Aim Salidroside （SAL） is a phenylpropanoid glycoside isolated from the medicinal plant Rhodiola rosea. A recent study has reported that SAL can efficiently decrease atherosclerotic plaque formation in low-density li...Aim Salidroside （SAL） is a phenylpropanoid glycoside isolated from the medicinal plant Rhodiola rosea. A recent study has reported that SAL can efficiently decrease atherosclerotic plaque formation in low-density lipoprotein receptor - deficient mice. This study was to investigate the molecular mechanism of antiatherogenic effects of SAL. Method Six-week old apoE-/- male mice were fed a high-fat diet for 8 weeks and then were ad- ministered with SAL for another 8 weeks. Atherosclerotic lesion and vascular function were analyzed. Primary cul- tured human umbilical vein endothelial cells （HUVECs） were prepared. Superoxide anion （O2^-）, NO produc- tion, mitochondrial membrane potential （△ψm） and intracellular ATP and AMP levels were measured. Expression of eNOS and AMPK were analyzed by Western blot. Result SAL significantly improved endothelial function asso- ciated with increasing eNOS activation thus reduced the atherosclerotic lesion area. SAL increased eNOS-Serl177 phosphorylation and decreased eNOS-Thr495 phosphorylation. SAL significantly activated AMP-activated protein ki- nase （AMPK）. Both AMPK inhibitor and AMPK small interfering RNA （siRNA） abolished SAL-induced Akt- Ser473 and eNOS-Serl177 phosphorylation. In contrast, LY294002, the PI3k/Akt pathway inhibitor, abolished SAL-induced phosphorylation and expression of eNOS. SAL decreased cellular ATP content and increased the cel- lular AMP/ATP ratio, which was associated with the activation of AMPK. SAL was found to decrease A^m, which is likely consequence of reduced ATP production. Conclusion The action of SAL to reduce atherosclerotic lesion formation may at least be attributed to its effect on improving endothelial function by promoting nitric oxide （NO） production, which was associated with mitochondria depolarization and subsequent activation of the AMPK/PI3 IC/ Akt/eNOS pathway. Taken together, our data described the effects of SAL on mitochondria, which played critical roles in improving endothelial function in atherosclerosis.展开更多
OBJECTIVE To investigate the beneficial effect of berberine(BBR)on atherosclerosisin Apo^(-/-) E mice and explore the underlying mechanisms based on attenuating vascular inflammation and modulating calcification in hu...OBJECTIVE To investigate the beneficial effect of berberine(BBR)on atherosclerosisin Apo^(-/-) E mice and explore the underlying mechanisms based on attenuating vascular inflammation and modulating calcification in human umbilical vein endothelial cells(HUVECs) and smooth muscle cells(SMCs).METHODS 48 Apo-/-E mice,at 6-8 weeks old,were randomly allocated into 4 groups:normal,model,bbr and atorvastatin(positive control) groups with 12 mice in each group.They were fed with high-fat diet for 4 weeks except those in Normal group and then treated with indicated drugs orsolvent for another 4 weeks.The morphology and inflammation infiltration of aortic were examined with HE staining.The expression of BMP-2 in aortic was examined by immumohistochemical staining.Blood lipid levels were examined by automatic biochemical analyzer.The expression of IL-6,TNF-α and BMP-2 in serum and tissues was detected by ELISA method.The expression of ALP and the content of calcium were detected by commercially-available kits.HUVEC cells were stimulated with TNF-α and incubated with various concentrations of BBR for 24 h.The contents of intercellular cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule(VCAM-1),matrix metalloprotein-9(MMP-9) in the culture supernatant were detected by ELISA method.Calcification was induced with β-glycerophosphatein SMC cells and the effect of BBR on the content of calcium was examined.RESULTS 4-week berberine treatment markedly lowered serum TC and LDL-c levels and improved the plaque stability in Apo-/-E mice fed with a high-fat diet(P<0.05 or P<0.01) which was comparable with the effect of atorvastatin.Berberineal so significantly decreased the levels of IL-6 and TNF-α in mice serum and aortic tissues(P<0.05 or P<0.001).Berberine tended to decrease ALP,BMP-2 levels and the content of calcium in mice serum and aortic tissues(P<0.05,P<0.01 or P<0.001) which were not observed in atorvastatin group.Berberine significantly lowered the levels of ICAM-1,VCAM-1,and MMP-9 in TNF-α-stimulated HUVECs.It can also lowered the content of calcium in SMCs.CONCLUSION BBR can profitably regulate the levels of blood lipid in mice fed with a high-fat diet,decrease the injury caused by inflammation,and attenuate vascular calcification.It may improve atherosclerosis and play a role in cardiovascular protection.展开更多
