發(fā)布時(shí)間：2018-06-03 08:26

  本文選題：犬 + 角膜損傷��； 參考：《揚(yáng)州大學(xué)》2015年碩士論文

【摘要】：犬角膜損傷是臨床常見(jiàn)的眼科疾病,且致盲率較高。羊膜免疫源性低,以其優(yōu)越的理化特性而成為理想的生物移植材料。因此,為了探索犬角膜損傷的高效療法,本試驗(yàn)首先建立了犬角膜堿燒傷疾病模型,然后采用羊膜移植術(shù)對(duì)其進(jìn)行治療,并觀察病犬眼睛的臨床癥狀、角膜組織學(xué)變化及相關(guān)細(xì)胞因子TNF-α、VEGF和TGF-β1的基因相對(duì)表達(dá)量情況。選取健康成年比格犬6只,將直徑6 mm的圓形濾紙片用1 mol/L NaOH溶液浸濕,置于角膜中央1 min,然后立即用生理鹽水沖洗眼球表面,制得犬角膜堿燒傷疾病模型。取剖腹產(chǎn)犬胎兒羊膜,制備犬羊膜片,置于DMEM甘油液中,-80℃保存。本試驗(yàn)用同一條犬的左眼作為對(duì)照組,僅進(jìn)行堿燒傷；右眼為試驗(yàn)組,堿燒傷后進(jìn)行羊膜移植。術(shù)后每日觀察犬角膜水腫、角膜新生血管和眼分泌物等情況,于術(shù)后3、7、14、21、28和35 d測(cè)量眼內(nèi)壓和淚液量,并對(duì)角膜混濁、新生血管及上皮修復(fù)情況進(jìn)行評(píng)分；術(shù)后35 d采集犬角膜組織,將其分成2份,一份用于病理組織學(xué)觀察,組織切片用HE染色及Masson三色染色；另一份用于熒光定量PCR實(shí)驗(yàn),檢測(cè)TNF-α、VEGF和TGF-β1的基因相對(duì)表達(dá)量。試驗(yàn)結(jié)果如下：(1)術(shù)后兩組犬的角膜都出現(xiàn)不同程度的水腫和眼分泌物增多癥狀,試驗(yàn)組犬眼球表面的羊膜植片較平整,羊膜于1周內(nèi)溶解,術(shù)后5 d角膜緣出現(xiàn)新生血管芽,至第16 d時(shí)眼分泌物減少并恢復(fù)到正常水平；而對(duì)照組犬眼術(shù)后5 d角膜緣出現(xiàn)刷狀的新生血管,至第28 d時(shí)仍有2只眼睛有較多的分泌物,在第6 d時(shí)有一只眼的角膜出現(xiàn)穿孔。至術(shù)后35 d時(shí),試驗(yàn)組犬眼在角膜混濁程度及血管新生方面明顯優(yōu)于對(duì)照組(P0.05),第21 d時(shí)兩組犬角膜熒光素染色出現(xiàn)顯著差異(P0.05),試驗(yàn)組犬角膜著色明顯輕于對(duì)照組,至術(shù)后35 d時(shí)試驗(yàn)組犬角膜熒光素著色不明顯,對(duì)照組犬角膜呈淺著色。術(shù)后14 d,試驗(yàn)組犬眼的眼內(nèi)壓及淚液量指標(biāo)升高,但處于正常生理范圍之內(nèi)；對(duì)照組犬眼的眼內(nèi)壓極顯著降低(P0.01),淚液量顯著減少(P0.05)；至第35 d時(shí)兩組犬眼的眼內(nèi)壓及淚液量均恢復(fù)至正常水平,但對(duì)照組處于正常生理范圍的下限。(2)術(shù)后35 d,犬角膜病理組織學(xué)切片顯示：對(duì)照組犬角膜有較多的炎性細(xì)胞和新生血管,血管腔內(nèi)有成熟紅細(xì)胞,成纖維細(xì)胞肥大,基底細(xì)胞腫脹且間隙增大,膠原纖維排列紊亂；試驗(yàn)組犬角膜炎性細(xì)胞和新生血管相對(duì)較少,成纖維細(xì)胞扁平,膠原纖維排列相對(duì)較整齊。(3)術(shù)后35 d,試驗(yàn)組犬角膜組織中TNF-α、VEGF和TGF-β1的基因相對(duì)表達(dá)量極顯著低于對(duì)照組(P0.01)。由上述試驗(yàn)結(jié)果可知,羊膜具有抑制炎癥反應(yīng)、抑制血管新生和瘢痕的形成、促進(jìn)角膜損傷愈合的重要作用,對(duì)犬角膜堿燒傷有一定的臨床療效。
[Abstract]:Canine corneal injury is a common ophthalmic disease and has a high rate of blindness. Amniotic membrane with low immunogenicity is an ideal biological transplantation material because of its superior physical and chemical properties. Therefore, in order to explore the effective treatment of canine corneal injury, we first established the canine corneal alkali burn disease model, then treated it with amniotic membrane transplantation, and observed the clinical symptoms of the canine eyes. The changes of corneal histology and the relative expression of VEGF and TGF- 尾 1 genes in the related cytokines TNF- 偽 and TGF- 尾 1. Six healthy adult Beagle dogs were selected. The circular filter paper with diameter of 6 mm was soaked in 1 mol/L NaOH solution and placed in the center of cornea for 1 minute. The corneal alkali burn model was made by rinsing the surface of the eyeball with physiological saline immediately. The amniotic membrane of caesarean section was prepared and stored at 80 鈩，

本文編號(hào)：1972131