目的观察脓毒症并发肝损伤患儿血清高尔基体糖蛋白73(Golgi protein 73, GP73)、miR-122a表达情况,评估其预测脓毒症并发肝损伤患儿预后的价值。方法脓毒症并发肝损伤患儿119例(合并肝损伤组)、单纯脓毒症患儿50例(单纯脓毒症组)和50例...目的观察脓毒症并发肝损伤患儿血清高尔基体糖蛋白73(Golgi protein 73, GP73)、miR-122a表达情况,评估其预测脓毒症并发肝损伤患儿预后的价值。方法脓毒症并发肝损伤患儿119例(合并肝损伤组)、单纯脓毒症患儿50例(单纯脓毒症组)和50例健康儿童(对照组),采用ELISA法检测血清GP73水平,采用实时荧光定量PCR法检测血清miR-122a相对表达量。比较轻度、中度、重度肝损伤脓毒症患儿血清GD73水平和miR-122a相对表达量。合并肝损伤组患儿入院28 d后死亡者30例为死亡组,存活者89例为存活组,比较死亡组和存活组患儿临床资料;采用多因素logistic回归分析脓毒症并发肝损伤患儿不良预后的危险因素;绘制ROC曲线,评估血清GP73水平和miR-122a相对表达量对脓毒症并发肝损伤患儿不良预后的预测价值。结果合并肝损伤组血清GP73水平[(68.53±21.35)μg/L]、miR-122a相对表达量(3.12±0.65)高于单纯脓毒症组[(42.13±11.05)μg/L、2.43±0.46]和对照组[(13.13±6.25)μg/L、1.08±0.21](P<0.05),单纯脓毒症组高于对照组(P<0.05)。轻度、中度、重度肝损伤脓毒症患儿血清GP73水平和miR-122a相对表达量依次增高(P<0.05)。死亡组重度肝损伤患儿比率、APACHEⅡ评分、合并急性肾损伤患儿比率、血清GP73水平和miR-122a相对表达量高于存活组(P<0.05),年龄、性别比例、机械通气比率、ICU住院时间、合并凝血功能障碍比率与存活组比较差异无统计学意义(P>0.05)。重度肝损伤(OR=1.701,95%CI:1.035~3.125,P=0.006)、高APACHEⅡ评分(OR=1.996,95%CI:1.649~3.294,P=0.003)、高GP73水平(OR=2.024,95%CI:1.951~4.099,P<0.001)、高miR-122a相对表达量(OR=2.020,95%CI:1.923~4.094,P<0.001)是脓毒症并发肝损伤患儿不良预后的独立危险因素。当GP73、miR-122a最佳截断值分别为73.26μg/L、3.37时,GP73、miR-122a、GP73+miR-122a预测脓毒症并发肝损伤患儿不良预后的AUC分别为0.767、0.774、0.981,灵敏度分别为70.00%、76.67%、90.00%,特异度分别为80.90%、78.65%、93.26%。结论脓毒症并发肝损伤患儿血清GP73水平和miR-122a相对表达量升高;GP73和miR-122a表达升高是脓毒症并发肝损伤患儿预后不良的危险因素,可作为其预后评估的辅助指标。展开更多
Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the developme...Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the development mechanisms of HCC and devise efficient intervention and treatment protocols.Methods:An upstream miRNA of solute carrier transporter family 1 member 5(SLC1A5)was predicted to bemiR-122-5p by various databases,and a dual-luciferase reporter gene assay was used to verify the SLC1A5-and miR-122-5p-targeting relationship.SLC1A5 and miR-122-5p expression in HCC cells was quantitatively assessed using quantitative reverse transcription polymerase chain reaction(qRT–PCR).Western blotting was used to evaluate SLC1A5 expression in HCC cells.To determine the effects of the ferroptosis inducers erastin and L-g-glutamyl-p-nitroanilide(GPNA)on Huh-7 and HepG2 cell viability,a Cell Counting Kit-8 assay was performed.Additionally,various assay kits were used to assess malondialdehyde(MDA),Fe^(2+),and reactive oxygen species(ROS)levels inHCC cells.Moreover,anHCC tumor model was established in nude mice to investigate the growth of HCC tumors overexpressing miR-122-5p.Results:miR-122-5p downregulated SLC1A5 levels.MiR-122-5p knockdown inhibited erastin-promoted ferroptosis,whereas miR-122-5p overexpression promoted ferroptosis.After knocking down miR-122-5p,the MDA,Fe^(2+),and ROS levels in Huh-7 and HepG2 cells decreased,whereas overexpressing miR-122-5p increased the MDA,Fe^(2+),and ROS levels.Following the addition of GPNA,the cells experienced decreased viability and increased MDA levels,which in turn inhibited ferroptosis.Knockdown of SLC1A5 partially reversed the ferroptosis-inhibiting effect induced by knocking downmiR-122-5p.Additionally,the overexpression of miR-122-5p hindered HCC development in vivo.Conclusion:miR-122-5p downregulates SLC1A5,which promotes lipid peroxidation and iron accumulation and inhibits glutamine transport,ultimately causing ferroptosis in HCC cells.展开更多