Aim The expression of α3 subunit of nicotinic acetylcholine receptor （α3-nAChR） has been demonstra- ted in aorta, adipocyte and macrophage. The objective of the present study was to verify the regulatory roles of ...Aim The expression of α3 subunit of nicotinic acetylcholine receptor （α3-nAChR） has been demonstra- ted in aorta, adipocyte and macrophage. The objective of the present study was to verify the regulatory roles of α3- nAChR in the inflammatory responses of atherosclerosis. Methods The inflammatory indicators were detected in mouse macrophage, adipocytes and mouse aortic endothelial cells （MAECs） after the α3-nAChR was antagonized or after the α3-nAChR gene was silenced. Meanwhile, atherogenesis was induced in the apolipoprotein E knock-out （ ApoE^ -/- ） mice after fed with an atherogenic high-fat diet for 7 weeks. Results In MAECs, the lipopolysaccha- ride （LPS）-stimulated secretions of the adhesion molecules and inflammatory cytokines were significantly enhanced （30%± 80% ） after pretreatment with α-Conotoxin MII （an antagonist for α3-nAChR） or after knock-down with α3-nAChR gene. In adipocytes, the knock-down of α3 gene promoted the generations of the proin？ ammatory adi- pokines or cytokines but decreased the production of adiponectin, an anti-inflammatory adipokine, by 29.29 ± 9.43%. In macrophage silenced with α3-nAChR gene, the M1 （classical） activation was predominantly stimula- ted, whereas the M2 （alternative） activation was suppressed. In addition, the amount of the atherosclerotic lesions and the infiltration of the M1 type activated macrophages into the arterial wall were markedly elevated in the α- Conotoxin MII-treated ApoE -/- mice. Conclusion The α3-nAChR may play a pivotal role in regulating the atherogenesis through influencing the inflammatory responses of ECs, macrophages and adipocytes. The mecha- nisms involve the regulations of multiple cell signaling pathways.展开更多
Atherosclerosis is the most common cause of cardiovascular diseases, such as myocardial infarction and stroke. The aim of this study was to investigate the effects of a novel compound ZBM30 on atherosclerosis in ApoE-...Atherosclerosis is the most common cause of cardiovascular diseases, such as myocardial infarction and stroke. The aim of this study was to investigate the effects of a novel compound ZBM30 on atherosclerosis in ApoE- deficient mice and its associated mechanism. ApoE-deficient mice （6 weeks old）, fed an atherogenic high-fat and high cholesterol diet for 8 weeks, were divided into three groups. Two groups were orally administrated ZBM30 （10, 30 nag ~ kg-1） daily for 12 weeks, while the control group was administered saline. Atherosclerotic lesions with en face aortas were evaluated by Sudan IV staining, and lesion areas in aortic sinuses were evaluated by oil red O staining. Necrotic core areas and fibrous cap areas in the lesion were evaluated by henaatoxylin and eosin （HE） staining and Masson＇ s trichronae staining in the aorta sinuses. The effects of ZBM30 on cholesterol accumulation in naacrophages and cholesterol transporters： ATP binding cassette A1 （ABCA1） and ATP binding cassette G1 （AB- CG1） were evaluated by oil red O assay, 3H-cholesterol efflux assay, Western blot, and real-time PCR on macro- phage cell lines： Raw 264.7 and THP-1. Inanauno-fluoresces was used to determine the ABCA1 expression in naac- rophage in aorta sinuses. Luciferase reporters of wild type and mutant types of ABCA1 promoter were constructed to determine the regulatory domain of ZBM30 on ABCA1 promoter. Results showed that, compared with the control group, en face lesions in ZBM30 group （ 10, 30 mg · kg^-1 ） were reduced 54.96 ± 10.06% and 71.50 ± 15.37% respectively, and aorta sinus lesions were reduced 41.85 ± 11.21% and 82.23 ± 8.25% respectively. Necrotic core areas in the ZBM30 group were markedly reduced and fibrous cap areas were not changed. Oil red O staining and 3 H-cholesterol efflux assays on Raw 264.7 cell line revealed that ZBM30 significantly attenuated the cholesterol accumulation in naacrophages by enhancing apolipoprotein AI and HDL mediated cholesterol efflux. Furthermore, ZBM30 induced the protein and naRNA expression of cholesterol transporters such as ABCA1 and ABCG1. Inanauno- fluoresces experiment revealed that ZBM30 induced the ABCA1 expression in naacrophage in the lesion, which is consistent with the results in vitro. Luciferase reporter assay revealed that ZBM30 exerted its effect on ABCA1 via liver X receptor （LXR） binding domain. In conclusion, ZBM30 suppresses atherosclerosis through up-regulating cholesterol efflux via ABCA1 and ABCG1 transporters in ApoE-deficient mice.展开更多
