目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与...目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡程度的相关性。方法:将健康雄性新西兰兔34只,随机分成空白对照组(K组)和造模组(B组)。空白对照组8只,给予普通饲料喂养18周。造模组(B组)26只,给予高脂饲料喂养,实验开始第1周一次性注射牛血清白蛋白,实验第10周,随机抽取2只兔做病理检测证实模型组已形成粥样斑块。其余B组兔随机分为模型对照组、辛伐他汀组、健脑软脉方组。辛伐他汀组、健脑软脉方组分别予以辛伐他汀、健脑软脉方水煎剂干预,模型对照组予以生理盐水灌胃,持续8周,实验结束,于处死动物前24 h和48 h 2次给予药物触发易损斑块(中国斑点蝰蛇毒和组胺)。空白对照组(K组)按上述给药时间和部位给予同等剂量无菌生理盐水注射。实验终点处死实验兔,开胸迅速分离主动脉全长,检测各项指标。结果:模型对照组的主动脉病变程度、血脂水平、血清RANTES水平、斑块内RANTES m RAN和蛋白表达水平、VSMCs凋亡率与健脑软脉方组、辛伐他汀组、空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组与空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组之间无显著差异(P>0.05)。结论:本研究表明补肾活血通络法可能通过调节血脂水平,抑制炎症反应,抑制平滑肌细胞凋亡,从而抑制模型兔动脉粥样硬化斑块的不稳定性进展。展开更多
A new cerebroside,1-O-(β-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2′R)-2’-hydroxylignoceranoylamino]-4,8-tetradecene-3- diol was isolated from the 60%EtOH extract of traditional Chinese medical plant Cyperus rotundus...A new cerebroside,1-O-(β-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2′R)-2’-hydroxylignoceranoylamino]-4,8-tetradecene-3- diol was isolated from the 60%EtOH extract of traditional Chinese medical plant Cyperus rotundus L.Its structure was determined on the basis of spectroscopic data.This new compound showed anti-proliferation effect on vascular smooth muscle cells(VSMCs).展开更多
Background To study the molecular mechanisms of CREG(the cellular repressor of E1A-stimulated gene) on proliferation of VSMCs in vitro.Methods The pRc/CMV-CREG plasmid or the pSM2-siCREG plasmid was transferred into h...Background To study the molecular mechanisms of CREG(the cellular repressor of E1A-stimulated gene) on proliferation of VSMCs in vitro.Methods The pRc/CMV-CREG plasmid or the pSM2-siCREG plasmid was transferred into human vascular smooth muscle cells(hVSMCs) to produce the cell clone that over-expression or down-expression of CREG respectively.BrdU assay and FACS cell cycle analysis were used to detect the proliferation of cells.Western blotting and immunocytochemistry show the expression and localization of IGF2R in hVSMCs.RT-PCR and ELISA assay determined the expression and secretion of IGFII factor. Alex488-labeled rhIGFII was used to investigate the endocysis of cells.And the blockade of IGFII internalization by treatment both the neutralized antibody of anti-IGF2R and rsIGF2R detected the effect of IGFII on VSMCs growth. Furthermore,Western blotting and signal pathway inhibitor were used to analysis the activation of PI3K/AKT and ERK on VSMCs proliferation.Results Western blotting identified that the expression of CREG in hVSMCs-CREG cells increased compared to control cell,and the decreased obviously in hVSMCs-siCREG cells.Meanwhile,the overexpression of CREG in cells was detected to inhibit the proliferation of VSMCs and to enhance the distribution of IGF2R in cellular membrane.Furthermore,overexpression of CREG also accelerated the endocysis of IGFII in hVSMCs-CREG,and attenuate the secretion of IGFII into cell medium by ELISA analysis and Alex488 labeled IGFII analysis.Blockade experiments both neutralized antibody of IGF2R and rhIGF2R fragment determined that enhancement of IGFII secretion promoted the VSMCs proliferation,and PI3K/AKT and ERK signal pathway mediated the effect of IGFII on VSMCs.Conclusions Altogether, these data indicate that CREG inhibits the proliferation of hVSMCs through interfering into the internalization pathway of IGF2R-IGFII.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
Large conductance Ca^(2+)-activated K^+ (BK_(Ca)) channel exhibits a phenotype-dependent expression on vascular smooth muscle cells(VSMCs), which prefers to contractile phenotype. Meanwhile, shear stress definitely in...Large conductance Ca^(2+)-activated K^+ (BK_(Ca)) channel exhibits a phenotype-dependent expression on vascular smooth muscle cells(VSMCs), which prefers to contractile phenotype. Meanwhile, shear stress definitely influences VSMCs proliferation and contraction. Thereby, a hypothesis was raised, would shear stress change the BK_(Ca)expression and correlate with VSMC phenotype? In order to investigate it, VSMCs were exposed to shear stress in a parallel-plate flow chamber with 12 dynes/cm^2 for 