Background:Inflammation plays an important role in the pathogenesis of status epilepticus(SE).The long non-cod-ing RNA(lncRNA)taurine up-regulated gene1(Tug1)plays a well-defned role in inflammatory diseases.However,t...Background:Inflammation plays an important role in the pathogenesis of status epilepticus(SE).The long non-cod-ing RNA(lncRNA)taurine up-regulated gene1(Tug1)plays a well-defned role in inflammatory diseases.However,the molecular mechanism of Tug1 in SE progression remains unknown.In present study,we investigated whether Tug1 is involved in microglial inflammation in SE rats.Methods:The SE rat model was established via intraperitoneal injection of lithium chloride pilocarpine.RNA-binding protein immunoprecipitation(RIP)and RIP sequencing were carried out in rat microglia(RM).Tug1 cloned into the adenovirus was overexpressed in the microglia.Knockdown of Tug1 was performed via siRNA transfection.The level of Tug1 and inflammatory factors IL-1βand TNF-αwas examined by real-time polymerase chain reaction(RT-PCR)and western blotting.Protein levels of p65,P-p65,p-IKBa and IkBa were assessed by western blotting.Results:The RIP-seq result showed 14 lncRNAs that bound to the NF-κB p65 protein in RM.The lncRNA Tug1 directly interacted with p65.The level of declined Tug1 was decreased in the hippocampus of SE rats.Overexpression ofTug1 reduced the LPS-induced inflammation and M1/M2 polarization of microglia,while knockdown of Tug1 aggravated the inflammatory response in microglia.Accordingly,the protein levels of p-p65/p65 and p-IkBa/IkBa were reduced in the Tug1-overexpression microglia and elevated in the Tug1-knockdown microglia.Conclusions:These findings indicate that Tug1 modulates the inflammation in microglia through the NF-κB signal pathway,and the Tug1/P65 axis are like to play a signifcant role in the inflammatory processes,providing a valid target for the therapy of SE.展开更多
Skeletal muscle differentiation is a highly coordinated process that involves many cellular signaling pathways and microRNAs(miRNAs).A group of muscle-specific miRNAs has been reported to promote myogenesis by suppres...Skeletal muscle differentiation is a highly coordinated process that involves many cellular signaling pathways and microRNAs(miRNAs).A group of muscle-specific miRNAs has been reported to promote myogenesis by suppressing key signaling pathways for cell growth.However,the functional role and regulatory mechanism of most non-muscle-specific miRNAs with stage-specific changes during differentiation are largely unclear.Here,we describe the functional characterization of miR-101a/b,a pair of non-muscle-specific miRNAs that show the largest change among a group of transiently upregulated miRNAs during myogenesis in C2C12 cells.The overexpression of miR-101a/b inhibits myoblast differentiation by suppressing the p38/MAPK,Interferon Gamma,and Wnt pathways and enhancing the C/EBP pathway.Mef2a,a key protein in the p38/MAPK pathway,was identified as a direct target of miR-101a/b.Interestingly,we found that the long non-coding RNA(lncRNA)Malat1,which promotes muscle differentiation,interacts with miR-101a/b,and this interaction competes with Mef2a mRNA to relieve the inhibition of the p38/MAPK pathway during myogenesis.These results uncovered a“braking”role in differentiation of transiently upregulated miRNAs and provided new insights into the competing endogenous RNA(ceRNA)regulatory mechanism in myoblast differentiation and myogenesis.展开更多
Long noncoding RNAs(lncRNAs)modulate many aspects of biological and pathological processes.Recent studies have shown that host lncRNAs participate in the antiviral immune response,but functional lncRNAs in coxsackievi...Long noncoding RNAs(lncRNAs)modulate many aspects of biological and pathological processes.Recent studies have shown that host lncRNAs participate in the antiviral immune response,but functional lncRNAs in coxsackievirus B5(CVB5)infection remain unknown.Here,we identified a novel cytoplasmic lncRNA,LINC1392,which was highly inducible in CVB5 infected RD cells in a time-and dose-dependent manner,and also can be induced by the viral RNA and IFN-β.Further investigation showed that LINC1392 promoted several important interferon-stimulated genes(ISGs)expression,including IFIT1,IFIT2,and IFITM3 by activating MDA5,thereby inhibiting the replication of CVB5 in vitro.Mechanistically,LINC1392 bound to ELAV like RNA binding protein 1(ELAVL1)and blocked ELAVL1 interaction with MDA5.Functional study revealed that the 245–835 nt locus of LINC1392 exerted the antiviral effect and was also an important site for ELAVL1 binding.In mice,LINC1392 could inhibit CVB5 replication and alleviated the histopathological lesions of intestinal and brain tissues induced by viral infection.Our findings collectively reveal that the novel LINC1392 acts as a positive regulator in the IFN-I signaling pathway against CVB5 infection.Elucidating the underlying mechanisms on how lncRNA regulats the host innate immunity response towards CVB5 infection will lay the foundation for antiviral drug research.展开更多
