Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracell...Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs （rVSMCs） and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 （ABCA1）, adenosine triphosphate-binding cassette subfamily G member 1 （ABCG1） and peroxisome proliferator-activated receptor gamma （PPARy）. The rVSMCs were co-cultured with oxi- dized low density lipoprotein （LDL, 80 ~tg/mL） to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers （20, 40 and 60 pfu/cell） of recombinant adenovirus containing Mfn2 gene （Adv-Mfn2） were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group （P〈0.05）, and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased （P〈0.05）. At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased （P〈0.05）, but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）, and it also significantly decreased when the titer of Adv-Mfn2 increased （P〈0.05）. The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. Though the mRNA and protein expression levels of PPARy was not significantly increased （P〉0.05）, the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR＇/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.展开更多
Vascular smooth muscle cells have attracted considerable interest as a model for a flexible program of gene expression.This cell type arises throughout the embryo body plan via poorly understood signaling cascades tha...Vascular smooth muscle cells have attracted considerable interest as a model for a flexible program of gene expression.This cell type arises throughout the embryo body plan via poorly understood signaling cascades that direct the expression of transcription factors and microRNAs which,in turn,orchestrate the activation of contractile genes collectively defining this cell lineage.The discovery of myocardin and its close association with serum response factor has represented a major break-through for the molecular understanding of vascular smooth muscle cell differentiation.Retinoids have been shown to improve the outcome of vessel wall remodeling following injury and have provided further insights into the molecular circuitry that defines the vascular smooth muscle cell phenotype.This review summarizes the progress to date in each of these areas of vascular smooth muscle cell biology.展开更多
Background and Objective In-stent restenosis(ISR)remains a major limitation of percutaneous coronary intervention despite improvements in stent design and pharmacological agents,whereas the mechanism of ISR has not be...Background and Objective In-stent restenosis(ISR)remains a major limitation of percutaneous coronary intervention despite improvements in stent design and pharmacological agents,whereas the mechanism of ISR has not been fully clarified.In the present study,we sought to investigate the potential association of serum soluble TREM-1(sTREM-1)levels with the incidence of ISR.The role of TREM-1 was evaluated in cultured vascular smooth muscle cells(VSMCs).展开更多
Instruction Shear stress,caused by the parallel frictional drag force of blood flow,is a biomechanical force which plays an important role in the control of blood vessels growth and functions [1]. Clinical researches ...Instruction Shear stress,caused by the parallel frictional drag force of blood flow,is a biomechanical force which plays an important role in the control of blood vessels growth and functions [1]. Clinical researches had found out that atherosclerotic le-展开更多
Objective：ATP-binding cassette transporters（ABC） A1 and G1 play an important role in mediating cholesterol efflux and preventing macrophage foam cell formation. In this study, we examined the regulation of ABC tran...Objective：ATP-binding cassette transporters（ABC） A1 and G1 play an important role in mediating cholesterol efflux and preventing macrophage foam cell formation. In this study, we examined the regulation of ABC transporters by high glucose in human vascular smooth muscle cells（VSMCs）, the other precursor of foam cells. Methods：Incubation of human VSMCs with D-glucose（5 to 30 mM） for 1 to 7 days in the presence or absence of antioxidant and nuclear factor（NF）- κ B inhibitors, the expressions of ABCA1 and ABCG1 were analyzed by real time PCR and Western blotting. Results：High glucose decreased ABCG1 mRNA and protein expression in cultured VSMCs, whereas the expression of ABCA1 was not significantly decreased. Down-regulation of ABCG1 mRNA expression by high glucose was abolished by antioxidant N-acetyl-L-cysteine（NAC） and NF- κ B inhibitors, BAY 11-7085 and tosyl-phenylalanine chloromethyl-ketone（TPCK）. Conclusion：High glucose suppresses the expression of ABCG1 in VSMCs, which is the possible mechanism of VSMC derived foam cell transformation.展开更多
