OBJECTIVE:To evaluate the protective effects of Chang'an decoction(肠安方,CAD)on colitis,and investigate the potential mechanisms underlying these effects from the perspectives of endoplasmic reticulum(ER)stress i...OBJECTIVE:To evaluate the protective effects of Chang'an decoction(肠安方,CAD)on colitis,and investigate the potential mechanisms underlying these effects from the perspectives of endoplasmic reticulum(ER)stress induced by mitofusin 2(MFN2).METHODS:The composition of CAD was identified by liquid chromatography-mass spectrometry technology.A mice model of dextran sulfate sodium(DSS)induced colitis was established and therapeutic effects of CAD were determined by detecting body weight,disease activity index,colon length and histopathological changes.Then,the expression levels of MFN2,ER stress markers and Nucleotide-binding domain and leucine-rich repeat protein3(NLRP3)relevant proteins were detected by polymerase chain reaction(PCR),Western blot,immunohistochemistry and immunofluorescence staining.Subsequently,knockdown and overexpression cell model were constructed to further investigate the underlying mechanism of MFN2 mediating ER stress and energy metabolism by PCR,Western blot,electron microscopy and reactive oxygen species(ROS)staining.Finally,inflammatory indicator and tight junction proteins were measured by PCR and immunofluorescence staining to evaluate the protective effects of CAD.RESULTS:Results showed that the indispensable regulatory role of MFN2 in mediating ER stress and mitochondrial damage was involved in the protective effects of CAD on colitis in mice fed with DSS.Network pharmacology analysis also revealed CAD may play a protective effect on colitis by affecting mitochondrial function.In addition,our data also suggested a causative role for MFN2 in the development of inflammatory responses and energy metabolic alterations by constructing a knockdown and overexpression cell model whereby alter proper ER-mitochondria interaction in Caco-2 cells.Furthermore,relative expression analyses of ER stress markers and NLRP3 inflammasome showed the onset of ER stress and activation of NLRP3 inflammasome,which is consistent with the above findings.In contrast,intervention of CAD could improve the mucosal barrier integrity and colonic inflammatory response effectively through inhibiting ER stress response mediated by MFN2.CONCLUSION:CAD could alleviate ER stress by regulating MFN2 to exert therapeutic effects on DSS-induced colitis,which might provide an effective natural therapeutic approach for the treatment of ulcerative colitis.展开更多
基金the National Natural Science Foundation of China Youth Fund:Study on the Mechanism of Chang'an Decoction Regulating the Endoplasmic Reticulum Stress Response via Mfn2 in the Treatment of Ulcerative Colitis(No.82004305)and the National Natural Science Foundation of China Youth Fund:Study on Protective Mechanisms of Longqi Jiangzhi Decoction against Non-alcoholic Steatohepatitis based on Th17 Cells Differentiation Induced by SPP1(No.82104549)。
文摘OBJECTIVE:To evaluate the protective effects of Chang'an decoction(肠安方,CAD)on colitis,and investigate the potential mechanisms underlying these effects from the perspectives of endoplasmic reticulum(ER)stress induced by mitofusin 2(MFN2).METHODS:The composition of CAD was identified by liquid chromatography-mass spectrometry technology.A mice model of dextran sulfate sodium(DSS)induced colitis was established and therapeutic effects of CAD were determined by detecting body weight,disease activity index,colon length and histopathological changes.Then,the expression levels of MFN2,ER stress markers and Nucleotide-binding domain and leucine-rich repeat protein3(NLRP3)relevant proteins were detected by polymerase chain reaction(PCR),Western blot,immunohistochemistry and immunofluorescence staining.Subsequently,knockdown and overexpression cell model were constructed to further investigate the underlying mechanism of MFN2 mediating ER stress and energy metabolism by PCR,Western blot,electron microscopy and reactive oxygen species(ROS)staining.Finally,inflammatory indicator and tight junction proteins were measured by PCR and immunofluorescence staining to evaluate the protective effects of CAD.RESULTS:Results showed that the indispensable regulatory role of MFN2 in mediating ER stress and mitochondrial damage was involved in the protective effects of CAD on colitis in mice fed with DSS.Network pharmacology analysis also revealed CAD may play a protective effect on colitis by affecting mitochondrial function.In addition,our data also suggested a causative role for MFN2 in the development of inflammatory responses and energy metabolic alterations by constructing a knockdown and overexpression cell model whereby alter proper ER-mitochondria interaction in Caco-2 cells.Furthermore,relative expression analyses of ER stress markers and NLRP3 inflammasome showed the onset of ER stress and activation of NLRP3 inflammasome,which is consistent with the above findings.In contrast,intervention of CAD could improve the mucosal barrier integrity and colonic inflammatory response effectively through inhibiting ER stress response mediated by MFN2.CONCLUSION:CAD could alleviate ER stress by regulating MFN2 to exert therapeutic effects on DSS-induced colitis,which might provide an effective natural therapeutic approach for the treatment of ulcerative colitis.
