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IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究

孟强, 张文杰, 吴伟康, 等. IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究[J]. 器官移植, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
引用本文: 孟强, 张文杰, 吴伟康, 等. IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究[J]. 器官移植, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
Meng Qiang, Zhang Wenjie, Wu Weikang, et al. Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models[J]. ORGAN TRANSPLANTATION, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010
Citation: Meng Qiang, Zhang Wenjie, Wu Weikang, et al. Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models[J]. ORGAN TRANSPLANTATION, 2023, 14(1): 75-82. doi: 10.3969/j.issn.1674-7445.2023.01.010

IL-18/IL-18BP介导NK-92MI细胞杀伤GTKO猪内皮细胞的初步研究

doi: 10.3969/j.issn.1674-7445.2023.01.010
基金项目: 

国家自然科学基金 82170667

详细信息
    作者简介:
    通讯作者:

    陶开山,博士,主任医师,教授,研究方向为异种肝移植,肝癌,Email: taokaishan0686@163.com

  • 中图分类号: R617

Preliminary study of the role of IL-18/IL-18BP in mediating cytotoxic ability of NK-92MI cells against endothelial cells from GTKO porcine models

More Information
  • 摘要:   目的  探讨白细胞介素(IL)-18/IL-18结合蛋白(BP)介导自然杀伤(NK)-92MI细胞杀伤α-1, 3-半乳糖基转移酶(GGTA1)基因敲除(GTKO)猪内皮细胞的效应及可能机制。  方法  将NK-92MI细胞分为NK组、NK+IL-18组、NK+GTKO组、IL-18+NK+GTKO组和IL-18+IL-18BP+NK+GTKO组,采用实时定量逆转录聚合酶链反应(qRT-PCR)检测NK-92MI细胞中炎症相关基因的信使核糖核酸(mRNA)表达,乳酸脱氢酶(LDH)法检测NK-92MI细胞对GTKO猪内皮细胞的杀伤效应,脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记(TUNEL)法检测GTKO猪内皮细胞的凋亡情况,蛋白质印迹法检测具有杀伤效应的蛋白和凋亡相关蛋白的表达。  结果  与NK组、NK+IL-18组以及NK+GTKO组比较,IL-18+NK+GTKO组NK-92MI细胞中干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、IL-8、IL-3、IL-6和粒细胞-巨噬细胞集落刺激因子(GM-CSF)的mRNA表达水平均升高,差异均有统计学意义(均为P < 0.05)。与IL-18+NK+GTKO组比较,IL-18+IL-18BP+NK+GTKO组NK-92MI细胞中IFN-γ、TNF-α、IL-8、IL-3、IL-6和GM-CSF的mRNA表达水平均降低,差异均有统计学意义(均为P < 0.05)。与NK+GTKO组相比,IL-18+NK+GTKO组NK-92MI细胞中穿孔素、颗粒酶B和IFN-γ蛋白表达水平均升高,NK-92MI细胞对GTKO猪内皮细胞的杀伤率增加,GTKO猪内皮细胞凋亡率增加,GTKO猪内皮细胞中B淋巴细胞瘤-2(Bcl-2)相关X蛋白(Bax)/Bcl-2和裂解半胱氨酸天冬氨酸蛋白酶(cleaved Caspase)-3/半胱氨酸天冬氨酸蛋白酶(Caspase)-3的比例均升高,差异均有统计学意义(均为P < 0.05)。与IL-18+NK+GTKO组比较,IL-18+IL-18BP+NK+GTKO组穿孔素、颗粒酶B、IFN-γ蛋白表达水平均降低,NK-92MI细胞对GTKO猪内皮细胞的杀伤率降低,GTKO猪内皮细胞凋亡率下降,GTKO猪内皮细胞中Bax/Bcl-2和cleaved Caspase-3/Caspase-3的比例均下降,差异均有统计学意义(均为P < 0.05)。  结论  IL-18BP可阻断IL-18诱导的NK-92MI细胞中炎症相关基因表达和NK-92MI细胞杀伤GTKO猪内皮细胞的效应。

     

  • 图  1  各组NK-92MI细胞中炎症相关基因的表达

    注:与NK组比较,aP < 0.05;与NK+IL-18组比较,bP < 0.05;与NK+GTKO组比较,cP < 0.05;与IL-18+NK+GTKO组比较,dP < 0.05。

    Figure  1.  Expression of inflammation-related genes of NK-92MI cells in each group

    图  2  各组NK-92MI细胞中杀伤效应蛋白的表达

    注:与NK+GTKO组比较,aP < 0.05;与IL-18+NK+GTKO组比较,bP < 0.05。

    Figure  2.  Expression of killer effector proteins of NK-92MI cells in each group

    图  3  各组NK-92MI细胞对GTKO猪内皮细胞的杀伤效应

    注:与NK+GTKO组比较,aP < 0.05;与IL-18+NK+GTKO组比较,bP < 0.05。

    Figure  3.  Cytotoxic effect of NK-92MI cells on GTKO porcine endothelial cells in each group

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  • 收稿日期:  2022-11-14
  • 网络出版日期:  2023-01-17
  • 刊出日期:  2023-01-15

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