Objectives To evaluate the impact of stent implantation on proliferation and apop-tosis in injured media vascular smooth muscle cells (VSMC) and to explore the mechanism of restenosis after stent implantation. Methods...Objectives To evaluate the impact of stent implantation on proliferation and apop-tosis in injured media vascular smooth muscle cells (VSMC) and to explore the mechanism of restenosis after stent implantation. Methods Fifty male New Zealand rabbits were randomized into two groups, including balloon group and stent group. Control group was set up. The samples were harvested on 3, 7, 14, 28, 56 days after operation and the following investigation was carried out: (1) Assessing the expression of proliferating cell nuclear antigen (PCNA) of media VSMC by the method of immunohistochemistry; (2) Analyzing apoptosis of media VSMC by DNA agarose gel electrophoresis and TUNEL technique. Results The expression of PCNA and apoptosis in stent and balloon groups were markedly increased compared with control groups. (1) Stent group induced significant increased expression of PCNA in the media VSMC compared with balloon group on 3 to 28 days. On day 7, the positive rates of PCNA were 24. 36±0. 55 % vs 18. 74±1. 09 % ( P < 0. 01 ); (2) From 3 to 28 days, stent group appeared obvious DNA ladder, while balloon group only had little trace ; (3) TUNEL method showed that stent group induced much more significant apoptosis than that of balloon group on 3 to 28 days. The highest rate of apoptosis appeared on day 7: 12. 42 ±1.13% vs 5. 54±0.53% (P<0. 01); (4) By calculating the ratio of the positive rate of PCNA to apoptosis, it showed that on 3 to 28 days, the ratio of balloon group was higher than that of stent group. There was obvious difference between two groups. Conclusions Stent group induces augmented proliferation and much more significant apoptosis of media VSMC than that of balloon group. It makes the ratio of proliferation to apotosis reduced and the severity of restenosis relieved after stent implantation.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
Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultur...Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultured rat vascular smooth muscle cells （VSMCs）. Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods： Rat VSMCs were incubated with different concentrations of homocysteine （50-5000 μmol/L）. Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin （10^-9-10^-5 mol/L） were added when VSMCs were induced with 1 000 pmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results： Homocysteine （50-1000 μmol/L） increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5000 pmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and ac- tivation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine （50-5000 μmol/L） stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions： Homocysteine （50-1000 μmol/L） significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and acti- vation of MMP-2 and abnormal migration of VSMCs induced by homocysteine.展开更多
Objective： This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle ce...Objective： This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle cells （VSMCs）. Methods： Rat aortic VSMCs were isolated and cultured in vitro, and treated with different concentrations ofparthenolide （l 0, 20 and 30 μmol/L）. [^3H]thymidine incorporation was used as an index of cell proliferation. Cell cycle progression and distribution were determined by flow cytometric analysis. Furthermore, the expression of several regulatory proteins relevant to VSMC proliferation including IκBα, cyclooxygenase-2 （Cox-2）, p21, and p27 was examined to investigate the potential molecular mechanism. Results： Treatment with parthenolide significantly decreased the [^3H]thymidine incorporation into DNA by 30%-56% relative to control values in a dose-dependent manner （P〈0.05）. Addition of parthenolide also increased cell population at G0/G1 phase by 19.2%-65.7% （P〈0.05） and decreased cell population at S phase by 50.7%-84.8% （P〈0.05）, which is consistent with its stimulatory effects on p21 and p27. In addition, parthenolide also increased IκBα expression and reduced Cox-2 expression in a time-dependent manner. Conclusion： Our results show that parthenolide significantly inhibits the VSMC proliferation by inducing G0/G1 cell cycle arrest. IκBα and Cox-2 are likely involved in such inhibitory effect ofparthenolide on VSMC proliferation. These findings warrant further investigation on potential therapeutic implications ofparthenolide on VSMC proliferation in vivo.展开更多
