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 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
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 ~1H-NMR-based metabolomics approach was conducted to holistically explore the effect and mechanisms of Cydonia oblanga Mill flavonoids(COMF) on high-fat diet induced Atherosclerosis(AS) apoE-/-mice.METHODS A...OBJECTIVE ~1H-NMR-based metabolomics approach was conducted to holistically explore the effect and mechanisms of Cydonia oblanga Mill flavonoids(COMF) on high-fat diet induced Atherosclerosis(AS) apoE-/-mice.METHODS AS model was established on the apolipoprotein e knockout mice by high-fat diet.The ApoE-/-mice were split into 6 groups including control group,AS model group,COMF High dose(COMF-H) group,COMF medium dose(COMF-M) group,COMF Low dose(COMF-L) group and Simvastatin group as the positive control group.Serum samples from all groups were analyzed by ~1H-NMR technology and the OPLS-DA was conducted to distinguish the metabolic phenotypes.RESULTS Compared to the control group,serum levels of cholesterol,VLDL,leucine,isoleucine,valine,blood lipid,citrulline,methylamine,glucose,glycine,glycerol,myo-inositol,fructose,phenylalanine,unsaturated lipid,urea and other metabolites content significantly increased,while HDL,lactate,alanine,glutamate,glutamine,pyruvate,carnitine,citrate,choline content signifi.cantly decreased and the difference was statistically significant(P<0.05).The trend of metabolites in serum samples of COMF low,medium and high group was opposite to that of atherosclerosis model group and the difference was statistically significant(P<0.05).CONCLUSION Through functional analysis of these biomakers,amino acid metabolism,lipid metabolism,cholesterol metabolism,energy metabolism and inflammation reaction were considered as the most relevant pathological biomakers in the serum of AS mice.This study also demonstrates that COMF had the therapeutic effectiveness on AS through partly reversing the lipid,cholesterol,amino acid,energy metabolism and Inflammation reaction.展开更多
OBJECTIVE To explore the biomarkers and molecular mechanism of Huanglianjiedu decoction(HJD) on high fat diet-induced experimental atherosclerosis in rats.METHODS SD male rats were randomly dividedinto five groups(n=8...OBJECTIVE To explore the biomarkers and molecular mechanism of Huanglianjiedu decoction(HJD) on high fat diet-induced experimental atherosclerosis in rats.METHODS SD male rats were randomly dividedinto five groups(n=8):normal control group,model group,and three dosage groups(1.5,3 and 6 g crude drug per kilogram of body weight).Atherosclerosis was induced by the combination of regular intraperitoneal injection of vitamin D3 and high fat diet for 8 weeks.HJD was administered by oral gavage from the third week once per day and until the end of the study.After the final administration,the blood samples were collected for biochemical analyses [total cholesterol(TC),triglycerides(TG),highdensity lipoprotein(HDL-C),low-density cholesterol(LDL-C)] and blood gas analyses(PaO_2,PaCO_2,pH,ctHb,etc);the abdominal aorta sections were stained with hematoxylin and eosin for histopathology;the liver homogenate were determined for MDA,SOD,OX-LDL,MCP-1 and VCAM-1.The plasma samples were detected using ultraper formance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS).The data of endogenous compounds were preliminarily preprocessed by software Progenesis QI and then analyzed by multivari.ate statistical analysis software EZinfo 2.0 to screen the distinguished biomarkers and the metabolic pathways were analyzed through website