OBJECTIVE Atherosclerosis(AS)is a chronic inflammatory disease characterized by the accumulation of lipids,vascular fibrosis,and inflammation.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chi...OBJECTIVE Atherosclerosis(AS)is a chronic inflammatory disease characterized by the accumulation of lipids,vascular fibrosis,and inflammation.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits anti-AS effects.Our previous work demonstrated that gut microbiota plays an important role during AS treatment as it affects the efficacy of Pae.However,the mechanism of Pae in protecting against vascular fibrosis as related to gut microbiota has yet to be elucidated.To investigate the anti-fibrosis effect of Pae on AS mice and demonstrate the underlying gut microbiota-dependent mechanism.METHODS ApoE-/-mice were fed with high-fat-diet(HFD)to replicate the AS model.HE and Masson staining were used to observe the plaque formation and collagen deposition.Gut microbiota alteration and short-chain fatty acids(SCFAs)production were analyzed through 16S rRNA sequencing and LC-MS/MS.The frequency of immune cells in spleen were phenotyped by flow cytometry.The mRNA expression of aortic inflammatory cytokines were detected by qRT-PCR.The protein expression of LOX and fibrosis related indicators were examined by Western blotting.RESULTS Pae restricted the development of AS and collagen deposition.Notably,the anti-fibrosis effect of Pae was achieved by regulating the gut microbiota.16S rRNA sequencing and LC-MS/MS data indicated that the relative abundance of SCFAs-producing bacteria and SCFAs production was increased.Additionally,Pae administration selectively up-regulated the frequency of regulatory T(Treg)cells as well as down-regulated the ratio of T helper type 17(Th17)cells in the spleen of AS mice,improving the Treg/Th17 balance.In addition,as expected,Pae intervention significantly down-regulate the mRNA expression levels of pro-inflammatory cytokines IL^(-1)β,IL-6,TNF-αand IL^(-1)7 in the aorta tissue,up-regulate the levels of anti-inflammatory factor IL^(-1)0,a marker of Treg cells.Finally,Pae′s intervention in the gut microbiota resulted in the restoration of the balance of Treg/Th17,which indirectly down-regulated the protein expression level of LOX and fibrosis-related indicators(MMP-2/9 and collagenⅠ/Ⅲ).CONCLUSION Pae attenuates vascular fibrosis in a gut microbiota-dependent manner.The underlying protective mechanism is associated with the improved Treg/Th17 balance in spleen mediated through the increased microbiota-derived SCFAs production.展开更多
OBJECTIVE Atherosclerosis(AS)is featured as a chronic inflammatory disease of vascular stenosis.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits ant...OBJECTIVE Atherosclerosis(AS)is featured as a chronic inflammatory disease of vascular stenosis.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits anti-AS effects in vitro and in vivo.In this study,we aimed to investigate whether the anti-AS efficacy of Pae was regulated through inhibiting NLRP3 inflammasome activityvia elevating hyperlipidemic rats plasma exosomalmicroR⁃NA-223(miR-223).METHODS The Sprague-Dawley rats was induced by a high-fat diet,which was used as AS models.AS aortic pathological morphological in AS mice was examined by HE staining,and serum TC and TG levels were deter⁃mined by automatic chemistry analyzer.Rat aortic endothelial cells(RAECs)were used during the whole study.After oral administration of Pae,we isolated exosomes from hyperlipidemic rats plasma(Pae-Exos)by ultracentrifugation and characterized by transmission electron,nanoparticle tracking analysis,dynamic light scattering and Western blotting.The activity of RAECs was detected by CCK-8 and trypan blue staining method.IL-1βand IL-6 levels were detected by ELISA method.The expression of miR-223 was detected by qPCR,and the expression of NLRP3,ASC,caspase-1,and ICAM-1 was detected by Western blotting.RESULTS In vivo experiments confirmed that Pae could effectively reduce serum TC and TG levels and decrease serum IL-1βand IL-6 levels,which demonstrated that Pae restricted AS develop⁃ment in hyperlipidemia rats.Both CCK-8 and trypan blue staining showed that the survival rate of RAECs in the Pae-Exos co-incubation group was higher than that in the model group.We also confirmed via real-time qPCR that Pae-Exos suppressed the expression of the inflammatory cytokines IL-1βand IL-6.Accordingly,Pae-Exos dose-dependently increased the survival rate of RAECs and reduced inflammatory response.Furthermore,compared with the model group,Pae-Exos more successfully increased the expression of miR-223 and inhibited IL-1βand IL-6 expression,which implied that Pae-exo may inhibited the inflammatory response of RAECs by increasing the content of miR-223.Subse⁃quently,we found that Pae-Exos reduced the expressions of NLRP3,ASC,caspase-1 and ICAM-1,which indicated that Pae-Exos may reduced RAECs inflammation by suppressing NLRP3 signaling pathway via promoting miR-223 expression.CONCLUSION Pae can inhibit the downstream NLRP3 inflammatory corpuscle signaling pathway by increasing the level of miR-223 in plasma Exos of hyperlipidemic rats,providing new insights into the anti-atherosclerosis activity of Pae.展开更多