The proliferation of vascular smooth muscle cells(VSMCs)contributes to the pathogenesis of atherosclerosis and restenosis after angioplasty and vein graft.In this study,MTT colormetry was used to test the effective sc...The proliferation of vascular smooth muscle cells(VSMCs)contributes to the pathogenesis of atherosclerosis and restenosis after angioplasty and vein graft.In this study,MTT colormetry was used to test the effective scope of emodin to inhibit VSMCs proliferation.Flow cytometry and confocal image were adopted to investigate its inhibitive mechanism.The results show that emodin could inhibit the growth and proliferation of VSMCs and the inhibition rate of emodin on VSMCs is 24.6%-94.58%,which is time-and concentration-dependent.Emodin could reduce S phase entry,increase the apoptosis of VSMCs,and reduce the intensity of[Ca^(2+)]_(i)in hPDGF B/B stimulated VSMCs.This research provides theoretical basis for medical application of emodin.It is concluded that emodin could inhibit the growth and proliferation of VSMCs effectively.Decreasing the DNA synthesis,increasing the cell apoptosis and reducing the intensity of[Ca^(2+)]_(i)in hPDGF B/B stimulated VSMCs may be the inhibitive mechanism of emodin against VSMCs proliferation.展开更多
目的研究G蛋白偶联受体APJ(血管紧张素受体样受体或称血管紧张素受体AT1相关的受体蛋白,putativere-ceptor protein related to the angiotensin receptor AT1)的内源性配体apelin-13通过PKC-ERK1/2-Cyclins信号通路促进大鼠血管平滑肌...目的研究G蛋白偶联受体APJ(血管紧张素受体样受体或称血管紧张素受体AT1相关的受体蛋白,putativere-ceptor protein related to the angiotensin receptor AT1)的内源性配体apelin-13通过PKC-ERK1/2-Cyclins信号通路促进大鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的影响。方法培养SD大鼠胸主动脉VSMCs,Western blot检测p-ERK1/2、ERK1/2、细胞周期蛋白CyclinD1和CyclinE的表达,四噻唑蓝比色法观察PKC阻断剂GF109203X对apelin-13促大鼠VSMCs增殖的影响。结果Apelin-13剂量依赖性和时间依赖性地促进大鼠VSMCsp-ERK1/2表达增加,对ERK1/2表达没有明显影响,GF109203X可明显抑制apelin-13诱导的细胞增殖及p-ERK1/2、CyclinD1和CyclinE的表达。结论Apelin-13促进大鼠VSMCs增殖可能与ape-lin-APJ-PKC-ERK1/2-Cyclins信号通路有关。展开更多
基金supported by the National Natural Science Foundation of China(No.30971244)
文摘Mitofusin2 （Mfn2） plays a pivotal role in the proliferation and apoptosis of vascular smooth muscle cells （VSMCs）. The purpose of this study was to investigate the effects of Mfn2 on the traffick- ing of intracellular cholesterol in the foam ceils derived from rat VSMCs （rVSMCs） and also to investigate the effects of Mfn2 on the expression of adenosine triphosphate-binding cassette subfamily A member 1 （ABCA1）, adenosine triphosphate-binding cassette subfamily G member 1 （ABCG1） and peroxisome proliferator-activated receptor gamma （PPARy）. The rVSMCs were co-cultured with oxi- dized low density lipoprotein （LDL, 80 ~tg/mL） to produce foam cells and cholesterol accumulation in cells. Before oxidized LDL treatment, different titers （20, 40 and 60 pfu/cell） of recombinant adenovirus containing Mfn2 gene （Adv-Mfn2） were added into the culture medium for 24 h to transfect the Mfn2 gene into the rVSMCs. Then the cells were harvested for analyses. The protein expression of Mfn2 was significantly higher in Adv-Mfn2-transfected group than in untransfected group （P〈0.05）, and the ex- pression levels significantly increased when the titer of Adv-Mfn2 increased （P〈0.05）. At 24 or 48 h af- ter oxidized LDL treatment, rVSMCs became irregular and their nuclei became larger, and their plasma abounded with red lipid droplets. However, the number of red lipid droplets was significantly decreased in Adv-Mfn2-transfected group as compared with untransfected group. At 48 h after oxidized LDL treatment, the intracellular cholesterol in rVSMCs was significantly increased （P〈0.05）, but it was sig- nificantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）, and it also significantly decreased when the titer of Adv-Mfn2 increased （P〈0.05）. The mRNA and pro- tein expression levels of ABCA1 and ABCG1 were significantly increased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. Though the mRNA and protein expression levels of PPARy was not significantly increased （P〉0.05）, the phosporylation levels of PPARy were signifi- cantly decreased in Adv-Mfn2-transfected group as compared with untransfected group （P〈0.05）. These results suggest that the transfection of Adv-Mfn2 can significantly reduce intracellular cholesterol in oxidized LDL-induced rVSMCs possibly by decreasing PPAR＇/phosporylation and then increasing pro- tein expression levels of ABCAI and ABCG1, which may be helpful to suppress the formation of foam cells.
文摘Vascular smooth muscle cells have attracted considerable interest as a model for a flexible program of gene expression.This cell type arises throughout the embryo body plan via poorly understood signaling cascades that direct the expression of transcription factors and microRNAs which,in turn,orchestrate the activation of contractile genes collectively defining this cell lineage.The discovery of myocardin and its close association with serum response factor has represented a major break-through for the molecular understanding of vascular smooth muscle cell differentiation.Retinoids have been shown to improve the outcome of vessel wall remodeling following injury and have provided further insights into the molecular circuitry that defines the vascular smooth muscle cell phenotype.This review summarizes the progress to date in each of these areas of vascular smooth muscle cell biology.
