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
目的探讨茵陈蒿汤对代谢相关脂肪性肝病(MAFLD)小鼠LncMEG3/miR-21-5p/PPARα介导脂肪酸β氧化的影响。方法为探讨MAFLD差异表达的miRNAs,应用PubMed、Web of Science和中国知网3个电子数据库对以中英文发表的研究进行检索,采用随机效...目的探讨茵陈蒿汤对代谢相关脂肪性肝病(MAFLD)小鼠LncMEG3/miR-21-5p/PPARα介导脂肪酸β氧化的影响。方法为探讨MAFLD差异表达的miRNAs,应用PubMed、Web of Science和中国知网3个电子数据库对以中英文发表的研究进行检索,采用随机效应模型进行Meta分析。24只C57BL/6J小鼠,SPF级,采用随机数字表法将小鼠随机分为对照组、模型组和茵陈蒿汤组,共3组,每组8只。采用喂饲高脂饲料的方法对模型组和茵陈蒿汤组小鼠建立MAFLD模型。12周后,茵陈蒿汤组给予茵陈蒿汤灌胃,每日1次,连续4周。采用HE染色法和油红O染色法观察各组小鼠肝组织病理形态表现;全自动生化分析仪检测血清甘油三酯(TG)、总胆固醇(TC)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、高密度脂蛋白胆固(HDL-C)和低密度脂蛋白胆固醇(LDL-C);试剂盒检测小鼠肝脏甘油三酯(TG)和游离脂肪酸(FFA)水平;RT-qPCR法检测小鼠LncMEG3、miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p、过氧化物酶体增殖物激活受体α(PPARα)、肉碱棕榈酰转移酶1A(CPT1A)、肉碱棕榈酰基转移酶2(CPT2)、中链酰基辅酶A脱氢酶(MCAD)、长链脂酰辅酶A脱氢酶(LCAD)、酰基辅酶A氧化酶1(ACOX1)和三羟酰辅酶A脱氢酶(EHHADH)mRNA水平蛋白表达水平。结果Meta分析结果显示,与健康对照组相比,MAFLD组miR-34a-5p、miR-122-5p、miR-192-5p、miR-21-5p、miR-451-5p水平显著升高。肝脏HE染色结果发现,模型组小鼠出现肝细胞肿胀,体积增大,有明显脂肪变性;与模型组相比,茵陈蒿汤组小鼠肝细胞体积肿胀明显降低,脂肪变性显著减少。肝脏油红O染色结果发现,与对照组相比,模型组肝脏可见大量橘红色脂滴,脂质沉积显著升高;茵陈蒿汤组可见少量脂肪变性,橘红色脂滴分布减少,肝脏脂质沉积情况明显改善。相较于对照组,模型组小鼠血清HDL-C、LDL-C、TG、TC、AST、ALT表达显著升高(P<0.05),肝脏TG和FFA水平均显著升高(P<0.05);相较于模型组,茵陈蒿汤组小鼠肝脏FFA和TG水平显著降低(P<0.05),血清AST、LDL-C、TG、ALT水平显著降低(P<0.05)。与对照组相比,模型组miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p表达水平增高(P<0.05);与模型组相比,茵陈蒿汤组miR-21-5p、miR-34a-5p、miR-122-5p、miR-192-5p表达水平降低(P<0.05),miR-21-5p变化倍数(模型组/茵陈蒿汤组)最大。与对照组相比,模型组Lnc MEG3表达水平降低(P<0.05);与模型组相比,茵陈蒿汤组Lnc MEG3表达水平升高(P<0.05)。与对照组相比,模型组PPARα、CPT1A、CPT2、MCAD、LCAD、ACOX1、EHHADH mRNA和蛋白表达水平显著降低(P<0.05);与模型组相比,茵陈蒿汤组PPARα、CPT1A、CPT2、MCAD、LCAD、ACOX1、EHHADH mRNA和蛋白表达水平显著升高(P<0.05)。结论茵陈蒿汤可能通过调控Lnc MEG3/miR-21-5p/PPARα信号通路促进肝脏脂肪酸β氧化,从而达到防治MAFLD的作用。展开更多
目的:探讨免疫球蛋白A肾病(IgAN)患者血浆中微小核糖核酸-21-5p(miR-21-5p)表达与肾功能及肾间质纤维化(RIF)的关系。方法:选取2021年1月—2023年1月我院收治IgAN患者100例为IgAN组,根据慢性肾脏病分期分为1期组22例、2期组30例、3期组2...目的:探讨免疫球蛋白A肾病(IgAN)患者血浆中微小核糖核酸-21-5p(miR-21-5p)表达与肾功能及肾间质纤维化(RIF)的关系。方法:选取2021年1月—2023年1月我院收治IgAN患者100例为IgAN组,根据慢性肾脏病分期分为1期组22例、2期组30例、3期组26例、4期组14例、5期组8例,根据牛津分型分为T0组32例、T1组42例、T2组26例,另选取同期50名体检健康志愿者为对照组。检测血浆miR-21-5p、肾功能指标、RIF指标。分析IgAN患者血浆miR-21-5p表达与肾功能及RIF指标的相关性。结果:与对照组比较,IgAN组血浆miR-21-5p、24 h尿蛋白定量(24 h UPQ)、单核细胞趋化蛋白-1(MCP-1)、转化生长因子-β_(1)(TGF-β_(1))水平升高,估计肾小球滤过率(eGFR)降低(P<0.05)。随着慢性肾脏病分期及牛津分型的增加,血浆miR-21-5p、24 h UPQ、MCP-1、TGF-β_(1)水平依次升高,eGFR依次降低(P<0.05)。IgAN患者血浆miR-21-5p表达与eGFR呈负相关,与24 h UPQ、MCP-1、TGF-β_(1)水平呈正相关(P<0.05)。结论:IgAN患者血浆miR-21-5p高表达,与肾功能降低和RIF密切相关。展开更多
BACKGROUND Heart disease remains a leading cause of mortality worldwide,with existing treatments often failing to effectively restore damaged myocardium.Humaninduced pluripotent stem cells(hiPSCs)and their derivatives...BACKGROUND Heart disease remains a leading cause of mortality worldwide,with existing treatments often failing to effectively restore damaged myocardium.Humaninduced pluripotent stem cells(hiPSCs)and their derivatives offer promising therapeutic options;however,challenges