【摘要】：目的探討敲除Toll樣受體2(toll-like receptor 2,TLR2)基因后是否抑制同種異體小鼠角膜排斥反應的發(fā)生。方法以BALB/c為供體,各取30只C57BL/6和同背景TLR2基因敲除鼠為受體行右眼角膜移植術,設為野生組(WT)和基因敲除組(KO);取19只C57BL/6行右眼自體角膜移植術,為自體組(ISO);各取9只C57BL/6和TLR2基因敲除鼠分別設為野生對照組(WT control)和基因敲除對照組(KO control)。術后每周兩次觀察植片并記錄免疫排斥的發(fā)生時間;術后14 d,收集術眼同側頸部淋巴結,行流式細胞術分析CD4+T細胞百分比;收集術眼角膜,行免疫組織化學染色和實時熒光定量PCR檢測角膜干擾素(interferon,IFN)-γ、TLR2和My D88的表達。結果 WT組和KO組小鼠角膜移植術后中位生存時間KO組(35.0±3.8)d長于WT組(21.0±1.5)d(P0.05)。流式細胞術分析顯示,WT組同側頸部淋巴結CD4+T細胞百分比較WT control組明顯增高(P0.05),而ISO和KO組相對于其對應的control組(WT/KO control)差異均無統計學意義(均為P0.05);免疫組織化學結果顯示,ISO和KO組間角膜IFN-γ和My D88分子表達無差異,但兩者均低于WT組。KO組角膜幾乎不表達TLR2分子,ISO組有微量表達,WT組表達明顯增高;PCR檢測顯示,ISO和KO組角膜IFN-γ和TLR2 mRNA相對表達量差異均無統計學意義(均為P0.05),但兩者均低于WT組(均為P0.05);KO組角膜My D88 mRNA相對表達量比ISO組高(P0.05),但兩者均低于WT組(均為P0.05)。結論敲除TLR2基因可以一定程度抑制同種異體小鼠角膜排斥反應的發(fā)生。
[Abstract]:Objective to investigate whether knockout of Toll like receptor 2 (toll-like receptor 2 tLR2) gene inhibits corneal rejection in allogeneic mice. Methods using BALB/c as donor, 30 C57BL/6 and 30 homologous TLR2 knockout mice were used as recipients for right eye keratoplasty, 19 C57BL/6 were divided into wild group (WT) and gene knockout group (KO);) for right eye autogenous keratoplasty. Nine C57BL/6 and TLR2 gene knockout mice were selected as wild control group (WT control) and gene knockout control group (KO control). Respectively for autologous (ISO);. The grafts were observed twice a week and the time of immune rejection was recorded. The ipsilateral cervical lymph nodes were collected on the 14th day after operation, and the percentage of CD4 T cells was analyzed by flow cytometry. The expression of corneal interferon (interferon,IFN)-緯 -tLR2 and My D88 were detected by immunohistochemical staining and real-time fluorescence quantitative PCR. Results the median survival time after corneal transplantation in WT group and KO group was (35.0 鹵3.8) days longer than that in WT group (21.0 鹵1.5) d (P0.05). Flow cytometry analysis showed that the percentage of CD4 T cells in ipsilateral cervical lymph nodes in WT group was significantly higher than that in WT control group (P0.05), but there was no significant difference between ISO and KO group compared with their corresponding control group (WT/KO control) (P0.05). There was no difference in the expression of IFN- 緯 and My D88 between ISO and KO groups. However, the expression of IFN- 緯 and TLR2 mRNA in both groups were significantly higher than those in WT group. KO group showed that there was no significant difference in the relative expression of IFN- 緯 and TLR2 mRNA between ISO group and KO group (P0.05), but there was no significant difference in IFN- 緯 and TLR2 mRNA expression between ISO group and KO group (P0.05). The relative expression of My D88 mRNA in KO group was higher than that in ISO group (P0.05), but both were lower than WT group (P0.05). Conclusion knockout of TLR2 gene can inhibit corneal rejection in allogeneic mice to some extent.
【作者單位】： 南方醫(yī)科大學南方醫(yī)院眼科;
【基金】：國家自然科學基金資助(編號:81170887) 南方醫(yī)科大學南方醫(yī)院橫向課題匹配基金資助(編號:G201202)~~
【分類號】：R779.65

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