文摘Background and Objective In-stent restenosis(ISR)remains a major limitation of percutaneous coronary intervention despite improvements in stent design and pharmacological agents,whereas the mechanism of ISR has not been fully clarified.In the present study,we sought to investigate the potential association of serum soluble TREM-1(sTREM-1)levels with the incidence of ISR.The role of TREM-1 was evaluated in cultured vascular smooth muscle cells(VSMCs).
基金supported by grants from the National Natural Science Foundation of China,Nos10732070,10702043,30970703,10972140 and 30470432
文摘Instruction Shear stress,caused by the parallel frictional drag force of blood flow,is a biomechanical force which plays an important role in the control of blood vessels growth and functions [1]. Clinical researches had found out that atherosclerotic le-
基金supported by the National Natural Science Foundation of China(No.30570732),NCET-04-197"985 Project"of Ministry of Education,Study on molecular mechanism of vascular-related diseases and on gene function.
文摘Objective：ATP-binding cassette transporters（ABC） A1 and G1 play an important role in mediating cholesterol efflux and preventing macrophage foam cell formation. In this study, we examined the regulation of ABC transporters by high glucose in human vascular smooth muscle cells（VSMCs）, the other precursor of foam cells. Methods：Incubation of human VSMCs with D-glucose（5 to 30 mM） for 1 to 7 days in the presence or absence of antioxidant and nuclear factor（NF）- κ B inhibitors, the expressions of ABCA1 and ABCG1 were analyzed by real time PCR and Western blotting. Results：High glucose decreased ABCG1 mRNA and protein expression in cultured VSMCs, whereas the expression of ABCA1 was not significantly decreased. Down-regulation of ABCG1 mRNA expression by high glucose was abolished by antioxidant N-acetyl-L-cysteine（NAC） and NF- κ B inhibitors, BAY 11-7085 and tosyl-phenylalanine chloromethyl-ketone（TPCK）. Conclusion：High glucose suppresses the expression of ABCG1 in VSMCs, which is the possible mechanism of VSMC derived foam cell transformation.
文摘Objectives To evaluate the impact of stent implantation on proliferation and apop-tosis in injured media vascular smooth muscle cells (VSMC) and to explore the mechanism of restenosis after stent implantation. Methods Fifty male New Zealand rabbits were randomized into two groups, including balloon group and stent group. Control group was set up. The samples were harvested on 3, 7, 14, 28, 56 days after operation and the following investigation was carried out: (1) Assessing the expression of proliferating cell nuclear antigen (PCNA) of media VSMC by the method of immunohistochemistry; (2) Analyzing apoptosis of media VSMC by DNA agarose gel electrophoresis and TUNEL technique. Results The expression of PCNA and apoptosis in stent and balloon groups were markedly increased compared with control groups. (1) Stent group induced significant increased expression of PCNA in the media VSMC compared with balloon group on 3 to 28 days. On day 7, the positive rates of PCNA were 24. 36±0. 55 % vs 18. 74±1. 09 % ( P < 0. 01 ); (2) From 3 to 28 days, stent group appeared obvious DNA ladder, while balloon group only had little trace ; (3) TUNEL method showed that stent group induced much more significant apoptosis than that of balloon group on 3 to 28 days. The highest rate of apoptosis appeared on day 7: 12. 42 ±1.13% vs 5. 54±0.53% (P<0. 01); (4) By calculating the ratio of the positive rate of PCNA to apoptosis, it showed that on 3 to 28 days, the ratio of balloon group was higher than that of stent group. There was obvious difference between two groups. Conclusions Stent group induces augmented proliferation and much more significant apoptosis of media VSMC than that of balloon group. It makes the ratio of proliferation to apotosis reduced and the severity of restenosis relieved after stent implantation.
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).