OBJECTIVE To investigate the anti-pyroptotic effects of melatonin in atherosclerotic endothelium and to elucidate the potential mechanisms.METHODS ApoE-/-mice were randomly divid.ed into four groups(n=8):the normal-di...OBJECTIVE To investigate the anti-pyroptotic effects of melatonin in atherosclerotic endothelium and to elucidate the potential mechanisms.METHODS ApoE-/-mice were randomly divid.ed into four groups(n=8):the normal-diet group(ND),the normal-diet group treated with melatonin(10 mg·kg^(-1))(ND+MLT),the high-fat-diet group(HFD),and the high-fat-diet group treated with melatonin(HFD+MLT).After 12 weeks,the expression levels of pyroptosis related genes including NLRP3,ASC,cleaved-caspase 1,GSDMD-N,IL-1β and IL-18 were examined in aortic endothelium by Western blotting,qRT-PCR and immunofluorescent staining.Besides,levels of MEG3 and miR-223 were also tested by qRT-PCR.The interaction between MEG3 and miR-223 was detected by luciferase assay.For in vitro study,human aortic endothelial cells(HAECs) were transiently transfected with miR-223 mimic,miR-223 inhibitor(AMO-223),MEG3-overexpressing plasmid or negative controls.After 6 h of transfection,the medium was replaced by fresh medium with or without ox-LDL(25 μg·mL-1) for 24 h and then treated with or without melatonin(10 μmol·L^(-1)) for 48 h.Cell pyroptosis was evaluated by Hoechst 33342/PI staining and differentially expressed pyroptosis related genes.RESULTS Melatonin markedly reduced the atherosclerotic plaque in aorta of ApoE-/-mice.Meanwhile,melatonin also attenuated the expression NLRP3,ASC,cleaved-caspase1,NF-κB/GSDMD,GSDMD-N termini,IL-1β,and IL-18 in aortic endo.thelium.Consistent anti-pyroptotic effects were also observed in ox-LDL-treated HAECs.We found that lncRNAMEG3 enhanced pyroptosis in HAECs.Moreover,MEG3 acted as an endogenous sponge by sequence complementarity to suppress the function of miR-223 and to increase NLRP3 expression and enhance endothelial cell pyroptosis.Furthermore,knockdown of miR-223 blocked the anti-pyroptotic actions of melatonin in ox-LDL-treated HAECs.CONCLUSION Melatonin prevents endothelial cell pyroptosis via MEG3/miR-223/NLRP3 axis in atherosclerosis and therefore melatonin replacement might be considered a new strategy for protecting endothelium against pyroptosis thereby for the treat.ment of atherosclerosis associated with pyroptosis.展开更多
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
Apelin, the endogenous ligand of APJ which is a member of G protein-coupled receptors, expressed in a variety of tissues in vivo and involved in the physiological and pathological processes. Studies had shown that ape...Apelin, the endogenous ligand of APJ which is a member of G protein-coupled receptors, expressed in a variety of tissues in vivo and involved in the physiological and pathological processes. Studies had shown that ape- lin/APJ system was involved in the regulation of cardiovascular function, insulin sensitivity and had a close connec- tion with oxidative stress and inflammatory cytokines. Therefore, this article summarized that apelin/APJ system such as atherosclerosis, hyperten- had the effects of inflammation-related diseases associated with oxidative stress, ischemia-reperfusion injury, tumour and pre-eclampsia, etc. sion, diabetes and its mierovaseular complications, Meanwhile the drugs targeted apelin/APJ was introduced in order to provide the novel therapeutic strategy for in- flammation-related diseases.展开更多
Background Cardiovascular disease has become a major cause of death worldwide.Atherosclerosis is the pathological basis of many cardiovascular diseases.It is well established that hemodynamics also play an important r...Background Cardiovascular disease has become a major cause of death worldwide.Atherosclerosis is the pathological basis of many cardiovascular diseases.It is well established that hemodynamics also play an important role in endothelial cell mediated atherosclerotic development.In the process of inflammatory reaction,the damage of vascular endothelial cells is the initial link,and various factors can cause damage of endothelial cells.The change of shear stress is considered to be one of the important factors.In the body,vascular endothelial cells are constantly exposed to blood flow.Flow conditions critically regulate endothelial cell functions in the vasculature.Shear stress not only influences the endothelial cell morphology,migration,differentiation and proliferation,but also regulates the expression of proteins in the endothelial cells.Reduced shear stress resulting from disturbed blood flow can drive the development of vascular inflammatory lesions and promote the formation of atherosclerosis.In the present study,the objective of our study is the comprehensive