The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economic...The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.展开更多
目的检测妊娠期糖尿病(GDM)患者血清微小RNA(miR)⁃142⁃3p、miR⁃497⁃5p表达水平,分析其对GDM的诊断价值。方法选取2019年6月-2022年6月在河南中医药大学第五临床医学院妇产科行产前检查的142例GDM孕妇作为GDM组,另外随机选择一般资料与GD...目的检测妊娠期糖尿病(GDM)患者血清微小RNA(miR)⁃142⁃3p、miR⁃497⁃5p表达水平,分析其对GDM的诊断价值。方法选取2019年6月-2022年6月在河南中医药大学第五临床医学院妇产科行产前检查的142例GDM孕妇作为GDM组,另外随机选择一般资料与GDM组匹配的142名正常糖耐量孕妇作为对照组。比较两组孕妇血清miR⁃142⁃3p、miR⁃497⁃5p水平,使用受试者工作特征(ROC)曲线分析血清miR⁃142⁃3p、miR⁃497⁃5p水平对孕妇发生GDM的评估价值。结果与对照组比较,GDM组患者miR⁃142⁃3p水平、有糖尿病史占比、经产妇占比、空腹血糖(FPG)、餐后2 h血糖(2 h PG)、空腹胰岛素(FINS)、糖化血红蛋白(HbA1c)、胰岛素抵抗指数(HOMA⁃IR)升高,血清miR⁃497⁃5p水平显著降低,差异均有统计学意义(t=8.303、6.818、9.551、8.781、9.054、6.464、9.475、11.671、7.883,P均<0.05)。miR⁃142⁃3p、miR⁃497⁃5p联合检测预测孕妇发生GDM的ROC曲线下面积(AUC)为0.838,敏感性为73.24%,特异性为82.39%,优于各自单独诊断(Z=4.379、3.170,P均<0.01)。结论GDM患者血清miR⁃142⁃3p表达显著上调、miR⁃497⁃5p表达显著下调,二者联合对孕妇是否发生GDM有较好的预测价值。展开更多
文摘The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.
文摘目的检测妊娠期糖尿病(GDM)患者血清微小RNA(miR)⁃142⁃3p、miR⁃497⁃5p表达水平,分析其对GDM的诊断价值。方法选取2019年6月-2022年6月在河南中医药大学第五临床医学院妇产科行产前检查的142例GDM孕妇作为GDM组,另外随机选择一般资料与GDM组匹配的142名正常糖耐量孕妇作为对照组。比较两组孕妇血清miR⁃142⁃3p、miR⁃497⁃5p水平,使用受试者工作特征(ROC)曲线分析血清miR⁃142⁃3p、miR⁃497⁃5p水平对孕妇发生GDM的评估价值。结果与对照组比较,GDM组患者miR⁃142⁃3p水平、有糖尿病史占比、经产妇占比、空腹血糖(FPG)、餐后2 h血糖(2 h PG)、空腹胰岛素(FINS)、糖化血红蛋白(HbA1c)、胰岛素抵抗指数(HOMA⁃IR)升高,血清miR⁃497⁃5p水平显著降低,差异均有统计学意义(t=8.303、6.818、9.551、8.781、9.054、6.464、9.475、11.671、7.883,P均<0.05)。miR⁃142⁃3p、miR⁃497⁃5p联合检测预测孕妇发生GDM的ROC曲线下面积(AUC)为0.838,敏感性为73.24%,特异性为82.39%,优于各自单独诊断(Z=4.379、3.170,P均<0.01)。结论GDM患者血清miR⁃142⁃3p表达显著上调、miR⁃497⁃5p表达显著下调,二者联合对孕妇是否发生GDM有较好的预测价值。
