CRNDE调节miR-126-5p减弱患儿脓毒症脑损伤的机制研究  

The action mechanism of CRNDE alleviating the brain damage in children with sepsis by regulating microRNA-126-5p

作　　者：张晶[1] 代思华 王晓岩 谷丽彩 黄蕊[1] 任常军[1] ZHANG Jing;DAI Sihua;WANG Xiaoyan(Department of Pediatrics,The First Hospital of Hebei Medical University,Hebei,Shijiazhuang 050031,China)

机构地区：[1]河北医科大学第一医院儿科,石家庄市050031

出　　处：《河北医药》2021年第14期2101-2105,共5页Hebei Medical Journal

基　　金：河北省科技厅河北省重点研发计划项目健康医疗与生物医药专项(编号:182777109D);河北医科大学第一医院“星火”青年科研项目(编号:XH201712)。

摘　　要：目的探讨长链非编码RNA(long noncoding RNA)结直肠肿瘤差异表达(colorectal neoplasia differentially expressed,CRNDE)和微小RNA-126-5p(miR-126-5p)在脓毒症相关性脑损伤中的作用机制。方法选择符合脓毒症诊断标准同时无中枢神经系统疾病30例患者作为研究对象,在体外构建LPS诱导的星形胶质细胞模型。采用RT-PCR检测脓毒症患儿血清和LPS诱导的星形胶质细胞中CRNDE和miR-126-5p的水平;采用生物信息法和荧光素酶法推测和证实二者间的相互关系;MTT法检测细胞的增殖;western blot检测凋亡相关蛋白(Bax、Bcl2)、NF-κB、Keap1、Nrf2的水平;采用ELISA检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、超氧化物歧化酶(SOD)、丙二醛(MDA)的水平。结果LPS显著降低了星形胶质细胞细胞中CRNDE的表达水平(P<0.05)。在LPS诱导的星形胶质细胞中过表达CRNDE后,星形胶质细胞的增殖水平上调(P<0.05)。上调LPS诱导的星形胶质细胞中CRNDE的水平后,细胞中Bax的水平显著下调,而Bcl2的水平上调。ELISA和western blot结果显示,上调CRNDE后,LPS介导的星形胶质细胞中TNF-α、IL-1β、NF-κB、MDA、Keap1的水平下调,而SOD和Nrf2的水平上调(P<0.05)。双荧光素酶报告试验表明,miR-126-5p模拟物可降低含CRNDE-wt的荧光素酶报告体的荧光素酶活性,而对CRNDE-mut载体的荧光素酶活性没有显著影响。CRNDE与miR-126-5p相互作用且能负向调节其表达。结论CRNDE能够靶向miR-126-5p减弱脓毒症患儿脑损伤。Objective To investigate the action mechanism of colorectal neoplasia differentially expressed(CRNDE)and microRNA-126-5p(miR-126-5p)in the pathogenesis of sepsis related brain injury in long chain non coding RNA(long noncoding RNA)colorectal cancer.Methods Thirty patients with sepsis and without central nervous system diseases were enrolled in the study,and the LPS-induced astrocyte models were established in vitro.The levels of CRNDE and miR-126-5p in serum and LPS-induced astrocytes were detected by RT-PCR.Bioinformatics and luciferase methods were used to analyze the correlation between them.MTT method was used to detect cell proliferation,Western Blot was used to detect the levels of apoptotic related proteins(Bax,Bcl2),NF-kappa B,Keap1,Nrf2,moreover,ELISA was used to detect the levels of TNF-α,IL-1beta,SOD,MDA.Results The expression levels of CRNDE were down-regulated in astrocyte,however,the overexpression of CRNDE induced by LPS up-regulated the expression levels of mir-126-5p(P<0.05).After up-regulating the expression levels of CRNDE induced by LPS in astrocytes,the Baxlevels were decreased,however,the Bcl2levels were increased.The ELISA and Western Blot results showed that after up-regulating the expression levels of CRNDE,the levels of TNF-α,IL-1β,NF-κB,MDA,Keap1were decreased,but the levels of SOD and Nrf2 were increased significantly(P<0.05).In addition the CRNDE interacts could negatively regulated its expression.Conclusion CRNDE can target RNA-126-5p to alleviate the brain damage in children with sepsis.

关 键 词：CRNDE miR-126-5p 脓毒症介导的脑损伤 增殖 凋亡 

分 类 号：R631[医药卫生—外科学]