本文選題：角膜　切入點：堿燒傷　出處：《南昌大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

【摘要】： 目的:研究他可莫司(FK506)滴眼液治療大鼠角膜堿燒傷的藥物有效濃度及給藥方案,評價其抑制角膜堿燒傷后免疫炎癥反應(yīng),改善角膜微環(huán)境的效能,并探討其作用機制。 方法:NaOH濾紙片敷貼法制作大鼠角膜中度堿燒傷模型,96只wistar大鼠隨機分為A、B、C、D四組,A、B、C組為治療組,給予濃度0.05%、0.1%、0.5%FK506滴眼液治療,4次/天,D組為陰性對照組。觀察不同組別大鼠角膜透明性、基質(zhì)融解程度以及新生血管形成情況。在不同時間點(第3,7,14,30d)處死大鼠取下角膜, HE染色分析角膜組織結(jié)構(gòu)的變化,比較炎癥細胞浸潤程度的差異,免疫熒光法檢測CD4/CD8陽性T細胞時空表達規(guī)律。 結(jié)果:堿液濾紙片燒傷角膜后,角膜霧狀水腫,燒傷區(qū)角膜上皮缺失,基質(zhì)融解形成潰瘍,角膜緣血管擴張,后期形成新生血管,角膜失去透明性。FK506治療組的角膜在潰瘍形成面積,角膜混濁等級以角膜新生血管化程度方面均明顯好于對照組。①.角膜融解定量分析結(jié)果:燒傷后3d,A、B、C組角膜融解面積分別為(3.14±0.25)mm2、(2.65±0.28) mm2、(2.27±0.47) mm2,而對照組D組為(4.06±0.41) mm2,差異有統(tǒng)計學(xué)意義(P0.05),低濃度FK506治療組與中、高濃度組相比亦存在統(tǒng)計學(xué)差異(P0.05),而0.1%與0.5%濃度組之間則無明顯差別(P0.05); 7d時角膜融解面積縮小,分別為A組(1.82±0.51)mm2、B組(1.34±0.41) mm2、C組(0.90±0.39) mm2,與對照組D組(2.47±0.54) mm2,相比差異有統(tǒng)計學(xué)意義(P0.05)。②.角膜混濁定量分析結(jié)果:治療組在燒傷后角膜混濁評分低于對照組(P0.05),其規(guī)律與角膜融解面積的變化曲線相一致,中高濃度組之間則無差異。③.角膜新生血管定量分析結(jié)果:治療組在燒傷后3d角膜新生血管面積分別為A組(4.26±0.36)mm2、B組(3.88±0.33) mm2、C組(3.54±0.46) mm2均小于對照組D組(5.00±0.12) mm2,差異有顯著意義(P0.01),A與B、C比較也有統(tǒng)計學(xué)意義(P0.05),而B與C比較無統(tǒng)計學(xué)意義(P0.05);此種差異性保持至第14d方逐漸減小。燒傷角膜HE染色檢查顯示FK506治療組基質(zhì)中炎癥細胞浸潤的數(shù)量明顯少于對照組,計數(shù)結(jié)果顯示3d時,A組(77.67±4.51)個/0.04mm2、B組(51.33±3.21)個/0.04mm2、C組(41.67±3.06)個/0.04mm2,而同期對照組D組(97.00±6.25)個/0.04mm2,差異均有統(tǒng)計學(xué)意義(P0.05)。免疫熒光提示治療組中CD4陽性細胞的數(shù)量少于對照組,通過光密度分析,低濃度治療組為中等強度熒光,中高濃度治療組為低熒光,而對照組則表現(xiàn)為彌漫性強熒光。在整個角膜堿燒傷病理過程中,CD8+ T淋巴細胞的表達情況,給藥組與對照組對比未見明顯差異。 結(jié)論:他可莫司(FK506)局部使用可明顯下調(diào)角膜堿燒傷后T淋巴細胞介導(dǎo)的特異性免疫反應(yīng),同時也可間接減輕非特異性炎癥反應(yīng),并且表現(xiàn)為濃度依賴性。FK506局部干預(yù)后可阻止角膜堿燒傷后基質(zhì)融解程度,抑制角膜新生血管的生長,減輕角膜組織結(jié)構(gòu)的破壞,改善角膜預(yù)后,從而為角膜堿燒傷急性期的治療提供了新的免疫治療模式。
[Abstract]:Aim: to study the effective concentration and administration of tacrolimus FK506) eye drops in the treatment of corneal alkali burns in rats, to evaluate the efficacy of tacrolimus FK506 eye drops in inhibiting the immune inflammation and improving the corneal microenvironment after alkali burns in rats, and to explore its mechanism. Methods 96 wistar rats with moderate alkali burn of cornea were randomly divided into four groups: group A (n = 4) and group C (n = 4). The group D was treated with FK506 eye drops 4 times per day as the negative control group. The corneal transparency was observed in different groups of rats. The degree of stromal fusion and the formation of neovascularization. At different time points (3 ~ 714 ~ 30 days), rats were killed to remove cornea, and HE staining was used to analyze the changes of corneal tissue structure, and to compare the degree of inflammatory cell infiltration. The temporal and spatial expression of CD4/CD8 positive T cells was detected by immunofluorescence assay. Results: after alkali filter paper burn cornea, corneal haze edema, corneal epithelium loss, matrix fusion ulcer, corneal limbal vasodilation and neovascularization were formed. Corneal loss of transparency. FK506 treatment group in corneal ulcer formation area, Corneal opacity grade in corneal neovascularization was significantly better than that in control group. 