敲除TLR2基因对同种异体小鼠角膜移植排斥反应的影响
发布时间：2018-08-31 16:06
【摘要】：目的探讨敲除Toll样受体2(toll-like receptor 2,TLR2)基因后是否抑制同种异体小鼠角膜排斥反应的发生。方法以BALB/c为供体,各取30只C57BL/6和同背景TLR2基因敲除鼠为受体行右眼角膜移植术,设为野生组(WT)和基因敲除组(KO);取19只C57BL/6行右眼自体角膜移植术,为自体组(ISO);各取9只C57BL/6和TLR2基因敲除鼠分别设为野生对照组(WT control)和基因敲除对照组(KO control)。术后每周两次观察植片并记录免疫排斥的发生时间;术后14 d,收集术眼同侧颈部淋巴结,行流式细胞术分析CD4+T细胞百分比;收集术眼角膜,行免疫组织化学染色和实时荧光定量PCR检测角膜干扰素(interferon,IFN)-γ、TLR2和My D88的表达。结果 WT组和KO组小鼠角膜移植术后中位生存时间KO组(35.0±3.8)d长于WT组(21.0±1.5)d(P0.05)。流式细胞术分析显示,WT组同侧颈部淋巴结CD4+T细胞百分比较WT control组明显增高(P0.05),而ISO和KO组相对于其对应的control组(WT/KO control)差异均无统计学意义(均为P0.05);免疫组织化学结果显示,ISO和KO组间角膜IFN-γ和My D88分子表达无差异,但两者均低于WT组。KO组角膜几乎不表达TLR2分子,ISO组有微量表达,WT组表达明显增高;PCR检测显示,ISO和KO组角膜IFN-γ和TLR2 mRNA相对表达量差异均无统计学意义(均为P0.05),但两者均低于WT组(均为P0.05);KO组角膜My D88 mRNA相对表达量比ISO组高(P0.05),但两者均低于WT组(均为P0.05)。结论敲除TLR2基因可以一定程度抑制同种异体小鼠角膜排斥反应的发生。
[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)-纬 -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 卤3.8) days longer than that in WT group (21.0 卤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- 纬 and My D88 between ISO and KO groups. However, the expression of IFN- 纬 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- 纬 and TLR2 mRNA between ISO group and KO group (P0.05), but there was no significant difference in IFN- 纬 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.
【作者单位】： 南方医科大学南方医院眼科;
【基金】：国家自然科学基金资助(编号:81170887) 南方医科大学南方医院横向课题匹配基金资助(编号:G201202)~~
【分类号】：R779.65
本文编号：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)-纬 -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 卤3.8) days longer than that in WT group (21.0 卤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- 纬 and My D88 between ISO and KO groups. However, the expression of IFN- 纬 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- 纬 and TLR2 mRNA between ISO group and KO group (P0.05), but there was no significant difference in IFN- 纬 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.
【作者单位】： 南方医科大学南方医院眼科;
【基金】：国家自然科学基金资助(编号:81170887) 南方医科大学南方医院横向课题匹配基金资助(编号:G201202)~~
【分类号】：R779.65
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