http://www.metaboanalyst.ca/.RESULTS Compared with the normal control group,the content of TC,TG,LDL-C,PaCO_2,MDA,Ox-LDL,MCP-1 and VCAM-1 were significantly increased and HDL-C,PaO_2,ctHb and SOD decreased in the atherosclerosis rats.HJD could significantly attenuated the high fat-induced atherosclerosis pathological injury and the abovementioned indexes(P<0.05).The five groups could be clearly distinguished using the metabolomics method.The administration groups profile exhibited an apparent returning trend from that of the model group and that of the normal control group.Twenty-one endogenous metabolites has been significantly changed in atherosclerosis rats.HJD could remarkably up-regulate 5-L-glutamyl-taurine,L-beta-aspartylL-glutamic acid,histidinyl-hydroxyproline,tryptophyl-alanine,4′-O-methyl-(-)-epicatechin,and downregulate protoporphyrin IX,azelaic acid,lacto-N-triaose,cinnamoylglycine and 9′-carboxy-alpha-tocotri.enol.CONCLUSION The beneficial effect of HJD in high fat-induced atherosclerosis rats may be due to anti-oxidant and anti-inflammatory.And it is suggested that HJD may affect the model rats through tryptophan metabolism,taurine and hypotaurine metabolism,histidine metabolism,lysine degradation and porphyrin and chlorophyll metabolism pathway.展开更多
目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因...目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因敲除(apolipoprotein E knockout,APOE^(-/-))小鼠动脉粥样硬化(atherosclerosis,AS)。方法10只C57BL/6J小鼠作为空白对照组,60只APOE^(-/-)小鼠随机分为模型组、血管软化丸低剂量组、血管软化丸高剂量组、阿托伐他汀组、血管软化丸高剂量和AMPK抑制剂联合组、AMPK抑制剂组,给予高脂饮食饲养18周建造动脉粥样硬化小鼠模型,并于第13周给予不同方法干预,干预6周后取材,分别检测各组小鼠甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白-胆固醇(low density lipoprotein-eholesterol,LDL-C)、高密度脂蛋白-胆固醇(high density lipoprotein-eholesterol,HDL-C)水平,HE和油红O染色观察小鼠主动脉组织病理形态学变化,Elisa测定主动脉组织白细胞介素(interleukin,IL)-1β和IL-18水平,Western Blot检测AMPK、P-AMPK、NLRP3、裂解的半胱氨酸天冬酶1(Cleaved-Caspase-1)、Gasdermin D(GSDMD)-N蛋白表达,免疫荧光检测P-AMPK、NLRP3蛋白在主动脉根部的分布情况,透射电子显微镜观察主动脉内皮超微结构改变情况。结果与对照组相比,模型组大鼠血清TG、TC、LDL-C水平显著升高,HDL-C水平显著降低(P<0.01),主动脉根部细胞排列紊乱,且脂肪空泡明显,主动脉有明显粥样斑块沉积,P-AMPK蛋白表达水平显著降低,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平及IL-1β和IL-18水平显著升高(P<0.05或P<0.01);与模型组比较,血管软化丸高低剂量组及阿托伐他汀组血脂紊乱得到改善,主动脉组织及细胞病变程度减轻,P-AMPK蛋白表达水平显著升高,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平显著降低(P<0.05或P<0.01),IL-1β和IL-18水平显著降低(P<0.05或P<0.01);与血管软化丸高剂量+AMPK抑制剂组比较,血管软化丸高剂量组焦亡相关蛋白水平显著降低(P<0.05或P<0.01),主动脉内皮细胞损伤程度减轻,而AMPK抑制剂组与之相反。结论血管软化丸可能通过调控AMPK/NLRP3通路减轻细胞焦亡发挥对动脉粥样硬化小鼠的保护作用。展开更多
目的分析老年穿支动脉粥样硬化病患者血清微小RNA(micorRNA,miRNA)预测早期神经功能恶化的回归分析。方法选择2020年2月至2023年2月湖北医药学院附属随州市中心医院神经内科收治的老年穿支动脉粥样硬化病患者134例,依据早期神经功能恶...目的分析老年穿支动脉粥样硬化病患者血清微小RNA(micorRNA,miRNA)预测早期神经功能恶化的回归分析。方法选择2020年2月至2023年2月湖北医药学院附属随州市中心医院神经内科收治的老年穿支动脉粥样硬化病患者134例,依据早期神经功能恶化情况分为恶化组28例和未恶化组106例。入院时测定患者血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平,入院时及入院后7 d采用美国国立卫生研究院卒中量表(National Institutes of Health Stroke Scale,NIHSS)评分评估早期神经功能恶化情况。采用二元logistic回归分析法构建miR-130a、miR-210、miR-141-3p、miR-29a-3p预测老年穿支动脉粥样硬化病患者早期神经功能恶化模型,ROC曲线分析血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平对老年穿支动脉粥样硬化病患者早期神经功能恶化的预测价值。结果恶化组血清miR-130a、miR-210水平明显高于未恶化组,miR-141-3p、miR-29a-3p水平明显低于未恶化组,差异有统计学意义(P<0.01)。Logistic回归分析显示,血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平为老年穿支动脉粥样硬化病患者早期神经功能恶化的独立预测指标(P<0.05,P<0.01)。ROC曲线分析显示,血清miR-130a、miR-210、miR-141-3p、miR-29a-3p联合预测老年穿支动脉粥样硬化病患者早期神经功能恶化的曲线下面积为0.977(95%CI:0.936~0.995),敏感性为96.43%,特异性为90.57%,联合预测的效能明显优于各指标单独预测(P<0.01)。结论老年穿支动脉粥样硬化病患者血清miR-130a、miR-210、miR-141-3p、miR-29a-3p对预测早期神经功能恶化具有一定的价值,且四者联合检测可提高其预测效能。展开更多
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 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.