identification of CX3CR1/NF-κB signaling pathway involved in low shear stress(LSS)-induced inflammation in HUVECs through protein profiling and cell function experiment.Methods Human umbilical vein endothelial cell was cultured onglass slides and placed in a parallel plate flow chamber.M199 culture medium was used for low laminar shear stress at 4.14 dyn/cm2,2 h for the testing group,CX3CR1 sh-RNA and NF-κB inhibitor PDTC are used to block the effects of CX3CR1 and P65.The expression levels of the protein were determined by western blot analysis.Mononuclear cell adhesion assays and scratch assays are used to detect cell adhesion and migration.Results Western blotting analyses revealed that compared with the controls,there is a significant increase in the expression of CX3CR1,nucleusP65,intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1)and Interleukin-6(IL-6),while the expression of cytosolic P65 and IκB was significantly reduced in human umbilical vein endothelial cells(HUVECs)treated with LSS.CX3CR1 Sh-RNA was use to reveal its effect on LSS-induced inflammation.Further,specific NF-κB P65 inhibitors(PDTC)were used to reveal the downstream NF-κB P65 exclusively involved in LSS-induced inflammation in HUVECs,this effect can be abrogated by CX3CR1 sh-RNA and NF-κB inhibitors.Monocyte adhesion assay and scratch test revealed LSS promotes adhesion of monocytes and migration of cells,this effect can be abrogated by CX3CR1 sh-RNA and NF-κB inhibitors.LSS is involved in the expression of adhesion moleculesand chemokines,which are important for the initiation of endothelial inflammation-related atherosclerosis.Conclusions The activation of CX3CR1/NF-κB signaling pathway induced by low shear stress in endothelial cells may lead to the future therapeutic targets of atherosclerotic inflammation.展开更多
OBJECTIVE To reveal the under mechanism of how Liuwei Dihuang(LWDH)inhibits the phenotypic con⁃version of VSMCs.METHODS 24 ApoE-/-mice were divided into 4 groups:sham group,model group,E2 group,and LWDH group.Six C57B...OBJECTIVE To reveal the under mechanism of how Liuwei Dihuang(LWDH)inhibits the phenotypic con⁃version of VSMCs.METHODS 24 ApoE-/-mice were divided into 4 groups:sham group,model group,E2 group,and LWDH group.Six C57BN/L6 mice were used as control group.The primary VSMCs were divided into control group,model group,E2 group,LWDH group,LWDH+MPP group,and LWDH+PHTPP group with or without control siRNA,ERαsiRNA,ERβsiRNA,and myocardin siRNA.Oil red staining was used to evaluate the lipid deposition in the cardiac aorta.Serum chemistry analysis to test serum TG,TC,LDL,and HDL.Immunofluorescence staining was used to testα-SMA,osteopontin and F-actin.Immunohistochemical staining was performed to check out the myocardin in the cardiac aorta.The mRNA levels ofα-SMA,osteopontin,ERα,ERβ,SRC3 and myocardin were detected by real time-PCR,and the protein expression levels of them were detected by Western blotting.Co-immunoprecipitation was proceeded to test the interaction between ERαand SRC3 and SRC3 and myocardin.Flow cytometry was used to check out the cell cycle.Wound healing assay and Transwell were managed to evaluate the migration capacity of VSMCs.RESULTS In vivo administration of LWDH suppressed atherosclerotic symptoms,decreases phenotypic marker of vascular endothelial cell,and increases phenotypic marker of VSMC in ovariectomized ApoE-/-female mice.Moreover,LWDH significantly increased the mRNA and protein expression levels of ERα,ERβ,SRC3 and myocardin in the cardiac aorta of ovariecto⁃mized ApoE-/-female mice.In vitro,LWDH altered cell cycle and reduced the elevated cyclinD protein expression migra⁃tion capacity and in the model VSMCs.In addition,LWDH inhibited phenotypic conversion and promoted the expression of ER,SRC3,and myocardin of the primary VSMC phenotypic conversion model.Inhibition of ERαalmost completely eliminated the impacts of LWDH onα-SMA and osteopontin.Furthermore,LWDH promoted the interaction between ERαand SRC3 and up-regulated the co-activation of SRC3 and myocardin.CONCLUSION LWDH could inhibit the pheno⁃typic conversion of VSMCs in vitro and in vivo by increasing the activity of myocardin through up-regulating the expres⁃sion of ERαand promoting the interaction between ERαand SRC3.Our research reveals the under mechanism of how LWDH inhibits the phenotypic conversion of VSMCs.展开更多
Objective Atherosclerotic lesions preferentially occur at branch points of arterial trees where the blood flow is disturbed.Disturbed flow increases endothelial permeability,vascular barrier dysfunction,and finally th...Objective Atherosclerotic lesions preferentially occur at branch points of arterial trees where the blood flow is disturbed.Disturbed flow increases endothelial permeability,vascular barrier dysfunction,and finally the development of atherosclerosis.CTRP1,a member of C1q/TNF related protein(CTRP)family,is a novel secreted glycoprotein and its biological functions are largely undefined.展开更多
