ApoE基因对老年小鼠脊髓组织炎症因子表达的影响

Effect of ApoE Gene on the Expression of Inflammatory Factors in Spinal Cord Tissue of Aged Mice

目的探讨载脂蛋白E(apolipoprotein E,ApoE)基因对老年小鼠脊髓组织炎症因子表达的影响。方法培养C57/BL6野生型小鼠和ApoE基因敲除小鼠,饲养至16~18个月,建立自然衰老模型。取材小鼠脊髓组织,应用免疫组织化学染色技术观察白介素6(interleukin 6,IL-6)和白介素10(interleukin 10,IL-10)的表达并分析表达量。通过酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测脊髓组织中IL-10和白介素1β(interleukin 1β,IL-1β)的含量。应用实时定量聚合酶链反应(real-time quantitative PCR,RT-qPCR)技术检测肿瘤坏死因子(tumor necrosis factor,TNF)和精氨酸酶1(arginase-1,Arg-1)mRNA的表达。结果免疫组织化学染色、ELISA和RT-qPCR实验结果显示与野生型小鼠比较,老年ApoE基因敲除小鼠脊髓组织中促炎因子IL-6、IL-1β和TNF表达增加,抗炎因子IL-10和Arg-1表达减少。结论ApoE基因影响衰老小鼠脊髓组织炎症因子的表达,该基因可作为老年脊髓疾病治疗的靶点。

Objective To investigate the effect of apolipoprotein E(ApoE)gene on the expression of inflammatory factors in the spinal cord tissue of aged mice.Methods C57/BL6 wild-type mice and ApoE gene knockout mice were cultured for 16-18 months to establish natural aging model.The spinal cord tissues of mice were taken,and the expressions of interleukin 6(IL-6)and interleukin 10(IL-10)were observed and analyzed by immunohistochemical staining.The contents of IL-10 and interleukin 1β(IL-1β)in spinal cord tissue were detected by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative PCR(RT-qPCR)has been used to detect the mRNA expression of tumor necrosis factor(TNF)and arginase-1(Arg-1).Results Immunohistochemical staining,ELISA and RT-qPCR showed that the expressions of pro-inflammatory factors IL-6,IL-1βand TNF were increased,and the expressions of anti-inflammatory factors IL-10 and Arg-1 were decreased in the spinal cord of aged ApoE knockout mice compared with wild-type mice.Conclusion ApoE gene affects the expression of inflammatory cytokines in the spinal cord tissue of senile mice and can be used as a therapeutic target for senile spinal cord diseases.