基金the Ethics Committee of Shanghai public clinical health center of Fudan University(No.2019JS015)。
文摘Background:Inflammation plays an important role in the pathogenesis of status epilepticus(SE).The long non-cod-ing RNA(lncRNA)taurine up-regulated gene1(Tug1)plays a well-defned role in inflammatory diseases.However,the molecular mechanism of Tug1 in SE progression remains unknown.In present study,we investigated whether Tug1 is involved in microglial inflammation in SE rats.Methods:The SE rat model was established via intraperitoneal injection of lithium chloride pilocarpine.RNA-binding protein immunoprecipitation(RIP)and RIP sequencing were carried out in rat microglia(RM).Tug1 cloned into the adenovirus was overexpressed in the microglia.Knockdown of Tug1 was performed via siRNA transfection.The level of Tug1 and inflammatory factors IL-1βand TNF-αwas examined by real-time polymerase chain reaction(RT-PCR)and western blotting.Protein levels of p65,P-p65,p-IKBa and IkBa were assessed by western blotting.Results:The RIP-seq result showed 14 lncRNAs that bound to the NF-κB p65 protein in RM.The lncRNA Tug1 directly interacted with p65.The level of declined Tug1 was decreased in the hippocampus of SE rats.Overexpression ofTug1 reduced the LPS-induced inflammation and M1/M2 polarization of microglia,while knockdown of Tug1 aggravated the inflammatory response in microglia.Accordingly,the protein levels of p-p65/p65 and p-IkBa/IkBa were reduced in the Tug1-overexpression microglia and elevated in the Tug1-knockdown microglia.Conclusions:These findings indicate that Tug1 modulates the inflammation in microglia through the NF-κB signal pathway,and the Tug1/P65 axis are like to play a signifcant role in the inflammatory processes,providing a valid target for the therapy of SE.
基金supported by the National Natural Science Foundation of China(31970604,31701116,31770879,31771459,31900903,81870449,81974436)the Major Research Plan of the National Natural Science Foundation of China(91940000)+1 种基金the Fundamental Research Funds for the Central Universities(20lgpy112)Science and Technology New Star in ZhuJiang Guangzhou City(201806010151).
文摘Skeletal muscle differentiation is a highly coordinated process that involves many cellular signaling pathways and microRNAs(miRNAs).A group of muscle-specific miRNAs has been reported to promote myogenesis by suppressing key signaling pathways for cell growth.However,the functional role and regulatory mechanism of most non-muscle-specific miRNAs with stage-specific changes during differentiation are largely unclear.Here,we describe the functional characterization of miR-101a/b,a pair of non-muscle-specific miRNAs that show the largest change among a group of transiently upregulated miRNAs during myogenesis in C2C12 cells.The overexpression of miR-101a/b inhibits myoblast differentiation by suppressing the p38/MAPK,Interferon Gamma,and Wnt pathways and enhancing the C/EBP pathway.Mef2a,a key protein in the p38/MAPK pathway,was identified as a direct target of miR-101a/b.Interestingly,we found that the long non-coding RNA(lncRNA)Malat1,which promotes muscle differentiation,interacts with miR-101a/b,and this interaction competes with Mef2a mRNA to relieve the inhibition of the p38/MAPK pathway during myogenesis.These results uncovered a“braking”role in differentiation of transiently upregulated miRNAs and provided new insights into the competing endogenous RNA(ceRNA)regulatory mechanism in myoblast differentiation and myogenesis.
基金This work was supported by the National Natural Science Foundation of China(No.81860357)the Young Talents Support Program of Yunnan Province,China(Ten Thousand People Plan,YNWR-QNBJ-2019-178).
文摘Long noncoding RNAs(lncRNAs)modulate many aspects of biological and pathological processes.Recent studies have shown that host lncRNAs participate in the antiviral immune response,but functional lncRNAs in coxsackievirus B5(CVB5)infection remain unknown.Here,we identified a novel cytoplasmic lncRNA,LINC1392,which was highly inducible in CVB5 infected RD cells in a time-and dose-dependent manner,and also can be induced by the viral RNA and IFN-β.Further investigation showed that LINC1392 promoted several important interferon-stimulated genes(ISGs)expression,including IFIT1,IFIT2,and IFITM3 by activating MDA5,thereby inhibiting the replication of CVB5 in vitro.Mechanistically,LINC1392 bound to ELAV like RNA binding protein 1(ELAVL1)and blocked ELAVL1 interaction with MDA5.Functional study revealed that the 245–835 nt locus of LINC1392 exerted the antiviral effect and was also an important site for ELAVL1 binding.In mice,LINC1392 could inhibit CVB5 replication and alleviated the histopathological lesions of intestinal and brain tissues induced by viral infection.Our findings collectively reveal that the novel LINC1392 acts as a positive regulator in the IFN-I signaling pathway against CVB5 infection.Elucidating the underlying mechanisms on how lncRNA regulats the host innate immunity response towards CVB5 infection will lay the foundation for antiviral drug research.