文摘Aim Salidroside （SAL） is a phenylpropanoid glycoside isolated from the medicinal plant Rhodiola rosea. A recent study has reported that SAL can efficiently decrease atherosclerotic plaque formation in low-density lipoprotein receptor - deficient mice. This study was to investigate the molecular mechanism of antiatherogenic effects of SAL. Method Six-week old apoE-/- male mice were fed a high-fat diet for 8 weeks and then were ad- ministered with SAL for another 8 weeks. Atherosclerotic lesion and vascular function were analyzed. Primary cul- tured human umbilical vein endothelial cells （HUVECs） were prepared. Superoxide anion （O2^-）, NO produc- tion, mitochondrial membrane potential （△ψm） and intracellular ATP and AMP levels were measured. Expression of eNOS and AMPK were analyzed by Western blot. Result SAL significantly improved endothelial function asso- ciated with increasing eNOS activation thus reduced the atherosclerotic lesion area. SAL increased eNOS-Serl177 phosphorylation and decreased eNOS-Thr495 phosphorylation. SAL significantly activated AMP-activated protein ki- nase （AMPK）. Both AMPK inhibitor and AMPK small interfering RNA （siRNA） abolished SAL-induced Akt- Ser473 and eNOS-Serl177 phosphorylation. In contrast, LY294002, the PI3k/Akt pathway inhibitor, abolished SAL-induced phosphorylation and expression of eNOS. SAL decreased cellular ATP content and increased the cel- lular AMP/ATP ratio, which was associated with the activation of AMPK. SAL was found to decrease A^m, which is likely consequence of reduced ATP production. Conclusion The action of SAL to reduce atherosclerotic lesion formation may at least be attributed to its effect on improving endothelial function by promoting nitric oxide （NO） production, which was associated with mitochondria depolarization and subsequent activation of the AMPK/PI3 IC/ Akt/eNOS pathway. Taken together, our data described the effects of SAL on mitochondria, which played critical roles in improving endothelial function in atherosclerosis.
基金supported by National Science Foundation of China(81402943)CAMS Major Collaborative Innovation Project(2016-I2M-1-011)PUMC Youth Fund(3332015168)
文摘OBJECTIVE To investigate the beneficial effect of berberine(BBR)on atherosclerosisin Apo^(-/-) E mice and explore the underlying mechanisms based on attenuating vascular inflammation and modulating calcification in human umbilical vein endothelial cells(HUVECs) and smooth muscle cells(SMCs).METHODS 48 Apo-/-E mice,at 6-8 weeks old,were randomly allocated into 4 groups:normal,model,bbr and atorvastatin(positive control) groups with 12 mice in each group.They were fed with high-fat diet for 4 weeks except those in Normal group and then treated with indicated drugs orsolvent for another 4 weeks.The morphology and inflammation infiltration of aortic were examined with HE staining.The expression of BMP-2 in aortic was examined by immumohistochemical staining.Blood lipid levels were examined by automatic biochemical analyzer.The expression of IL-6,TNF-α and BMP-2 in serum and tissues was detected by ELISA method.The expression of ALP and the content of calcium were detected by commercially-available kits.HUVEC cells were stimulated with TNF-α and incubated with various concentrations of BBR for 24 h.The contents of intercellular cell adhesion molecule-1(ICAM-1),vascular cell adhesion molecule(VCAM-1),matrix metalloprotein-9(MMP-9) in the culture supernatant were detected by ELISA method.Calcification was induced with β-glycerophosphatein SMC cells and the effect of BBR on the content of calcium was examined.RESULTS 4-week berberine treatment markedly lowered serum TC and LDL-c levels and improved the plaque stability in Apo-/-E mice fed with a high-fat diet(P<0.05 or P<0.01) which was comparable with the effect of atorvastatin.Berberineal so significantly decreased the levels of IL-6 and TNF-α in mice serum and aortic tissues(P<0.05 or P<0.001).Berberine tended to decrease ALP,BMP-2 levels and the content of calcium in mice serum and aortic tissues(P<0.05,P<0.01 or P<0.001) which were not observed in atorvastatin group.Berberine significantly lowered the levels of ICAM-1,VCAM-1,and MMP-9 in TNF-α-stimulated HUVECs.It can also lowered the content of calcium in SMCs.CONCLUSION BBR can profitably regulate the levels of blood lipid in mice fed with a high-fat diet,decrease the injury caused by inflammation,and attenuate vascular calcification.It may improve atherosclerosis and play a role in cardiovascular protection.
