本文選題：ResolvinE1　切入點(diǎn)：高危角膜移植排斥反應(yīng)　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:觀察在高危角膜移植動物模型中,ResolvinE1(RvE1)對術(shù)后免疫排斥反應(yīng)的抑制及延長角膜植片存活時間的作用。方法:采用隨機(jī)數(shù)字表法將90只BALB/C小鼠分成異體角膜移植+生理鹽水組(Control組)、異體角膜移植+RvE1組(RvE1組)及自體角膜移植組(Syngeneic組),每組各30只。以BALB/c小鼠為受體,C57BL/6小鼠為供體,受體右眼縫線法誘導(dǎo)2周后行穿透性角膜移植手術(shù)。在術(shù)后連續(xù)7天每日對Control組給予不含藥物的生理鹽水10ul結(jié)膜下注射,RvE1組給予ResolvinE1 1ug/10ul,Syngeneic組給予生理鹽水10ul。在裂隙燈下觀察小鼠角膜移植物透明情況并對其排斥反應(yīng)進(jìn)行評分。術(shù)后3周對移植物做HE染色,觀察角膜病理學(xué)改變;采用免疫組化檢測角膜CD4、IFN-γ及IL-17的表達(dá);流式細(xì)胞術(shù)檢測術(shù)眼側(cè)頸部淋巴結(jié)Th1、Th17和Treg細(xì)胞的表達(dá)情況;熒光定量PCR法及CBA檢測Th1、Th2及Th17細(xì)胞相關(guān)因子的表達(dá)水平。結(jié)果:RvE1組與Control組相比,植片存活時間明顯延長,差異有統(tǒng)計學(xué)意義(P0.05)。RvE1組植片水腫程度、炎癥細(xì)胞及CD4+T細(xì)胞(Th1/Th17)浸潤程度較Control組明顯減輕。術(shù)后3周,與Control組相比,RvE1組術(shù)眼測引流淋巴結(jié)中Th1細(xì)胞、Th17細(xì)胞比例明顯減少;植片中IL-1α,IL-1β,TNF-α,IL-2,IL-6,IFN-γ,IL-17A,IL-17F,IL-21和IL-22 mRNA水平明顯降低,IL-2,TNF,IL-6,IFN-γ和IL-17蛋白水平也顯著降低(差異均有統(tǒng)計學(xué)意義)。然而,RvE1組引流淋巴結(jié)中Treg細(xì)胞比例較Control組無明顯改變,IL-4,IL-5和IL-10細(xì)胞因子水平亦無明顯差異。結(jié)論:一、ResolvinE1可抑制高危角膜移植術(shù)后免疫排斥反應(yīng),減少植片水腫及炎癥細(xì)胞的浸潤,延長植片存活時間。二、Resolvin E1可能是通過下調(diào)Th1/Th17細(xì)胞相關(guān)的炎癥反應(yīng)來達(dá)到抑制免疫排斥的作用。三、ResolvinE1可能不是通過上調(diào)Treg細(xì)胞相關(guān)的免疫赦免反應(yīng)來達(dá)到抑制免疫排斥的作用。
[Abstract]:Objective: to observe the inhibitory effect of ResolvinE1 (RvE1) on postoperative immune rejection and prolong the survival time of corneal graft in high risk corneal transplantation animal model. Methods: 90 BALB/C mice were divided into allograft keratoplasty by random number table. Normal saline group, allograft RvE1 group (RvE1 group) and autogenous cornea transplantation group (30 rats in each group). BALB/c mice were used as recipients of C57BL / 6 mice. Penetrating keratoplasty was performed 2 weeks after suture induction in the right eye of the receptor. After 7 days of operation, the Control group was given normal saline 10ul without drugs, and the RvE1 group was given ResolvinE1 1ugr / 10ulu Syngeneic group, and the ResolvinE1 1ugr / 10ulu Syngeneic group was given normal saline 10ul. in the slit-lamp group, the patients in the RVE1 group were treated with normal saline 10ul daily. The grafts were stained with HE at 3 weeks after operation. The expression of IFN- 緯 and IL-17 were detected by immunohistochemistry and Th1T17 and Treg cells in lateral cervical lymph nodes were detected by flow cytometry. Fluorescence quantitative PCR and CBA were used to detect the expression of Th1Th _ 2 and Th17 cell related factors. Results the survival time of grafts in the group of RvE1 was significantly longer than that in the group of Control, and there was significant difference in edema of grafts in the group of P0.05-RvE1. The infiltration degree of inflammatory cells and CD4 T cell Th1 / Th17 was significantly less than that of Control group. 3 weeks after operation, compared with Control group, the percentage of Th1 cell Th17 cells in the drainage lymph nodes of RvE1 group was significantly lower than that in Control group. The levels of IL-17FU IL-21 and IL-22 mRNA of IL-17An IL-17FU IL-21 and IL-22 mRNA in grafts were significantly lower than those in Control group. (all the differences were statistically significant. However, there was no significant change in the percentage of Treg cells in lymph node drainage in RvE1 group compared with Control group, and there was no significant change in IL-4 IL-5 and IL-10 cytokine levels in the RvE1 group, nor did the level of IL-4 IL-5 and IL-10 cytokines in the RvE1 group. The level of IL-4 IL-5 and IL-10 cytokines in the RvE1 group was not significantly changed compared with that in the Control group. Conclusion: ResolvinE1 can inhibit the immune rejection after high risk corneal transplantation. Reduce graft edema and inflammatory cell infiltration, It is suggested that the inhibition of immune rejection may be achieved by down-regulating the inflammatory response of Th1/Th17 cells, while ResolvinE1 may not inhibit immune rejection by upregulating the immune amnesty response associated with Treg cells.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R779.65

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