AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)...AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.展开更多
Background:Intracerebral hemorrhage(ICH)remains a devastating neurological disorder with limited therapeutic options.Neural stem cell(NSC)-based therapies have emerged as a potential regenerative approach,yet the mole...Background:Intracerebral hemorrhage(ICH)remains a devastating neurological disorder with limited therapeutic options.Neural stem cell(NSC)-based therapies have emerged as a potential regenerative approach,yet the molecular mechanisms regulating NSC behavior require further elucidation.The role of miR-21 in NSC differentiation and proliferation during ICH recovery remains unexplored.Methods:In vitro NSC cultures were analyzed for miR-21 expression dynamics during differentiation via qPCR.Lentiviral overexpression and knockdown of miR-21 were employed to assess its functional impact.The SOX2/LIN28-let-7 pathway was investigated using Western blot,luciferase reporter assays,and immunofluorescence.In vivo,miR-21-overexpressing NSCs were transplanted into a murine ICH model,with neurogenesis evaluated by immunostaining and neurological recovery assessed through behavioral tests(mNSS,rotarod).Results:miR-21 expression significantly increased during NSC differentiation,correlating with reduced SOX2 levels.Mechanistically,miR-21 directly targeted SOX2,disrupting the SOX2/LIN28-let-7 axis to promote NSC proliferation and lineage commitment.In ICH mice,transplantation of miR-21-overexpressing NSCs enhanced neurogenesis and improved motor coordination and neurological deficits at 28 days post-transplantation.Conclusions:Our findings identify miR-21 as a critical regulator of NSC plasticity through SOX2/LIN28-let-7 signaling,highlighting its therapeutic potential for enhancing neuroregeneration and functional recovery in ICH.Targeting miR-21 may represent a novel strategy to optimize NSC-based therapies for hemorrhagic stroke.展开更多
目的探究妊娠期糖尿病患者血清微小RNA(miRNA)-21、miRNA-31水平与凝血功能、炎症反应及妊娠结局的关系。方法回顾性选取2018年2月至2024年4月在邢台市中心医院进行产检并生产的378例妊娠期糖尿病患者为研究对象,并根据其妊娠结局进行分...目的探究妊娠期糖尿病患者血清微小RNA(miRNA)-21、miRNA-31水平与凝血功能、炎症反应及妊娠结局的关系。方法回顾性选取2018年2月至2024年4月在邢台市中心医院进行产检并生产的378例妊娠期糖尿病患者为研究对象,并根据其妊娠结局进行分组:妊娠结局不良组(n=58)和妊娠结局正常组(n=320)。比较组间的血糖指标[口服葡萄糖耐量试验1小时血糖(1 h OGTT)、2 h OGTT、糖化血红蛋白(GHb)、胰岛素抵抗指数(HOMA-IR)]、凝血指标[凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原(FIB)、D-二聚体(D-D)]、炎症反应[脂肪酸结合蛋白4(FABP4)、白细胞介素6(IL-6)、骨保护素、脂联素]及其血清miRNA-21、miRNA-31水平,并探究miRNA-21、miRNA-31水平与其凝血指标、炎症指标的关系。结果两组患者的空腹OGTT、1 h OGTT、2 h OGTT、GHb、HOMA-IR比较,差异均无统计学意义(P>0.05)。妊娠结局不良组患者的PT、APTT分别为(10.31±1.12)、(23.66±2.46)s,均低于妊娠结局正常组[(11.66±2.45)、(27.99±4.15)s],FIB、D-D分别为(4.91±0.46)g/L、(0.98±0.12)μg/L,均高于妊娠结局正常组[(3.88±0.54)g/L、(0.81±0.08)μg/L],差异均有统计学意义(P<0.05)。妊娠结局不良组患者的FABP4、IL-6、骨保护素水平分别为(6.45±1.26)、(361.03±18.46)、0.04(0.02,0.06)pg/L,均高于妊娠结局正常组[(4.33±1.16)、(344.15±22.15)、0.02(0.01,0.04)pg/L],脂联素水平为0.02(0.01,0.04)pg/L,低于妊娠结局正常组[0.03(0.01,0.04)pg/L],差异均有统计学意义(P<0.05)。妊娠结局不良组患者的血清miRNA-21为0.78±0.13,低于妊娠结局正常组(1.23±0.33),miRNA-31水平为2.44±0.66,高于妊娠结局正常组(1.56±0.51),差异均有统计学意义(P<0.05)。经相关性分析,miRNA-21与PT、APTT、脂联素均呈正相关(P<0.05),与FIB、D-D、FABP4、IL-6和骨保护素均呈负相关(P<0.05)。miRNA-31与PT、APTT和脂联素均呈负相关(P<0.05),与FIB、D-D、FABP4、IL-6和骨保护素均呈正相关(P<0.05)。结论在妊娠期糖尿病患者中,妊娠结局不良者血清miRNA-21水平降低、miRNA-31水平升高,妊娠结局与其miRNA-21、miRNA-31存在显著相关性,且二者与其凝血功能、炎症反应指标呈显著相关性。展开更多
文摘AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.