文摘Aim The expression of α3 subunit of nicotinic acetylcholine receptor （α3-nAChR） has been demonstra- ted in aorta, adipocyte and macrophage. The objective of the present study was to verify the regulatory roles of α3- nAChR in the inflammatory responses of atherosclerosis. Methods The inflammatory indicators were detected in mouse macrophage, adipocytes and mouse aortic endothelial cells （MAECs） after the α3-nAChR was antagonized or after the α3-nAChR gene was silenced. Meanwhile, atherogenesis was induced in the apolipoprotein E knock-out （ ApoE^ -/- ） mice after fed with an atherogenic high-fat diet for 7 weeks. Results In MAECs, the lipopolysaccha- ride （LPS）-stimulated secretions of the adhesion molecules and inflammatory cytokines were significantly enhanced （30%± 80% ） after pretreatment with α-Conotoxin MII （an antagonist for α3-nAChR） or after knock-down with α3-nAChR gene. In adipocytes, the knock-down of α3 gene promoted the generations of the proin？ ammatory adi- pokines or cytokines but decreased the production of adiponectin, an anti-inflammatory adipokine, by 29.29 ± 9.43%. In macrophage silenced with α3-nAChR gene, the M1 （classical） activation was predominantly stimula- ted, whereas the M2 （alternative） activation was suppressed. In addition, the amount of the atherosclerotic lesions and the infiltration of the M1 type activated macrophages into the arterial wall were markedly elevated in the α- Conotoxin MII-treated ApoE -/- mice. Conclusion The α3-nAChR may play a pivotal role in regulating the atherogenesis through influencing the inflammatory responses of ECs, macrophages and adipocytes. The mecha- nisms involve the regulations of multiple cell signaling pathways.
文摘Atherosclerosis is the most common cause of cardiovascular diseases, such as myocardial infarction and stroke. The aim of this study was to investigate the effects of a novel compound ZBM30 on atherosclerosis in ApoE- deficient mice and its associated mechanism. ApoE-deficient mice （6 weeks old）, fed an atherogenic high-fat and high cholesterol diet for 8 weeks, were divided into three groups. Two groups were orally administrated ZBM30 （10, 30 nag ~ kg-1） daily for 12 weeks, while the control group was administered saline. Atherosclerotic lesions with en face aortas were evaluated by Sudan IV staining, and lesion areas in aortic sinuses were evaluated by oil red O staining. Necrotic core areas and fibrous cap areas in the lesion were evaluated by henaatoxylin and eosin （HE） staining and Masson＇ s trichronae staining in the aorta sinuses. The effects of ZBM30 on cholesterol accumulation in naacrophages and cholesterol transporters： ATP binding cassette A1 （ABCA1） and ATP binding cassette G1 （AB- CG1） were evaluated by oil red O assay, 3H-cholesterol efflux assay, Western blot, and real-time PCR on macro- phage cell lines： Raw 264.7 and THP-1. Inanauno-fluoresces was used to determine the ABCA1 expression in naac- rophage in aorta sinuses. Luciferase reporters of wild type and mutant types of ABCA1 promoter were constructed to determine the regulatory domain of ZBM30 on ABCA1 promoter. Results showed that, compared with the control group, en face lesions in ZBM30 group （ 10, 30 mg · kg^-1 ） were reduced 54.96 ± 10.06% and 71.50 ± 15.37% respectively, and aorta sinus lesions were reduced 41.85 ± 11.21% and 82.23 ± 8.25% respectively. Necrotic core areas in the ZBM30 group were markedly reduced and fibrous cap areas were not changed. Oil red O staining and 3 H-cholesterol efflux assays on Raw 264.7 cell line revealed that ZBM30 significantly attenuated the cholesterol accumulation in naacrophages by enhancing apolipoprotein AI and HDL mediated cholesterol efflux. Furthermore, ZBM30 induced the protein and naRNA expression of cholesterol transporters such as ABCA1 and ABCG1. Inanauno- fluoresces experiment revealed that ZBM30 induced the ABCA1 expression in naacrophage in the lesion, which is consistent with the results in vitro. Luciferase reporter assay revealed that ZBM30 exerted its effect on ABCA1 via liver X receptor （LXR） binding domain. In conclusion, ZBM30 suppresses atherosclerosis through up-regulating cholesterol efflux via ABCA1 and ABCG1 transporters in ApoE-deficient mice.