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(81660696) Research and development of new drugs Foundation of Xinjiang administration of traditional Chinese medicine(2017-02-05)
文摘OBJECTIVE ~1H-NMR-based metabolomics approach was conducted to holistically explore the effect and mechanisms of Cydonia oblanga Mill flavonoids(COMF) on high-fat diet induced Atherosclerosis(AS) apoE-/-mice.METHODS AS model was established on the apolipoprotein e knockout mice by high-fat diet.The ApoE-/-mice were split into 6 groups including control group,AS model group,COMF High dose(COMF-H) group,COMF medium dose(COMF-M) group,COMF Low dose(COMF-L) group and Simvastatin group as the positive control group.Serum samples from all groups were analyzed by ~1H-NMR technology and the OPLS-DA was conducted to distinguish the metabolic phenotypes.RESULTS Compared to the control group,serum levels of cholesterol,VLDL,leucine,isoleucine,valine,blood lipid,citrulline,methylamine,glucose,glycine,glycerol,myo-inositol,fructose,phenylalanine,unsaturated lipid,urea and other metabolites content significantly increased,while HDL,lactate,alanine,glutamate,glutamine,pyruvate,carnitine,citrate,choline content signifi.cantly decreased and the difference was statistically significant(P<0.05).The trend of metabolites in serum samples of COMF low,medium and high group was opposite to that of atherosclerosis model group and the difference was statistically significant(P<0.05).CONCLUSION Through functional analysis of these biomakers,amino acid metabolism,lipid metabolism,cholesterol metabolism,energy metabolism and inflammation reaction were considered as the most relevant pathological biomakers in the serum of AS mice.This study also demonstrates that COMF had the therapeutic effectiveness on AS through partly reversing the lipid,cholesterol,amino acid,energy metabolism and Inflammation reaction.
基金supported by National Natural Science Foundation of China(8170382381560744) Science and Technology Research Project of Jiangxi Provincial Education Department(GJJ170753)
文摘OBJECTIVE To explore the biomarkers and molecular mechanism of Huanglianjiedu decoction(HJD) on high fat diet-induced experimental atherosclerosis in rats.METHODS SD male rats were randomly dividedinto five groups(n=8):normal control group,model group,and three dosage groups(1.5,3 and 6 g crude drug per kilogram of body weight).Atherosclerosis was induced by the combination of regular intraperitoneal injection of vitamin D3 and high fat diet for 8 weeks.HJD was administered by oral gavage from the third week once per day and until the end of the study.After the final administration,the blood samples were collected for biochemical analyses [total cholesterol(TC),triglycerides(TG),highdensity lipoprotein(HDL-C),low-density cholesterol(LDL-C)] and blood gas analyses(PaO_2,PaCO_2,pH,ctHb,etc);the abdominal aorta sections were stained with hematoxylin and eosin for histopathology;the liver homogenate were determined for MDA,SOD,OX-LDL,MCP-1 and VCAM-1.The plasma samples were detected using ultraper formance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS).The data of endogenous compounds were preliminarily preprocessed by software Progenesis QI and then analyzed by multivari.ate statistical analysis software EZinfo 2.0 to screen the distinguished biomarkers and the metabolic pathways were analyzed through website http://www.metaboanalyst.ca/.RESULTS Compared with the normal control group,the content of TC,TG,LDL-C,PaCO_2,MDA,Ox-LDL,MCP-1 and VCAM-1 were significantly increased and HDL-C,PaO_2,ctHb and SOD decreased in the atherosclerosis rats.HJD could significantly attenuated the high fat-induced atherosclerosis pathological injury and the abovementioned indexes(P<0.05).The five groups could be clearly distinguished using the metabolomics method.The administration groups profile exhibited an apparent returning trend from that of the model group and that of the normal control group.Twenty-one endogenous metabolites has been significantly changed in atherosclerosis rats.HJD could remarkably up-regulate 5-L-glutamyl-taurine,L-beta-aspartylL-glutamic acid,histidinyl-hydroxyproline,tryptophyl-alanine,4′-O-methyl-(-)-epicatechin,and downregulate protoporphyrin IX,azelaic acid,lacto-N-triaose,cinnamoylglycine and 9′-carboxy-alpha-tocotri.enol.CONCLUSION The beneficial effect of HJD in high fat-induced atherosclerosis rats may be due to anti-oxidant and anti-inflammatory.And it is suggested that HJD may affect the model rats through tryptophan metabolism,taurine and hypotaurine metabolism,histidine metabolism,lysine degradation and porphyrin and chlorophyll metabolism pathway.