12 h. Subsequently, the effect of shear stress on VSMC proliferation, BK_(Ca)channel expression and contractile phenotype marker, α-smooth muscle cell actin(α-SMA) and smooth muscle myosin heavy chain(SMMHC), was determined by immunofluorescence microscopy, flow cytometery as well as reverse transcriptionpolymerase chain reaction(RT-PCR), respectively. Data show that shear stress enhanced the expression of BK_(Ca)channel while inhibiting VSMC proliferation.Paralleled to those phenomena, the expression of both α-SMA and SM-MHC were decreased significantly. These results demonstrated that upregulation of BK_(Ca)channel was irrelevant to the maintenance VSMC of contractile phenotype under shear stress.This finding provides a new insight into understanding the correlation of BK_(Ca)channel and VSMC phenotype.展开更多
The effects of yam powder(YP)on the functional properties of wheat flour noodles were investigated to improve the nutritional value of noodles and provide a scientific basis for YP application in flour products.Using ...The effects of yam powder(YP)on the functional properties of wheat flour noodles were investigated to improve the nutritional value of noodles and provide a scientific basis for YP application in flour products.Using wheat flour as the main ingredient,the effects of different YP substitution ratios(0,5%,10%,15%,20%,and 25%)on polyphenolic content,2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical scavenging rate,and starch digestibility of the prepared noodles were analyzed in vitro.Additionally,the effects of YP water-soluble extracts,wheat flour noodles,and YP-enriched noodles(hereinafter,YP noodles)on inflammatory factors in vascular smooth muscle cells(VSMCs)treated by H_(2)O_(2)(100μM)were investigated.The results showed that YP had a substantial effect on polyphenolic content and DPPH free radical scavenging rate of wheat flour noodles,significantly increasing both values compared with those of the control group(P<0.05).The polyphenolic content reached a maximum of 2.17 mg/g in noodles with 20%YP substitution,whereas DPPH free radical scavenging rate reached a maximum of 42.10%in noodles with 25%YP substitution.With an increase in YP amount,both starch hydrolysis rate and rapidly digestible starch content of the noodles decreased gradually,whereas resistant starch content increased significantly.Moreover,50,100,and 200μg/mL of water-soluble YP and YP noodle extracts had no significant effect on the viability of VSMCs,and high concentrations of YP and YP noodle extracts resulted in cell death.Both YP and noodle extracts also reduced the secretion levels of inflammatory factors IL-6 and TNF-α in H_(2)O_(2)-treated VSMCs.Overall,YP addition reduced starch digestibility,increased antioxidative activity,and effectively improved functional quality of noodles.This study provides reference for the research and development of health-benefiting anti-inflammatory foods.展开更多
文摘目的:(1)观察补肾活血通络法对动脉粥样硬化易损斑块的稳定作用及其稳定斑块的可能机制;(2)探讨血清和斑块局部受激活调节正常T细胞表达和分泌因子(Regulated on Activated Normal T cell Expressed and Secreted factor,RANTES)水平与血管平滑肌细胞(vascular smooth muscle cells,VSMCs)凋亡程度的相关性。方法:将健康雄性新西兰兔34只,随机分成空白对照组(K组)和造模组(B组)。空白对照组8只,给予普通饲料喂养18周。造模组(B组)26只,给予高脂饲料喂养,实验开始第1周一次性注射牛血清白蛋白,实验第10周,随机抽取2只兔做病理检测证实模型组已形成粥样斑块。其余B组兔随机分为模型对照组、辛伐他汀组、健脑软脉方组。辛伐他汀组、健脑软脉方组分别予以辛伐他汀、健脑软脉方水煎剂干预,模型对照组予以生理盐水灌胃,持续8周,实验结束,于处死动物前24 h和48 h 2次给予药物触发易损斑块(中国斑点蝰蛇毒和组胺)。空白对照组(K组)按上述给药时间和部位给予同等剂量无菌生理盐水注射。实验终点处死实验兔,开胸迅速分离主动脉全长,检测各项指标。结果:模型对照组的主动脉病变程度、血脂水平、血清RANTES水平、斑块内RANTES m RAN和蛋白表达水平、VSMCs凋亡率与健脑软脉方组、辛伐他汀组、空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组与空白对照组相比有显著差异(P<0.05),健脑软脉方组和辛伐他汀组之间无显著差异(P>0.05)。结论:本研究表明补肾活血通络法可能通过调节血脂水平,抑制炎症反应,抑制平滑肌细胞凋亡,从而抑制模型兔动脉粥样硬化斑块的不稳定性进展。
基金supported by 2006 Great Basic Science Research Project of Jiangsu College and University(No. 06KJA36022)
文摘A new cerebroside,1-O-(β-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2′R)-2’-hydroxylignoceranoylamino]-4,8-tetradecene-3- diol was isolated from the 60%EtOH extract of traditional Chinese medical plant Cyperus rotundus L.Its structure was determined on the basis of spectroscopic data.This new compound showed anti-proliferation effect on vascular smooth muscle cells(VSMCs).