基金supported by National Natural Science Foundation of China(8157039981270042)
文摘OBJECTIVE To investigate the anti-pyroptotic effects of melatonin in atherosclerotic endothelium and to elucidate the potential mechanisms.METHODS ApoE-/-mice were randomly divid.ed into four groups(n=8):the normal-diet group(ND),the normal-diet group treated with melatonin(10 mg·kg^(-1))(ND+MLT),the high-fat-diet group(HFD),and the high-fat-diet group treated with melatonin(HFD+MLT).After 12 weeks,the expression levels of pyroptosis related genes including NLRP3,ASC,cleaved-caspase 1,GSDMD-N,IL-1β and IL-18 were examined in aortic endothelium by Western blotting,qRT-PCR and immunofluorescent staining.Besides,levels of MEG3 and miR-223 were also tested by qRT-PCR.The interaction between MEG3 and miR-223 was detected by luciferase assay.For in vitro study,human aortic endothelial cells(HAECs) were transiently transfected with miR-223 mimic,miR-223 inhibitor(AMO-223),MEG3-overexpressing plasmid or negative controls.After 6 h of transfection,the medium was replaced by fresh medium with or without ox-LDL(25 μg·mL-1) for 24 h and then treated with or without melatonin(10 μmol·L^(-1)) for 48 h.Cell pyroptosis was evaluated by Hoechst 33342/PI staining and differentially expressed pyroptosis related genes.RESULTS Melatonin markedly reduced the atherosclerotic plaque in aorta of ApoE-/-mice.Meanwhile,melatonin also attenuated the expression NLRP3,ASC,cleaved-caspase1,NF-κB/GSDMD,GSDMD-N termini,IL-1β,and IL-18 in aortic endo.thelium.Consistent anti-pyroptotic effects were also observed in ox-LDL-treated HAECs.We found that lncRNAMEG3 enhanced pyroptosis in HAECs.Moreover,MEG3 acted as an endogenous sponge by sequence complementarity to suppress the function of miR-223 and to increase NLRP3 expression and enhance endothelial cell pyroptosis.Furthermore,knockdown of miR-223 blocked the anti-pyroptotic actions of melatonin in ox-LDL-treated HAECs.CONCLUSION Melatonin prevents endothelial cell pyroptosis via MEG3/miR-223/NLRP3 axis in atherosclerosis and therefore melatonin replacement might be considered a new strategy for protecting endothelium against pyroptosis thereby for the treat.ment of atherosclerosis associated with pyroptosis.
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.
文摘Apelin, the endogenous ligand of APJ which is a member of G protein-coupled receptors, expressed in a variety of tissues in vivo and involved in the physiological and pathological processes. Studies had shown that ape- lin/APJ system was involved in the regulation of cardiovascular function, insulin sensitivity and had a close connec- tion with oxidative stress and inflammatory cytokines. Therefore, this article summarized that apelin/APJ system such as atherosclerosis, hyperten- had the effects of inflammation-related diseases associated with oxidative stress, ischemia-reperfusion injury, tumour and pre-eclampsia, etc. sion, diabetes and its mierovaseular complications, Meanwhile the drugs targeted apelin/APJ was introduced in order to provide the novel therapeutic strategy for in- flammation-related diseases.
