結(jié)膜鱗狀上皮化生被成功誘導(dǎo):上皮層次明顯增加,K10和K16陽(yáng)性表達(dá), Ki67、K14和p63表達(dá)明顯上調(diào),MUC5AC和Pax6表達(dá)明顯下調(diào)或消失。與單獨(dú)培養(yǎng)的結(jié)膜相比,與羊膜共培養(yǎng)的結(jié)膜,盡管MUC5AC表達(dá)下調(diào),但結(jié)膜上皮細(xì)胞的異常分化和增殖同時(shí)被明顯抑制。已發(fā)生鱗狀上皮化生的結(jié)膜與羊膜共培養(yǎng)12天后,結(jié)膜上皮幾乎被逆轉(zhuǎn)成正常的結(jié)膜上皮類(lèi)型。此外羊膜提出液對(duì)結(jié)膜鱗狀上皮化生的發(fā)生同樣具有明顯抑制作用。在體外空氣暴露培養(yǎng)條件下,Wnt信號(hào)通路在結(jié)膜鱗狀上皮化生過(guò)程中被明顯激活,但在與羊膜共培養(yǎng)的結(jié)膜上皮中,Wnt激活信號(hào)表達(dá)明顯下調(diào)。 結(jié)論:體外空氣暴露培養(yǎng)結(jié)膜組織能成功并穩(wěn)定地誘導(dǎo)出結(jié)膜鱗狀上皮化生。羊膜能有效抑制和逆轉(zhuǎn)結(jié)膜鱗狀上皮化生;羊膜組織中可溶性成分-羊膜提取液也能有效抑制結(jié)膜鱗狀上皮化生;羊膜抑制結(jié)膜鱗狀上皮化生可能通過(guò)羊膜組織中的可溶性成分發(fā)揮作用,同時(shí)羊膜抑制結(jié)膜鱗狀上皮化生可能與其抑制Wnt信號(hào)通路的活化相關(guān);這將為結(jié)膜鱗狀上皮化生性疾病的預(yù)防和治療開(kāi)辟新的途徑。
[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.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R770.2

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本文編號(hào)：1706401