文摘目的探讨血管软化丸能否通过调控腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)通路抑制细胞焦亡改善载脂蛋白E基因敲除(apolipoprotein E knockout,APOE^(-/-))小鼠动脉粥样硬化(atherosclerosis,AS)。方法10只C57BL/6J小鼠作为空白对照组,60只APOE^(-/-)小鼠随机分为模型组、血管软化丸低剂量组、血管软化丸高剂量组、阿托伐他汀组、血管软化丸高剂量和AMPK抑制剂联合组、AMPK抑制剂组,给予高脂饮食饲养18周建造动脉粥样硬化小鼠模型,并于第13周给予不同方法干预,干预6周后取材,分别检测各组小鼠甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、低密度脂蛋白-胆固醇(low density lipoprotein-eholesterol,LDL-C)、高密度脂蛋白-胆固醇(high density lipoprotein-eholesterol,HDL-C)水平,HE和油红O染色观察小鼠主动脉组织病理形态学变化,Elisa测定主动脉组织白细胞介素(interleukin,IL)-1β和IL-18水平,Western Blot检测AMPK、P-AMPK、NLRP3、裂解的半胱氨酸天冬酶1(Cleaved-Caspase-1)、Gasdermin D(GSDMD)-N蛋白表达,免疫荧光检测P-AMPK、NLRP3蛋白在主动脉根部的分布情况,透射电子显微镜观察主动脉内皮超微结构改变情况。结果与对照组相比,模型组大鼠血清TG、TC、LDL-C水平显著升高,HDL-C水平显著降低(P<0.01),主动脉根部细胞排列紊乱,且脂肪空泡明显,主动脉有明显粥样斑块沉积,P-AMPK蛋白表达水平显著降低,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平及IL-1β和IL-18水平显著升高(P<0.05或P<0.01);与模型组比较,血管软化丸高低剂量组及阿托伐他汀组血脂紊乱得到改善,主动脉组织及细胞病变程度减轻,P-AMPK蛋白表达水平显著升高,NLRP3、Cleaved-Caspase 1、GSDMD-N蛋白表达水平显著降低(P<0.05或P<0.01),IL-1β和IL-18水平显著降低(P<0.05或P<0.01);与血管软化丸高剂量+AMPK抑制剂组比较,血管软化丸高剂量组焦亡相关蛋白水平显著降低(P<0.05或P<0.01),主动脉内皮细胞损伤程度减轻,而AMPK抑制剂组与之相反。结论血管软化丸可能通过调控AMPK/NLRP3通路减轻细胞焦亡发挥对动脉粥样硬化小鼠的保护作用。
文摘目的分析老年穿支动脉粥样硬化病患者血清微小RNA(micorRNA,miRNA)预测早期神经功能恶化的回归分析。方法选择2020年2月至2023年2月湖北医药学院附属随州市中心医院神经内科收治的老年穿支动脉粥样硬化病患者134例,依据早期神经功能恶化情况分为恶化组28例和未恶化组106例。入院时测定患者血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平,入院时及入院后7 d采用美国国立卫生研究院卒中量表(National Institutes of Health Stroke Scale,NIHSS)评分评估早期神经功能恶化情况。采用二元logistic回归分析法构建miR-130a、miR-210、miR-141-3p、miR-29a-3p预测老年穿支动脉粥样硬化病患者早期神经功能恶化模型,ROC曲线分析血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平对老年穿支动脉粥样硬化病患者早期神经功能恶化的预测价值。结果恶化组血清miR-130a、miR-210水平明显高于未恶化组,miR-141-3p、miR-29a-3p水平明显低于未恶化组,差异有统计学意义(P<0.01)。Logistic回归分析显示,血清miR-130a、miR-210、miR-141-3p、miR-29a-3p水平为老年穿支动脉粥样硬化病患者早期神经功能恶化的独立预测指标(P<0.05,P<0.01)。ROC曲线分析显示,血清miR-130a、miR-210、miR-141-3p、miR-29a-3p联合预测老年穿支动脉粥样硬化病患者早期神经功能恶化的曲线下面积为0.977(95%CI:0.936~0.995),敏感性为96.43%,特异性为90.57%,联合预测的效能明显优于各指标单独预测(P<0.01)。结论老年穿支动脉粥样硬化病患者血清miR-130a、miR-210、miR-141-3p、miR-29a-3p对预测早期神经功能恶化具有一定的价值,且四者联合检测可提高其预测效能。