基金National Natural Science Foundation of China(81773937)。
文摘OBJECTIVE Atherosclerosis(AS)is a chronic inflammatory disease characterized by the accumulation of lipids,vascular fibrosis,and inflammation.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits anti-AS effects.Our previous work demonstrated that gut microbiota plays an important role during AS treatment as it affects the efficacy of Pae.However,the mechanism of Pae in protecting against vascular fibrosis as related to gut microbiota has yet to be elucidated.To investigate the anti-fibrosis effect of Pae on AS mice and demonstrate the underlying gut microbiota-dependent mechanism.METHODS ApoE-/-mice were fed with high-fat-diet(HFD)to replicate the AS model.HE and Masson staining were used to observe the plaque formation and collagen deposition.Gut microbiota alteration and short-chain fatty acids(SCFAs)production were analyzed through 16S rRNA sequencing and LC-MS/MS.The frequency of immune cells in spleen were phenotyped by flow cytometry.The mRNA expression of aortic inflammatory cytokines were detected by qRT-PCR.The protein expression of LOX and fibrosis related indicators were examined by Western blotting.RESULTS Pae restricted the development of AS and collagen deposition.Notably,the anti-fibrosis effect of Pae was achieved by regulating the gut microbiota.16S rRNA sequencing and LC-MS/MS data indicated that the relative abundance of SCFAs-producing bacteria and SCFAs production was increased.Additionally,Pae administration selectively up-regulated the frequency of regulatory T(Treg)cells as well as down-regulated the ratio of T helper type 17(Th17)cells in the spleen of AS mice,improving the Treg/Th17 balance.In addition,as expected,Pae intervention significantly down-regulate the mRNA expression levels of pro-inflammatory cytokines IL^(-1)β,IL-6,TNF-αand IL^(-1)7 in the aorta tissue,up-regulate the levels of anti-inflammatory factor IL^(-1)0,a marker of Treg cells.Finally,Pae′s intervention in the gut microbiota resulted in the restoration of the balance of Treg/Th17,which indirectly down-regulated the protein expression level of LOX and fibrosis-related indicators(MMP-2/9 and collagenⅠ/Ⅲ).CONCLUSION Pae attenuates vascular fibrosis in a gut microbiota-dependent manner.The underlying protective mechanism is associated with the improved Treg/Th17 balance in spleen mediated through the increased microbiota-derived SCFAs production.
基金National Natural Science Foundation of China(8147338681773937)
文摘OBJECTIVE Atherosclerosis(AS)is featured as a chronic inflammatory disease of vascular stenosis.Paeonol(Pae)is a natural phenolic compounds isolated from a traditional Chinese medicine,Cortex Moutan,which exhibits anti-AS effects in vitro and in vivo.In this study,we aimed to investigate whether the anti-AS efficacy of Pae was regulated through inhibiting NLRP3 inflammasome activityvia elevating hyperlipidemic rats plasma exosomalmicroR⁃NA-223(miR-223).METHODS The Sprague-Dawley rats was induced by a high-fat diet,which was used as AS models.AS aortic pathological morphological in AS mice was examined by HE staining,and serum TC and TG levels were deter⁃mined by automatic chemistry analyzer.Rat aortic endothelial cells(RAECs)were used during the whole study.After oral administration of Pae,we isolated exosomes from hyperlipidemic rats plasma(Pae-Exos)by ultracentrifugation and characterized by transmission electron,nanoparticle tracking analysis,dynamic light scattering and Western blotting.The activity of RAECs was detected by CCK-8 and trypan blue staining method.IL-1βand IL-6 levels were detected by ELISA method.The expression of miR-223 was detected by qPCR,and the expression of NLRP3,ASC,caspase-1,and ICAM-1 was detected by Western blotting.RESULTS In vivo experiments confirmed that Pae could effectively reduce serum TC and TG levels and decrease serum IL-1βand IL-6 levels,which demonstrated that Pae restricted AS develop⁃ment in hyperlipidemia rats.Both CCK-8 and trypan blue staining showed that the survival rate of RAECs in the Pae-Exos co-incubation group was higher than that in the model group.We also confirmed via real-time qPCR that Pae-Exos suppressed the expression of the inflammatory cytokines IL-1βand IL-6.Accordingly,Pae-Exos dose-dependently increased the survival rate of RAECs and reduced inflammatory response.Furthermore,compared with the model group,Pae-Exos more successfully increased the expression of miR-223 and inhibited IL-1βand IL-6 expression,which implied that Pae-exo may inhibited the inflammatory response of RAECs by increasing the content of miR-223.Subse⁃quently,we found that Pae-Exos reduced the expressions of NLRP3,ASC,caspase-1 and ICAM-1,which indicated that Pae-Exos may reduced RAECs inflammation by suppressing NLRP3 signaling pathway via promoting miR-223 expression.CONCLUSION Pae can inhibit the downstream NLRP3 inflammatory corpuscle signaling pathway by increasing the level of miR-223 in plasma Exos of hyperlipidemic rats,providing new insights into the anti-atherosclerosis activity of Pae.
