The objective of this study was to determine the role of SLC15A4 in the muramyl dipeptide(MDP)-mediated inflammatory response of bovine rumen epithelial cells(BRECs).First,changes in the m RNA expression of proinflamm...The objective of this study was to determine the role of SLC15A4 in the muramyl dipeptide(MDP)-mediated inflammatory response of bovine rumen epithelial cells(BRECs).First,changes in the m RNA expression of proinflammatory factor genes in BRECs following 10μg m L^(–1)MDP treatments were examined.RT-q PCR results showed that the expression of pro-inflammatory factor(IL-1β,IL-6,and TNF-α)m RNAs were significantly increased under MDP stimulation(P<0.001).Moreover,SLC15A4-Knockout(SLC15A4-KO)cells were obtained through lentivirus packaging,transfection,screening,and cell monoclonal culture.In order to gain further insight into the potential function of SLC15A4,we utilized transcriptome data,which revealed a change in the genes between WT-BRECs and SLC15A4-KO.Five down-regulated pro-inflammatory genes and 13 down-regulated chemokine genes related to the inflammatory response were identified.Meanwhile,the down-regulated genes were mostly enriched in the nuclear factorκB(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways.The results of RT-q PCR also verified these detected changes.To further determine the mechanism of how WT and SLC15A4-KO BRECs are involved in inflammatory responses,we investigated the inflammatory responses of cells exposed to MDP.WT-BRECs and SLC15A4-KO were treated with a culture medium containing 10μg m L^(–1)MDP,in comparison to a control without MDP.Our results show that SLC15A4-KO BRECs had reduced the expression of genes(IL-6,TNF-α,CXCL2,CXCL3,CXCL9,and CCL2)and proteins(p-p65 and p-p44/42)from the MDP-mediated inflammatory response compared to WT-BRECs(P<0.05).In this experiment,CRISPR-Cas9 was used to KO the di/tripeptide transporter SLC15A4,and its role was confirmed via the MDP-induced inflammatory response in BRECs.This work will provide a theoretical basis for studying the pro-inflammatory mechanism of MDP and its application in the prevention and treatment of subacute rumen acidosis in dairy cows.展开更多
伴随信息产业的快速发展和互联网保险的广泛运用,近几年来国际上出现了一种新型保险模式――P2P保险(Peer to Peer Insurance)。该模式基于互联网,通过聚集若干风险类型相似的投保人组建风险共担互助小组,风险互助小组成员间一般是亲戚...伴随信息产业的快速发展和互联网保险的广泛运用,近几年来国际上出现了一种新型保险模式――P2P保险(Peer to Peer Insurance)。该模式基于互联网,通过聚集若干风险类型相似的投保人组建风险共担互助小组,风险互助小组成员间一般是亲戚、朋友等熟人关系。P2P保险形成的相互监督机制和声誉机制,可以较好地解决传统保险市场中存在的道德风险问题。本文利用经济学中的比较静态分析方法,研究了P2P保险的道德风险问题,并从理论上证明了在P2P保险模式下,投保人会倾向于更加努力防范风险以降低出险概率,道德风险问题可以得到有效缓解。展开更多
基金the National Natural Science Foundation of China(31972589)the earmarked fund for China Agriculture Research System(CARS-36)the Postgraduate Research&Practice Innovation Program of Jiangsu Province,China(KYCX21-3283)。
文摘The objective of this study was to determine the role of SLC15A4 in the muramyl dipeptide(MDP)-mediated inflammatory response of bovine rumen epithelial cells(BRECs).First,changes in the m RNA expression of proinflammatory factor genes in BRECs following 10μg m L^(–1)MDP treatments were examined.RT-q PCR results showed that the expression of pro-inflammatory factor(IL-1β,IL-6,and TNF-α)m RNAs were significantly increased under MDP stimulation(P<0.001).Moreover,SLC15A4-Knockout(SLC15A4-KO)cells were obtained through lentivirus packaging,transfection,screening,and cell monoclonal culture.In order to gain further insight into the potential function of SLC15A4,we utilized transcriptome data,which revealed a change in the genes between WT-BRECs and SLC15A4-KO.Five down-regulated pro-inflammatory genes and 13 down-regulated chemokine genes related to the inflammatory response were identified.Meanwhile,the down-regulated genes were mostly enriched in the nuclear factorκB(NF-κB)and mitogen-activated protein kinase(MAPK)signaling pathways.The results of RT-q PCR also verified these detected changes.To further determine the mechanism of how WT and SLC15A4-KO BRECs are involved in inflammatory responses,we investigated the inflammatory responses of cells exposed to MDP.WT-BRECs and SLC15A4-KO were treated with a culture medium containing 10μg m L^(–1)MDP,in comparison to a control without MDP.Our results show that SLC15A4-KO BRECs had reduced the expression of genes(IL-6,TNF-α,CXCL2,CXCL3,CXCL9,and CCL2)and proteins(p-p65 and p-p44/42)from the MDP-mediated inflammatory response compared to WT-BRECs(P<0.05).In this experiment,CRISPR-Cas9 was used to KO the di/tripeptide transporter SLC15A4,and its role was confirmed via the MDP-induced inflammatory response in BRECs.This work will provide a theoretical basis for studying the pro-inflammatory mechanism of MDP and its application in the prevention and treatment of subacute rumen acidosis in dairy cows.
文摘伴随信息产业的快速发展和互联网保险的广泛运用,近几年来国际上出现了一种新型保险模式――P2P保险(Peer to Peer Insurance)。该模式基于互联网,通过聚集若干风险类型相似的投保人组建风险共担互助小组,风险互助小组成员间一般是亲戚、朋友等熟人关系。P2P保险形成的相互监督机制和声誉机制,可以较好地解决传统保险市场中存在的道德风险问题。本文利用经济学中的比较静态分析方法,研究了P2P保险的道德风险问题,并从理论上证明了在P2P保险模式下,投保人会倾向于更加努力防范风险以降低出险概率,道德风险问题可以得到有效缓解。