基金Shanghai Minhang District Health Commission Project,Grant/Award Number:2022MHZ062Shanghai Sixth People's Hospital,Grant/Award Number:ynhg202311 and ynms202411。
文摘Background:Intracerebral hemorrhage(ICH)remains a devastating neurological disorder with limited therapeutic options.Neural stem cell(NSC)-based therapies have emerged as a potential regenerative approach,yet the molecular mechanisms regulating NSC behavior require further elucidation.The role of miR-21 in NSC differentiation and proliferation during ICH recovery remains unexplored.Methods:In vitro NSC cultures were analyzed for miR-21 expression dynamics during differentiation via qPCR.Lentiviral overexpression and knockdown of miR-21 were employed to assess its functional impact.The SOX2/LIN28-let-7 pathway was investigated using Western blot,luciferase reporter assays,and immunofluorescence.In vivo,miR-21-overexpressing NSCs were transplanted into a murine ICH model,with neurogenesis evaluated by immunostaining and neurological recovery assessed through behavioral tests(mNSS,rotarod).Results:miR-21 expression significantly increased during NSC differentiation,correlating with reduced SOX2 levels.Mechanistically,miR-21 directly targeted SOX2,disrupting the SOX2/LIN28-let-7 axis to promote NSC proliferation and lineage commitment.In ICH mice,transplantation of miR-21-overexpressing NSCs enhanced neurogenesis and improved motor coordination and neurological deficits at 28 days post-transplantation.Conclusions:Our findings identify miR-21 as a critical regulator of NSC plasticity through SOX2/LIN28-let-7 signaling,highlighting its therapeutic potential for enhancing neuroregeneration and functional recovery in ICH.Targeting miR-21 may represent a novel strategy to optimize NSC-based therapies for hemorrhagic stroke.
文摘目的探究妊娠期糖尿病患者血清微小RNA(miRNA)-21、miRNA-31水平与凝血功能、炎症反应及妊娠结局的关系。方法回顾性选取2018年2月至2024年4月在邢台市中心医院进行产检并生产的378例妊娠期糖尿病患者为研究对象,并根据其妊娠结局进行分组:妊娠结局不良组(n=58)和妊娠结局正常组(n=320)。比较组间的血糖指标[口服葡萄糖耐量试验1小时血糖(1 h OGTT)、2 h OGTT、糖化血红蛋白(GHb)、胰岛素抵抗指数(HOMA-IR)]、凝血指标[凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原(FIB)、D-二聚体(D-D)]、炎症反应[脂肪酸结合蛋白4(FABP4)、白细胞介素6(IL-6)、骨保护素、脂联素]及其血清miRNA-21、miRNA-31水平,并探究miRNA-21、miRNA-31水平与其凝血指标、炎症指标的关系。结果两组患者的空腹OGTT、1 h OGTT、2 h OGTT、GHb、HOMA-IR比较,差异均无统计学意义(P>0.05)。妊娠结局不良组患者的PT、APTT分别为(10.31±1.12)、(23.66±2.46)s,均低于妊娠结局正常组[(11.66±2.45)、(27.99±4.15)s],FIB、D-D分别为(4.91±0.46)g/L、(0.98±0.12)μg/L,均高于妊娠结局正常组[(3.88±0.54)g/L、(0.81±0.08)μg/L],差异均有统计学意义(P<0.05)。妊娠结局不良组患者的FABP4、IL-6、骨保护素水平分别为(6.45±1.26)、(361.03±18.46)、0.04(0.02,0.06)pg/L,均高于妊娠结局正常组[(4.33±1.16)、(344.15±22.15)、0.02(0.01,0.04)pg/L],脂联素水平为0.02(0.01,0.04)pg/L,低于妊娠结局正常组[0.03(0.01,0.04)pg/L],差异均有统计学意义(P<0.05)。妊娠结局不良组患者的血清miRNA-21为0.78±0.13,低于妊娠结局正常组(1.23±0.33),miRNA-31水平为2.44±0.66,高于妊娠结局正常组(1.56±0.51),差异均有统计学意义(P<0.05)。经相关性分析,miRNA-21与PT、APTT、脂联素均呈正相关(P<0.05),与FIB、D-D、FABP4、IL-6和骨保护素均呈负相关(P<0.05)。miRNA-31与PT、APTT和脂联素均呈负相关(P<0.05),与FIB、D-D、FABP4、IL-6和骨保护素均呈正相关(P<0.05)。结论在妊娠期糖尿病患者中,妊娠结局不良者血清miRNA-21水平降低、miRNA-31水平升高,妊娠结局与其miRNA-21、miRNA-31存在显著相关性,且二者与其凝血功能、炎症反应指标呈显著相关性。