such as low retention,engraftment issues,and tumorigenic risks hinder their clinical utility.Recent focus has shifted to exosomes(exos)-nanoscale vesicles that facilitate intercellular communication-as a safer and more versatile alternative.Understanding the specific mechanisms and comparative efficacy of exos from hiPSCs vs hiPSC-derived cardiomyocytes(hiPSC-CMs)is crucial for advancing cardiac repair therapies.AIM To evaluate and compare the therapeutic efficacy of exos secreted by hiPSCs and hiPSC-CMs in cardiac repair,and to elucidate the role of microRNA 21-5p(miR-21-5p)in the observed effects.METHODS We differentiated hiPSCs into CMs using small molecule methods and characterized the cells and their exos.RESULTS Our findings indicate that hiPSC-CMs and their exos enhanced cardiac function,reduced infarct size,and decreased myocardial fibrosis in a murine myocardial infarction model.Notably,hiPSC-CM exos outperformed hiPSC-CM cell therapy,showing improved ejection fraction and reduced apoptosis.We identified miR-21-5p,a microRNA in hiPSC-CM exos,as crucial for CM survival.Exos with miR-21-5p were absorbed by AC16 cells,suggesting a mechanism for their cytoprotective effects.CONCLUSION Overall,hiPSC-CM exos could serve as a potent therapeutic agent for myocardial repair,laying the groundwork for future research into exos as a treatment for ischemic heart disease.展开更多
Spermatogonial stem cells(SSCs)are the key to maintaining production of the sperms and healthy offsprings,and also treating breeding livestock's reproductive damage and infertility.micro RNAs act a decisive role i...Spermatogonial stem cells(SSCs)are the key to maintaining production of the sperms and healthy offsprings,and also treating breeding livestock's reproductive damage and infertility.micro RNAs act a decisive role in regulating gene expression in many cells and tissues,including in processes such as proliferation,self-renewal,differentiation,and apoptosis of stem cells.However,the miRNA mechanism in regulation of SSCs is still unclear.Here,high-throughput sequencing was used to identify specific miRNAs.We confirmed that miR-21-5p was concentrated in both goat and mouse SSCs,and enhanced the proliferation and antiapoptotic ability of SSCs.In vivo experiments have shown that miR-21-5p resisted the damage of the chemotherapy drug Busulfan to germ cells,ameliorated Busulfan-induced testicular dysfunction,and maintained spermatogenesis.Further RNA-seq and target gene prediction revealed that SPRY1 and FASLG are targets of miR-21-5p,thereby activating downstream signaling pathways such as MAPK/ERK,PI3K-AKT,and apoptosis.In summary,miR-21-5p is crucial for the self-renewal and maintenance of SSCs.This study provides new avenues for treating breeding livestock's reproductive damages,infertility,oligospermia,and other conditions.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.