文摘目的观察脓毒症并发肝损伤患儿血清高尔基体糖蛋白73(Golgi protein 73, GP73)、miR-122a表达情况,评估其预测脓毒症并发肝损伤患儿预后的价值。方法脓毒症并发肝损伤患儿119例(合并肝损伤组)、单纯脓毒症患儿50例(单纯脓毒症组)和50例健康儿童(对照组),采用ELISA法检测血清GP73水平,采用实时荧光定量PCR法检测血清miR-122a相对表达量。比较轻度、中度、重度肝损伤脓毒症患儿血清GD73水平和miR-122a相对表达量。合并肝损伤组患儿入院28 d后死亡者30例为死亡组,存活者89例为存活组,比较死亡组和存活组患儿临床资料;采用多因素logistic回归分析脓毒症并发肝损伤患儿不良预后的危险因素;绘制ROC曲线,评估血清GP73水平和miR-122a相对表达量对脓毒症并发肝损伤患儿不良预后的预测价值。结果合并肝损伤组血清GP73水平[(68.53±21.35)μg/L]、miR-122a相对表达量(3.12±0.65)高于单纯脓毒症组[(42.13±11.05)μg/L、2.43±0.46]和对照组[(13.13±6.25)μg/L、1.08±0.21](P<0.05),单纯脓毒症组高于对照组(P<0.05)。轻度、中度、重度肝损伤脓毒症患儿血清GP73水平和miR-122a相对表达量依次增高(P<0.05)。死亡组重度肝损伤患儿比率、APACHEⅡ评分、合并急性肾损伤患儿比率、血清GP73水平和miR-122a相对表达量高于存活组(P<0.05),年龄、性别比例、机械通气比率、ICU住院时间、合并凝血功能障碍比率与存活组比较差异无统计学意义(P>0.05)。重度肝损伤(OR=1.701,95%CI:1.035~3.125,P=0.006)、高APACHEⅡ评分(OR=1.996,95%CI:1.649~3.294,P=0.003)、高GP73水平(OR=2.024,95%CI:1.951~4.099,P<0.001)、高miR-122a相对表达量(OR=2.020,95%CI:1.923~4.094,P<0.001)是脓毒症并发肝损伤患儿不良预后的独立危险因素。当GP73、miR-122a最佳截断值分别为73.26μg/L、3.37时,GP73、miR-122a、GP73+miR-122a预测脓毒症并发肝损伤患儿不良预后的AUC分别为0.767、0.774、0.981,灵敏度分别为70.00%、76.67%、90.00%,特异度分别为80.90%、78.65%、93.26%。结论脓毒症并发肝损伤患儿血清GP73水平和miR-122a相对表达量升高;GP73和miR-122a表达升高是脓毒症并发肝损伤患儿预后不良的危险因素,可作为其预后评估的辅助指标。
基金Hubei Provincial Key Research and Development Program(2023BCB126)Hubei University of ChineseMedicine“14th Five-Year Plan”Outstanding Discipline TeamConstruction Project(HBUCM[2022]No.90).
文摘Background:Hepatocellular carcinoma(HCC)typically begins inconspicuously and progresses swiftly,leading to most patients being diagnosed at an advanced stage.Accordingly,a pressing priority is to clarify the development mechanisms of HCC and devise efficient intervention and treatment protocols.Methods:An upstream miRNA of solute carrier transporter family 1 member 5(SLC1A5)was predicted to bemiR-122-5p by various databases,and a dual-luciferase reporter gene assay was used to verify the SLC1A5-and miR-122-5p-targeting relationship.SLC1A5 and miR-122-5p expression in HCC cells was quantitatively assessed using quantitative reverse transcription polymerase chain reaction(qRT–PCR).Western blotting was used to evaluate SLC1A5 expression in HCC cells.To determine the effects of the ferroptosis inducers erastin and L-g-glutamyl-p-nitroanilide(GPNA)on Huh-7 and HepG2 cell viability,a Cell Counting Kit-8 assay was performed.Additionally,various assay kits were used to assess malondialdehyde(MDA),Fe^(2+),and reactive oxygen species(ROS)levels inHCC cells.Moreover,anHCC tumor model was established in nude mice to investigate the growth of HCC tumors overexpressing miR-122-5p.Results:miR-122-5p downregulated SLC1A5 levels.MiR-122-5p knockdown inhibited erastin-promoted ferroptosis,whereas miR-122-5p overexpression promoted ferroptosis.After knocking down miR-122-5p,the MDA,Fe^(2+),and ROS levels in Huh-7 and HepG2 cells decreased,whereas overexpressing miR-122-5p increased the MDA,Fe^(2+),and ROS levels.Following the addition of GPNA,the cells experienced decreased viability and increased MDA levels,which in turn inhibited ferroptosis.Knockdown of SLC1A5 partially reversed the ferroptosis-inhibiting effect induced by knocking downmiR-122-5p.Additionally,the overexpression of miR-122-5p hindered HCC development in vivo.Conclusion:miR-122-5p downregulates SLC1A5,which promotes lipid peroxidation and iron accumulation and inhibits glutamine transport,ultimately causing ferroptosis in HCC cells.