1. Quantitative analysis of corneal fusion: the area of corneal fusion in group C was 3.14 鹵0.25 mm ~ 2 鹵2.65 鹵0.28 mm ~ (2) 鹵0.47) mm ~ (2), respectively, compared with that in group D (4.06 鹵0.41) mm ~ (2) (P < 0.05). Significance of P0.05, low concentration of FK506 treatment group and middle, There was also a statistical difference between the high concentration group and the high concentration group, but there was no significant difference between the 0.1% and 0.5% concentration groups, and the area of corneal fusion decreased at 7 days. The results of quantitative analysis of corneal opacification: the score of corneal opacification after burn in the treatment group was lower than that in the control group (P0.05), the rule and the corneal surface of the corneal fusion surface were significantly lower than that in the control group (P 0.05), compared with the control group D (2.47 鹵0.54) mm 2. The corneal opacity score in group A was 1.82 鹵0.51 鹵0.41 鹵0.41 鹵0.41 mm ~ (2) respectively, and that in group C was 0.90 鹵0.39 mm ~ (2) 2, which was significantly different from that in group D (P < 0.05). The change curve of the product is consistent, Results of quantitative analysis of corneal neovascularization: the area of corneal neovascularization in the treatment group was 4.26 鹵0.36 mm ~ (-2) mm in group A on the 3rd day after burn, which was 3.88 鹵0.33 mm ~ (2) mm2 in group B and 3.54 鹵0.46) mm2 in group C, respectively, which was significantly lower than that in group D (5.00 鹵0.12) mm. The difference between P0.01A and BHOC was statistically significant, but there was no significant difference between B and C, and the difference was gradually decreased until the 14th day. The number of inflammatory cells infiltrated in the matrix of FK506 treated group was obviously lower than that of control group by HE staining of burn cornea. The counting results showed that the number of CD4 positive cells in group A was 77.67 鹵4.51) and that in group B was 51.33 鹵3.21) / 0.04mm2m-2 on day 3, while that in group D was 97.00 鹵6.25 / 0.04mm2.The number of CD4 positive cells in group B was lower than that in group B, and the number of positive cells in group B was lower than that in group B (P < 0.05). Medium intensity fluorescence was found in low concentration group, low fluorescence in medium and high concentration group, and diffuse hyperfluorescence in control group. The expression of CD8 T lymphocytes was observed in the whole pathological process of alkali burn of cornea. There was no significant difference between the treatment group and the control group. Conclusion: topical use of tacrolimus FK506 can significantly down-regulate the specific immune response mediated by T lymphocytes after alkali burn of cornea and indirectly attenuate the nonspecific inflammatory response. The results showed that local intervention with concentration dependent. FK506 could prevent the degree of stromal fusion after alkali burn, inhibit the growth of corneal neovascularization, alleviate the destruction of corneal tissue structure, and improve the prognosis of cornea. This provides a new immunotherapy model for the treatment of corneal alkali burn in acute stage.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R779.1

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本文編號：1623875