敲除TLR2基因?qū)νN異體小鼠角膜移植排斥反應(yīng)的影響
發(fā)布時(shí)間：2018-08-31 16:06
【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(yīng)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(shù),設(shè)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(shù),為自體組(ISO);各取9只C57BL/6和TLR2基因敲除鼠分別設(shè)為野生對(duì)照組(WT control)和基因敲除對(duì)照組(KO control)。術(shù)后每周兩次觀察植片并記錄免疫排斥的發(fā)生時(shí)間;術(shù)后14 d,收集術(shù)眼同側(cè)頸部淋巴結(jié),行流式細(xì)胞術(shù)分析CD4+T細(xì)胞百分比;收集術(shù)眼角膜,行免疫組織化學(xué)染色和實(shí)時(shí)熒光定量PCR檢測(cè)角膜干擾素(interferon,IFN)-γ、TLR2和My D88的表達(dá)。結(jié)果 WT組和KO組小鼠角膜移植術(shù)后中位生存時(shí)間KO組(35.0±3.8)d長(zhǎng)于WT組(21.0±1.5)d(P0.05)。流式細(xì)胞術(shù)分析顯示,WT組同側(cè)頸部淋巴結(jié)CD4+T細(xì)胞百分比較WT control組明顯增高(P0.05),而ISO和KO組相對(duì)于其對(duì)應(yīng)的control組(WT/KO control)差異均無統(tǒng)計(jì)學(xué)意義(均為P0.05);免疫組織化學(xué)結(jié)果顯示,ISO和KO組間角膜IFN-γ和My D88分子表達(dá)無差異,但兩者均低于WT組。KO組角膜幾乎不表達(dá)TLR2分子,ISO組有微量表達(dá),WT組表達(dá)明顯增高;PCR檢測(cè)顯示,ISO和KO組角膜IFN-γ和TLR2 mRNA相對(duì)表達(dá)量差異均無統(tǒng)計(jì)學(xué)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(duì)表達(dá)量比ISO組高(P0.05),但兩者均低于WT組(均為P0.05)。結(jié)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(yīng)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(xué)南方醫(yī)院眼科;
【基金】：國家自然科學(xué)基金資助(編號(hào):81170887) 南方醫(yī)科大學(xué)南方醫(yī)院橫向課題匹配基金資助(編號(hào):G201202)~~
【分類號(hào)】：R779.65
本文編號(hào)：2215491
[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(xué)南方醫(yī)院眼科;
【基金】：國家自然科學(xué)基金資助(編號(hào):81170887) 南方醫(yī)科大學(xué)南方醫(yī)院橫向課題匹配基金資助(編號(hào):G201202)~~
【分類號(hào)】：R779.65
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