基金Project supported by the Health Ministry Scientific Research Fund of China (No. WKJ2011-2-018)the Zhejiang Provincial Natural Science Foundation of China (No. Y2100535)+3 种基金the Key Social Development Project of Zhejiang Province (No. 2010A23010)the Science and Technology Projects of Shaoxing (No. 2011A23011)the Science and Technology Plan Project of Zhejiang Province (No. 2012C33040)the Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents, China
文摘Objective： To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 （MMP-2） and tissue inhibitors of matrix metalloproteinase-2 （TIMP-2） and on cell migration of cultured rat vascular smooth muscle cells （VSMCs）. Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods： Rat VSMCs were incubated with different concentrations of homocysteine （50-5000 μmol/L）. Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin （10^-9-10^-5 mol/L） were added when VSMCs were induced with 1 000 pmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results： Homocysteine （50-1000 μmol/L） increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5000 pmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and ac- tivation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine （50-5000 μmol/L） stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions： Homocysteine （50-1000 μmol/L） significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and acti- vation of MMP-2 and abnormal migration of VSMCs induced by homocysteine.
基金Project (No. 491020-W50315) supported by the Foundation of the Health Bureau of Zhejiang, China
文摘Objective： This study is to determine the effect of the natural product parthenolide, a sesquiterpene lactone isolated from extracts of the herb Tanacetum parthenium, on the proliferation of vascular smooth muscle cells （VSMCs）. Methods： Rat aortic VSMCs were isolated and cultured in vitro, and treated with different concentrations ofparthenolide （l 0, 20 and 30 μmol/L）. [^3H]thymidine incorporation was used as an index of cell proliferation. Cell cycle progression and distribution were determined by flow cytometric analysis. Furthermore, the expression of several regulatory proteins relevant to VSMC proliferation including IκBα, cyclooxygenase-2 （Cox-2）, p21, and p27 was examined to investigate the potential molecular mechanism. Results： Treatment with parthenolide significantly decreased the [^3H]thymidine incorporation into DNA by 30%-56% relative to control values in a dose-dependent manner （P〈0.05）. Addition of parthenolide also increased cell population at G0/G1 phase by 19.2%-65.7% （P〈0.05） and decreased cell population at S phase by 50.7%-84.8% （P〈0.05）, which is consistent with its stimulatory effects on p21 and p27. In addition, parthenolide also increased IκBα expression and reduced Cox-2 expression in a time-dependent manner. Conclusion： Our results show that parthenolide significantly inhibits the VSMC proliferation by inducing G0/G1 cell cycle arrest. IκBα and Cox-2 are likely involved in such inhibitory effect ofparthenolide on VSMC proliferation. These findings warrant further investigation on potential therapeutic implications ofparthenolide on VSMC proliferation in vivo.
文摘The proliferation of vascular smooth muscle cells(VSMCs)contributes to the pathogenesis of atherosclerosis and restenosis after angioplasty and vein graft.In this study,MTT colormetry was used to test the effective scope of emodin to inhibit VSMCs proliferation.Flow cytometry and confocal image were adopted to investigate its inhibitive mechanism.The results show that emodin could inhibit the growth and proliferation of VSMCs and the inhibition rate of emodin on VSMCs is 24.6%-94.58%,which is time-and concentration-dependent.Emodin could reduce S phase entry,increase the apoptosis of VSMCs,and reduce the intensity of[Ca^(2+)]_(i)in hPDGF B/B stimulated VSMCs.This research provides theoretical basis for medical application of emodin.It is concluded that emodin could inhibit the growth and proliferation of VSMCs effectively.Decreasing the DNA synthesis,increasing the cell apoptosis and reducing the intensity of[Ca^(2+)]_(i)in hPDGF B/B stimulated VSMCs may be the inhibitive mechanism of emodin against VSMCs proliferation.
文摘目的研究G蛋白偶联受体APJ(血管紧张素受体样受体或称血管紧张素受体AT1相关的受体蛋白,putativere-ceptor protein related to the angiotensin receptor AT1)的内源性配体apelin-13通过PKC-ERK1/2-Cyclins信号通路促进大鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)增殖的影响。方法培养SD大鼠胸主动脉VSMCs,Western blot检测p-ERK1/2、ERK1/2、细胞周期蛋白CyclinD1和CyclinE的表达,四噻唑蓝比色法观察PKC阻断剂GF109203X对apelin-13促大鼠VSMCs增殖的影响。结果Apelin-13剂量依赖性和时间依赖性地促进大鼠VSMCsp-ERK1/2表达增加,对ERK1/2表达没有明显影响,GF109203X可明显抑制apelin-13诱导的细胞增殖及p-ERK1/2、CyclinD1和CyclinE的表达。结论Apelin-13促进大鼠VSMCs增殖可能与ape-lin-APJ-PKC-ERK1/2-Cyclins信号通路有关。
