Viral myocarditis(VMC)is one of the most common acquired heart diseases in children and teenagers.However,its pathogenesis is still unclear,and effective treatments are lacking.This study aimed to investigate the regu...Viral myocarditis(VMC)is one of the most common acquired heart diseases in children and teenagers.However,its pathogenesis is still unclear,and effective treatments are lacking.This study aimed to investigate the regulatory pathway by which exosomes alleviate ferroptosis in cardiomyocytes(CMCs)induced by coxsackievirus B3(CVB3).CVB3 was utilized for inducing the VMC mouse model and cellular model.Cardiac echocardiography,left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS)were implemented to assess the cardiac function.In CVB3-induced VMC mice,cardiac insufficiency was observed,as well as the altered levels of ferroptosis-related indicators(glutathione) peroxidase 4(GPX4),glutathione(GSH),and malondialdehyde(MDA).However,exosomes derived from human umbilical cord mesenchymal stem cells(hucMSCs-exo)could restore the changes caused by CVB3 stimulation.Let-7a-5p was enriched in hucMSCs-exo,and the inhibitory ffect of hucMSCs-exoa-ie-pmimo on CVB3-induced ferroptosis was higher than that of hucMSCs-exommie N(NC:negative control).Mothers against decapentaplegic homolog 2(SMAD2)increased in the VMC group,while the expression of zinc-finger protein 36(ZFP36)decreased.Let-7a-5p was confirmed to interact with SMAD2 messenger RNA(mRNA),and the SMAD2 protein interacted directly with the ZFP36 protein.Silencing SMAD2 and overexpressing ZFP36 inhibited the expression of ferroptosis-related indicators.Meanwhile,the levels of GPX4,solute carrier family 7,member 11(SLC7A11),and GSH were lower in the SMAD2 overexpression plasmid(oe-SMAD2)+let-7a-5p mimic group than in the oe-NC+let-7a-5p mimic group,while those of MDA,reactive oxygen species(ROS),and Fe^(2+)increased.In conclusion,these data showed that ferroptosis could be regulated by mediating SMAD2 expression.Exo-let-7a-5p derived from hucMSCs could mediate SMAD2 to promote the expression of ZFP36,which further inhibited the ferroptosis of CMCs to alleviate CVB3-induced VMC.展开更多
Long noncoding RNAs(lncRNAs)are expressed in different species and different tissues,and perform different functions,but little is known about their involvement in the synthesis or secretion of follicle-stimulating ho...Long noncoding RNAs(lncRNAs)are expressed in different species and different tissues,and perform different functions,but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone(FSH).In general,we have revealed lnc RNA-micro RNA(mi RNA)-messenger RNA(m RNA)interactions that may play important roles in rat primary pituitary cells.In this study,a new lncRNA was identified for the first time.First,we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization,which indicated that this lncRNA was distributed mainly in the cytoplasm.Next,we used RT-qPCR and enzyme-linked immunosorbent assay(ELISA)to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion.In addition,mothers against decapentaplegic homolog 2(Smad2)was highly expressed in our sequencing results.We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2.We used RNA immunoprecipitation-qPCR(RIP-qPCR)and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2.Fluorescence in situ hybridization(FISH)showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm.Finally,we determined the relationship among lncRNA-m18as1,miR-18a-5p,and the Smad2/3 pathway.Overall,we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.展开更多
目的研究中药颗粒对染矽尘、Col-Ⅰ的影响,并从TGF-β1/Smads通路探讨其可能的作用机制。方法将SD雄性大鼠随机分为对照组、模型组和中药高、低剂量组,每组16只。对照组大鼠气管灌注1.0 ml生理盐水,其余组大鼠均给予1.0 ml 5%无菌SiO2...目的研究中药颗粒对染矽尘、Col-Ⅰ的影响,并从TGF-β1/Smads通路探讨其可能的作用机制。方法将SD雄性大鼠随机分为对照组、模型组和中药高、低剂量组,每组16只。对照组大鼠气管灌注1.0 ml生理盐水,其余组大鼠均给予1.0 ml 5%无菌SiO2悬浊液;造模后第2天开始给大鼠灌胃给药,1次/d,高剂量组、低剂量组大鼠分别灌胃1 000 mg/kg、500 mg/kg中药,对照组、模型组给予0.9%生理盐水。于灌胃第14、28天处死大鼠,并检测大鼠肺组织Col-Ⅰ、TGF-β1的含量及TGF-β1、Smad2/3蛋白的表达水平。结果造模后第14天和第28天,模型组大鼠肺组织Col-Ⅰ、TGF-β1含量高于对照组,中药干预后Col-Ⅰ、TGF-β1含量有所降低,不同剂量组间差异均具有统计学意义(P<0.05);模型组大鼠肺组织TGF-β1、Smad2/3及p-Smad2/3蛋白的表达水平高于对照组,中药干预后TGF-β1、Smad2/3及p-Smad2/3蛋白的表达水平有所降低,不同剂量组间差异也具有统计学意义(P<0.05)。结论中药颗粒可以抑制SiO2诱导的大鼠肺组织Col-Ⅰ含量的升高,且该抑制作用可能与中药颗粒对大鼠肺组织TGF-β1/Smads通路的调节有关。展开更多
基金supported by the China Postdoctoral Science Foundation(No.2022M712252)the Natural Science Foundation of Sichuan Province,China(No.2023NSFSC1634).