文摘目的:探讨免疫球蛋白A肾病(IgAN)患者血浆中微小核糖核酸-21-5p(miR-21-5p)表达与肾功能及肾间质纤维化(RIF)的关系。方法:选取2021年1月—2023年1月我院收治IgAN患者100例为IgAN组,根据慢性肾脏病分期分为1期组22例、2期组30例、3期组26例、4期组14例、5期组8例,根据牛津分型分为T0组32例、T1组42例、T2组26例,另选取同期50名体检健康志愿者为对照组。检测血浆miR-21-5p、肾功能指标、RIF指标。分析IgAN患者血浆miR-21-5p表达与肾功能及RIF指标的相关性。结果:与对照组比较,IgAN组血浆miR-21-5p、24 h尿蛋白定量(24 h UPQ)、单核细胞趋化蛋白-1(MCP-1)、转化生长因子-β_(1)(TGF-β_(1))水平升高,估计肾小球滤过率(eGFR)降低(P<0.05)。随着慢性肾脏病分期及牛津分型的增加,血浆miR-21-5p、24 h UPQ、MCP-1、TGF-β_(1)水平依次升高,eGFR依次降低(P<0.05)。IgAN患者血浆miR-21-5p表达与eGFR呈负相关,与24 h UPQ、MCP-1、TGF-β_(1)水平呈正相关(P<0.05)。结论:IgAN患者血浆miR-21-5p高表达,与肾功能降低和RIF密切相关。
基金Supported by the Nonprofit Research Institutes Foundation of Fujian Province,China,No.2021R1012005 and No.2021R1012003.
文摘BACKGROUND Heart disease remains a leading cause of mortality worldwide,with existing treatments often failing to effectively restore damaged myocardium.Humaninduced pluripotent stem cells(hiPSCs)and their derivatives offer promising therapeutic options;however,challenges such as low retention,engraftment issues,and tumorigenic risks hinder their clinical utility.Recent focus has shifted to exosomes(exos)-nanoscale vesicles that facilitate intercellular communication-as a safer and more versatile alternative.Understanding the specific mechanisms and comparative efficacy of exos from hiPSCs vs hiPSC-derived cardiomyocytes(hiPSC-CMs)is crucial for advancing cardiac repair therapies.AIM To evaluate and compare the therapeutic efficacy of exos secreted by hiPSCs and hiPSC-CMs in cardiac repair,and to elucidate the role of microRNA 21-5p(miR-21-5p)in the observed effects.METHODS We differentiated hiPSCs into CMs using small molecule methods and characterized the cells and their exos.RESULTS Our findings indicate that hiPSC-CMs and their exos enhanced cardiac function,reduced infarct size,and decreased myocardial fibrosis in a murine myocardial infarction model.Notably,hiPSC-CM exos outperformed hiPSC-CM cell therapy,showing improved ejection fraction and reduced apoptosis.We identified miR-21-5p,a microRNA in hiPSC-CM exos,as crucial for CM survival.Exos with miR-21-5p were absorbed by AC16 cells,suggesting a mechanism for their cytoprotective effects.CONCLUSION Overall,hiPSC-CM exos could serve as a potent therapeutic agent for myocardial repair,laying the groundwork for future research into exos as a treatment for ischemic heart disease.
基金supported by the National Natural Science Foundation of China(32072806 and 32372970)the National Key Research and Development Program of China(2022YFD1302201 and 2023YFF1000904)+3 种基金the Program of Shaanxi Province Science and Technology Innovation Team,China(2019TD-036)the Major Projects of Natural Science Foundation of Inner Mongolia Autonomous Region,China(2020ZD10)the Inner Mongolia Autonomous Region Competition Leaders,China(2022JBGS0025)Key Technologies Demonstration of Animal Husbandry in Shaanxi Province,China(20221086 and 20230978)。
文摘Spermatogonial stem cells(SSCs)are the key to maintaining production of the sperms and healthy offsprings,and also treating breeding livestock's reproductive damage and infertility.micro RNAs act a decisive role in regulating gene expression in many cells and tissues,including in processes such as proliferation,self-renewal,differentiation,and apoptosis of stem cells.However,the miRNA mechanism in regulation of SSCs is still unclear.Here,high-throughput sequencing was used to identify specific miRNAs.We confirmed that miR-21-5p was concentrated in both goat and mouse SSCs,and enhanced the proliferation and antiapoptotic ability of SSCs.In vivo experiments have shown that miR-21-5p resisted the damage of the chemotherapy drug Busulfan to germ cells,ameliorated Busulfan-induced testicular dysfunction,and maintained spermatogenesis.Further RNA-seq and target gene prediction revealed that SPRY1 and FASLG are targets of miR-21-5p,thereby activating downstream signaling pathways such as MAPK/ERK,PI3K-AKT,and apoptosis.In summary,miR-21-5p is crucial for the self-renewal and maintenance of SSCs.This study provides new avenues for treating breeding livestock's reproductive damages,infertility,oligospermia,and other conditions.
