MCP-1基因对脓毒症急性肾损伤发生的作用研究  

Effect of MCP-1 gene on occurrence of acute kidney injury(AKI)in sepsis

作　　者：郭峻氚[1] 王静静[1] 郭仁楠 肖东[1] 刘艳[1] GUO Junchuan;WANG Jingjing;GUO Rennan;XIAO Dong;LIU Yan(Department of Critical Medicine,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,China)

机构地区：[1]新疆维吾尔自治区人民医院重症医学科,乌鲁木齐830000

出　　处：《新疆医科大学学报》2024年第4期512-517,共6页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区自然科学基金项目(2021D01C156)。

摘　　要：目的研究单核细胞趋化蛋白-1(MCP-1)基因对脓毒症急性肾损伤(AKI)发生的作用。方法选取2022年9月-2023年9月新疆维吾尔自治区人民医院重症医学科收治的50例脓毒症AKI患者作为AKI组,纳入同期50例健康受试者作为对照组。分别采集全部受试者清晨空腹静脉血5 mL,采用ELISA法测定AKI患者及健康人群血清中MCP-1的表达情况;通过原代培养肾小管上皮细胞,利用CK14和CK18抗体进行细胞免疫荧光鉴定,过表达和干扰MCP-1基因,利用CCK8细胞增殖检测试剂盒和RT-qPCR检测肾小管上皮细胞增殖情况和肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素6(IL-6)、白细胞介素10(IL-10)、γ-干扰素(IFN-γ)炎性因子表达的变化。结果与对照组相比,AKI组患者外周血中和尿液中的MCP-1表达量显著升高(P均<0.05);通过细胞免疫荧光鉴定,选择上皮细胞标志物CK14和CK18,原代培养24 h,90%以上的细胞表达细胞标志物CK18,约84%的细胞表达CK14;与NC组相比,siRNA组在24、48 h细胞数量增加(P均<0.05);与MCP-1组相比,siRNA组在24、48、72 h的细胞数量增加(P均<0.05);NC组的IL-1β、IL-6、INF-γ因子的表达水平随时间推移逐渐升高,MCP-1组的IL-1β、IL-6、IL-10、INF-γ和TNF-α因子的表达水平随时间推移逐渐升高,siRNA组的IL-1β、IL-6、INF-γ因子表达水平随时间推移无明显升高趋势。结论MCP-1基因在脓毒症急性肾损伤的发病机制中可能发挥重要作用,该基因可能通过调节IL-1β等炎性细胞因子的表达来抑制肾小管上皮细胞的生长和增殖,从而参与脓毒症患者的AKI过程。Objective To investigate the effect of monocyte chemoattractant protein-1(MCP-1)gene on sepsis induced acute kidney injury(AKI).Methods 50 patients with sepsis AKI admitted to the hospital from September 2022 to September 2023 were selected as AKI group,and 50 healthy subjects in the same period were included as control group.5 mL fasting venous blood was collected from all subjects in the morning,and the expression of MCP-1 in the serum of AKI patients and healthy people was detected by ELISA.The primary cultured renal tubular epithelial cells were identified by immunofluorescence using CK14 and CK18 antibodies.MCP-1 gene was over-expressed and interfered.The cell proliferation assay kit and RT-qPCR were used to detect the proliferation of renal tubular epithelial cells and the changes in the expression of inflammatory factors,including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10)andγ-interferon(IFN-γ).Results Compared with control group,the expression of MCP-1 in peripheral blood and urine of AKI group was significantly increased(P<0.05).According to the identification of cellular immunofluorescence,the epithelial cell markers CK14 and CK18 were selected.When cultured for 24 hours,more than 90%of the cells expressed the cell marker CK18,and about 84%of the cells expressed CK14.Compared with NC group,the number of cells in siRNA group were increased at 24 and 48 hours(P<0.05).Compared with MCP-1 group,the number of cells in siRNA group were increased at 24,48 and 72 hours(P<0.05).The expression levels of IL-1β,IL-6 and IFN-γin NC group were increased gradually with the passage of time.The expression levels of IL-1β,IL-6,IL-10,IFN-γand TNF-αin MCP-1 group were increased gradually with the passage of time.The expression levels of IL-1β,IL-6 and IFN-γwere no significant upward trend in siRNA group.Conclusion MCP-1 gene may play an important role in the pathogenesis of sepsis induced AKI,which may regulate IL-1βand other inflammatory cytokines to inhibi

关 键 词：MCP-1基因 脓毒症 肾小管上皮细胞 炎症反应 

分 类 号：R631[医药卫生—外科学]