文摘Background To study the molecular mechanisms of CREG(the cellular repressor of E1A-stimulated gene) on proliferation of VSMCs in vitro.Methods The pRc/CMV-CREG plasmid or the pSM2-siCREG plasmid was transferred into human vascular smooth muscle cells(hVSMCs) to produce the cell clone that over-expression or down-expression of CREG respectively.BrdU assay and FACS cell cycle analysis were used to detect the proliferation of cells.Western blotting and immunocytochemistry show the expression and localization of IGF2R in hVSMCs.RT-PCR and ELISA assay determined the expression and secretion of IGFII factor. Alex488-labeled rhIGFII was used to investigate the endocysis of cells.And the blockade of IGFII internalization by treatment both the neutralized antibody of anti-IGF2R and rsIGF2R detected the effect of IGFII on VSMCs growth. Furthermore,Western blotting and signal pathway inhibitor were used to analysis the activation of PI3K/AKT and ERK on VSMCs proliferation.Results Western blotting identified that the expression of CREG in hVSMCs-CREG cells increased compared to control cell,and the decreased obviously in hVSMCs-siCREG cells.Meanwhile,the overexpression of CREG in cells was detected to inhibit the proliferation of VSMCs and to enhance the distribution of IGF2R in cellular membrane.Furthermore,overexpression of CREG also accelerated the endocysis of IGFII in hVSMCs-CREG,and attenuate the secretion of IGFII into cell medium by ELISA analysis and Alex488 labeled IGFII analysis.Blockade experiments both neutralized antibody of IGF2R and rhIGF2R fragment determined that enhancement of IGFII secretion promoted the VSMCs proliferation,and PI3K/AKT and ERK signal pathway mediated the effect of IGFII on VSMCs.Conclusions Altogether, these data indicate that CREG inhibits the proliferation of hVSMCs through interfering into the internalization pathway of IGF2R-IGFII.
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.
基金The National Natural Science Foundation of Chinagrant number:10972024,11120101001,10925208,and 10802006+3 种基金Doctoral Fund of Ministry of Education of Chinagrant number:0800061001National Basic Research Program of Chinagrant number:2011CB710901
文摘Large conductance Ca^(2+)-activated K^+ (BK_(Ca)) channel exhibits a phenotype-dependent expression on vascular smooth muscle cells(VSMCs), which prefers to contractile phenotype. Meanwhile, shear stress definitely influences VSMCs proliferation and contraction. Thereby, a hypothesis was raised, would shear stress change the BK_(Ca)expression and correlate with VSMC phenotype? In order to investigate it, VSMCs were exposed to shear stress in a parallel-plate flow chamber with 12 dynes/cm^2 for 12 h. Subsequently, the effect of shear stress on VSMC proliferation, BK_(Ca)channel expression and contractile phenotype marker, α-smooth muscle cell actin(α-SMA) and smooth muscle myosin heavy chain(SMMHC), was determined by immunofluorescence microscopy, flow cytometery as well as reverse transcriptionpolymerase chain reaction(RT-PCR), respectively. Data show that shear stress enhanced the expression of BK_(Ca)channel while inhibiting VSMC proliferation.Paralleled to those phenomena, the expression of both α-SMA and SM-MHC were decreased significantly. These results demonstrated that upregulation of BK_(Ca)channel was irrelevant to the maintenance VSMC of contractile phenotype under shear stress.This finding provides a new insight into understanding the correlation of BK_(Ca)channel and VSMC phenotype.
基金financial support of the Henan Provincial Science and Technology Research Project(Project No.242102310322)Shangqiu Product Quality Inspection and Testing Center(Project No.2024001).
文摘The effects of yam powder(YP)on the functional properties of wheat flour noodles were investigated to improve the nutritional value of noodles and provide a scientific basis for YP application in flour products.Using wheat flour as the main ingredient,the effects of different YP substitution ratios(0,5%,10%,15%,20%,and 25%)on polyphenolic content,2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical scavenging rate,and starch digestibility of the prepared noodles were analyzed in vitro.Additionally,the effects of YP water-soluble extracts,wheat flour noodles,and YP-enriched noodles(hereinafter,YP noodles)on inflammatory factors in vascular smooth muscle cells(VSMCs)treated by H_(2)O_(2)(100μM)were investigated.The results showed that YP had a substantial effect on polyphenolic content and DPPH free radical scavenging rate of wheat flour noodles,significantly increasing both values compared with those of the control group(P<0.05).The polyphenolic content reached a maximum of 2.17 mg/g in noodles with 20%YP substitution,whereas DPPH free radical scavenging rate reached a maximum of 42.10%in noodles with 25%YP substitution.With an increase in YP amount,both starch hydrolysis rate and rapidly digestible starch content of the noodles decreased gradually,whereas resistant starch content increased significantly.Moreover,50,100,and 200μg/mL of water-soluble YP and YP noodle extracts had no significant effect on the viability of VSMCs,and high concentrations of YP and YP noodle extracts resulted in cell death.Both YP and noodle extracts also reduced the secretion levels of inflammatory factors IL-6 and TNF-α in H_(2)O_(2)-treated VSMCs.Overall,YP addition reduced starch digestibility,increased antioxidative activity,and effectively improved functional quality of noodles.This study provides reference for the research and development of health-benefiting anti-inflammatory foods.