文摘Viral myocarditis(VMC)is one of the most common acquired heart diseases in children and teenagers.However,its pathogenesis is still unclear,and effective treatments are lacking.This study aimed to investigate the regulatory pathway by which exosomes alleviate ferroptosis in cardiomyocytes(CMCs)induced by coxsackievirus B3(CVB3).CVB3 was utilized for inducing the VMC mouse model and cellular model.Cardiac echocardiography,left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS)were implemented to assess the cardiac function.In CVB3-induced VMC mice,cardiac insufficiency was observed,as well as the altered levels of ferroptosis-related indicators(glutathione) peroxidase 4(GPX4),glutathione(GSH),and malondialdehyde(MDA).However,exosomes derived from human umbilical cord mesenchymal stem cells(hucMSCs-exo)could restore the changes caused by CVB3 stimulation.Let-7a-5p was enriched in hucMSCs-exo,and the inhibitory ffect of hucMSCs-exoa-ie-pmimo on CVB3-induced ferroptosis was higher than that of hucMSCs-exommie N(NC:negative control).Mothers against decapentaplegic homolog 2(SMAD2)increased in the VMC group,while the expression of zinc-finger protein 36(ZFP36)decreased.Let-7a-5p was confirmed to interact with SMAD2 messenger RNA(mRNA),and the SMAD2 protein interacted directly with the ZFP36 protein.Silencing SMAD2 and overexpressing ZFP36 inhibited the expression of ferroptosis-related indicators.Meanwhile,the levels of GPX4,solute carrier family 7,member 11(SLC7A11),and GSH were lower in the SMAD2 overexpression plasmid(oe-SMAD2)+let-7a-5p mimic group than in the oe-NC+let-7a-5p mimic group,while those of MDA,reactive oxygen species(ROS),and Fe^(2+)increased.In conclusion,these data showed that ferroptosis could be regulated by mediating SMAD2 expression.Exo-let-7a-5p derived from hucMSCs could mediate SMAD2 to promote the expression of ZFP36,which further inhibited the ferroptosis of CMCs to alleviate CVB3-induced VMC.
基金the National Natural Science Foundation of China(No.31872349)。
文摘Long noncoding RNAs(lncRNAs)are expressed in different species and different tissues,and perform different functions,but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone(FSH).In general,we have revealed lnc RNA-micro RNA(mi RNA)-messenger RNA(m RNA)interactions that may play important roles in rat primary pituitary cells.In this study,a new lncRNA was identified for the first time.First,we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction(RT-qPCR)and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization,which indicated that this lncRNA was distributed mainly in the cytoplasm.Next,we used RT-qPCR and enzyme-linked immunosorbent assay(ELISA)to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion.In addition,mothers against decapentaplegic homolog 2(Smad2)was highly expressed in our sequencing results.We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2.We used RNA immunoprecipitation-qPCR(RIP-qPCR)and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2.Fluorescence in situ hybridization(FISH)showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm.Finally,we determined the relationship among lncRNA-m18as1,miR-18a-5p,and the Smad2/3 pathway.Overall,we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.