基金supported by a project grant from the National Natural Science Foundation of China ( 31860261)
文摘Background Cardiovascular disease has become a major cause of death worldwide.Atherosclerosis is the pathological basis of many cardiovascular diseases.It is well established that hemodynamics also play an important role in endothelial cell mediated atherosclerotic development.In the process of inflammatory reaction,the damage of vascular endothelial cells is the initial link,and various factors can cause damage of endothelial cells.The change of shear stress is considered to be one of the important factors.In the body,vascular endothelial cells are constantly exposed to blood flow.Flow conditions critically regulate endothelial cell functions in the vasculature.Shear stress not only influences the endothelial cell morphology,migration,differentiation and proliferation,but also regulates the expression of proteins in the endothelial cells.Reduced shear stress resulting from disturbed blood flow can drive the development of vascular inflammatory lesions and promote the formation of atherosclerosis.In the present study,the objective of our study is the comprehensive identification of CX3CR1/NF-κB signaling pathway involved in low shear stress(LSS)-induced inflammation in HUVECs through protein profiling and cell function experiment.Methods Human umbilical vein endothelial cell was cultured onglass slides and placed in a parallel plate flow chamber.M199 culture medium was used for low laminar shear stress at 4.14 dyn/cm2,2 h for the testing group,CX3CR1 sh-RNA and NF-κB inhibitor PDTC are used to block the effects of CX3CR1 and P65.The expression levels of the protein were determined by western blot analysis.Mononuclear cell adhesion assays and scratch assays are used to detect cell adhesion and migration.Results Western blotting analyses revealed that compared with the controls,there is a significant increase in the expression of CX3CR1,nucleusP65,intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1)and Interleukin-6(IL-6),while the expression of cytosolic P65 and IκB was significantly reduced in human umbilical vein endothelial cells(HUVECs)treated with LSS.CX3CR1 Sh-RNA was use to reveal its effect on LSS-induced inflammation.Further,specific NF-κB P65 inhibitors(PDTC)were used to reveal the downstream NF-κB P65 exclusively involved in LSS-induced inflammation in HUVECs,this effect can be abrogated by CX3CR1 sh-RNA and NF-κB inhibitors.Monocyte adhesion assay and scratch test revealed LSS promotes adhesion of monocytes and migration of cells,this effect can be abrogated by CX3CR1 sh-RNA and NF-κB inhibitors.LSS is involved in the expression of adhesion moleculesand chemokines,which are important for the initiation of endothelial inflammation-related atherosclerosis.Conclusions The activation of CX3CR1/NF-κB signaling pathway induced by low shear stress in endothelial cells may lead to the future therapeutic targets of atherosclerotic inflammation.
基金National Natural Science Foundation of China(8177319081774029)Jiangsu Provincial Science and Technology Department Social Development Fund(BE2011846)
文摘OBJECTIVE To reveal the under mechanism of how Liuwei Dihuang(LWDH)inhibits the phenotypic con⁃version of VSMCs.METHODS 24 ApoE-/-mice were divided into 4 groups:sham group,model group,E2 group,and LWDH group.Six C57BN/L6 mice were used as control group.The primary VSMCs were divided into control group,model group,E2 group,LWDH group,LWDH+MPP group,and LWDH+PHTPP group with or without control siRNA,ERαsiRNA,ERβsiRNA,and myocardin siRNA.Oil red staining was used to evaluate the lipid deposition in the cardiac aorta.Serum chemistry analysis to test serum TG,TC,LDL,and HDL.Immunofluorescence staining was used to testα-SMA,osteopontin and F-actin.Immunohistochemical staining was performed to check out the myocardin in the cardiac aorta.The mRNA levels ofα-SMA,osteopontin,ERα,ERβ,SRC3 and myocardin were detected by real time-PCR,and the protein expression levels of them were detected by Western blotting.Co-immunoprecipitation was proceeded to test the interaction between ERαand SRC3 and SRC3 and myocardin.Flow cytometry was used to check out the cell cycle.Wound healing assay and Transwell were managed to evaluate the migration capacity of VSMCs.RESULTS In vivo administration of LWDH suppressed atherosclerotic symptoms,decreases phenotypic marker of vascular endothelial cell,and increases phenotypic marker of VSMC in ovariectomized ApoE-/-female mice.Moreover,LWDH significantly increased the mRNA and protein expression levels of ERα,ERβ,SRC3 and myocardin in the cardiac aorta of ovariecto⁃mized ApoE-/-female mice.In vitro,LWDH altered cell cycle and reduced the elevated cyclinD protein expression migra⁃tion capacity and in the model VSMCs.In addition,LWDH inhibited phenotypic conversion and promoted the expression of ER,SRC3,and myocardin of the primary VSMC phenotypic conversion model.Inhibition of ERαalmost completely eliminated the impacts of LWDH onα-SMA and osteopontin.Furthermore,LWDH promoted the interaction between ERαand SRC3 and up-regulated the co-activation of SRC3 and myocardin.CONCLUSION LWDH could inhibit the pheno⁃typic conversion of VSMCs in vitro and in vivo by increasing the activity of myocardin through up-regulating the expres⁃sion of ERαand promoting the interaction between ERαand SRC3.Our research reveals the under mechanism of how LWDH inhibits the phenotypic conversion of VSMCs.
文摘Objective Atherosclerotic lesions preferentially occur at branch points of arterial trees where the blood flow is disturbed.Disturbed flow increases endothelial permeability,vascular barrier dysfunction,and finally the development of atherosclerosis.CTRP1,a member of C1q/TNF related protein(CTRP)family,is a novel secreted glycoprotein and its biological functions are largely undefined.
