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1.
AIM:To investigate the effects of fibronectin (FN) on the proliferation and collagen synthesis in cultured rat cardiac fibroblasts (CFb) derived from SHR (CFbSHR) and WKY (CFbWKY). METHODS:CFb derived from 12-week-old spontaneously hypertensive rat (SHR) and WKY was cultured by outgrowth of tissue block. Cell proliferation of CFb was measured by cell number counting and[3H]-TdR incorporation using 24-well plates pre-coated with 5 μg/cm2 of FN. Collagen synthesis was determined by [3H]-proline incorporation. RESULTS:As compared with control, the cell number of fibroblasts derived from SHR and WKY were significantly increased to 163.75% and 170.42% respectively after 72 h incubation with FN in the presence of 0.4% FCS from a intial cell density of 1×104 cells/mL. DNA synthesis of CFb was markedly promoted by FN. FN induced an increased in [3H]-proline incorporation in both CFbSHR and CFbWKY. CONCLUSION:FN is able to promote cell proliferation and collagen synthesis of CFb derived both from SHR and WKY.  相似文献   

2.
AIM:To study the effects of exogenous metallothionein (MT) and ZnCl2-induced MT production on biological action of homocysteine(HCY)in vascular fibroblasts.METHODS:[3H]-TdR, [3H]-Pro incorporation and LDH leakage were measured, the cellular viabilities were calculated by trypan blue exclusion test and the intracellular contents of MT were assayed by [109Cd]-hemoglobin saturation method in cultured rat vascular fibroblasts.RESULTS:Proliferation, collagen production of vascular fibroblasts in HCY-treated group were significantly increased compared with control group in a concentration-depedant manner. HCY (500 μmol/L) increased LDH leakage and decreased the cellular viabilities (P<0.05 or P<0.01). [3H]-TdR incorporation, [3H]-Pro incorporation, collagen secretion and LDH leakage were all decreased in MT (1×10-5 mol/L, 1×10-4mol/L) plus HCY(500 μmol/L) incubated group, compared with HCY alone group, respectively (P<0.05 or P<0.01). MT content in ZnCl2 pretreatment group was increased compared with control group. Proliferation, collagen production and LDH leakage in HCY group pretreated with ZnCl2 were decreased whereas the cellular viabilities were increased compared with HCY alone group.CONCLUSIONS:The results shows that HCY induces proliferation and collagen production of vascular fibroblasts. Both exogenous MT and endogenous MT induced by ZnCl2 inhibite the above-mentioned effects of HCY on vascular fibroblasts. MT inhibites vascular fibroblast activation induced by HCY, which may be related to its vascular protection.  相似文献   

3.
AIM: To study the effect of homocysteine (HCY) on proliferation of airway smooth muscle cells and fibroblasts and the effect of HCY on collagen prodution of airway fibroblasts. METHODS: [3H]-TdR incorpora- tion was measured in cultured airway smooth muscle cells. The [3H]-TdR and [3H]-proline incorporation were mea- sured in cultured airway fibroblasts. RESULTS: HCY induced proliferation of airway smooth muscle cells and fibroblasts in a concentration - dependent manner. HCY also induced collagen production of airway fibroblasts in a concentration - dependent manner. The inhibitors of protein kinase C, H7 and polymyxin B, inhibited HCY - induced proliferation of airway smooth muscle cells. CONCLUSIONS: HCY induced proliferation of airway smooth muscle cells and fibroblasts, HCY also induced collagen production of airway fibroblasts. The HCY - induced proliferation of airway smooth muscle cells may be related to the pathway of PKC signal transduction.  相似文献   

4.
番茄E8启动子乙烯应答元件克隆及DNA序列分析   总被引:2,自引:0,他引:2  
E8启动子是常用的番茄果实特异表达启动子之一,是指番茄E8基因5’侧翼近2.2kb的DNA序列。前人的研究表明,E8基因5’侧翼-2181~-1088区段的删除使E8基因表达量大幅度下降(仅为完整E8启动子的1/10),同时该区段还是E8基因对乙烯应答的充分必要区域;这说明了该区段是E8基因表达调控的重要元件。  相似文献   

5.
CAI Wei  XU Yi 《园艺学报》2004,20(9):1627-1630
AIM: To observe the effects of Yangxue qingnao-containing serum on rat vascular smooth muscle cell (VSMC) proliferation induced by lysophosphatidic acid (LPA). METHODS: The [3H]-TdR incorporation and mitogen-activated protein kinasc (MAPK) activity were measured in cultured VSMC. End product of lipid peroxidation-MDA levels were also detected. RESULTS: 1×10-9,1×10-8 and 1×10-7 mol/L LPA enhanced the cultured VSMC [3H]-TdR incorporation, increased MAPK activity and MDA content in a concentration-dependent manner. 5%, 10% and 15% Yangxue qingnao-containing serum concentration-dependently inhibited the increase in VSMC [3H]-TdR incorporation, MAPK activity and MDA content induced by LPA. CONCLUSIONS: LPA has a stimulating effect on VSMC proliferation. The LPA-induced intracellular signal transduction may be related to MAPK activity. Yangxue-qingnao can efficiently inhibit LPA -induced VSMC proliferation,MAPK activity and lipid peroxidation.  相似文献   

6.
AIM:To observe the effect of adrenomedullin(ADM)on proliferation of vascular smooth muscle cells(VSMC) induced by urotensin Ⅱ(UⅡ). METHODS:DNA synthesis of cultured rat aortic VSMC was measured by [3H]-TdR incorporation. The activities of mitogen activated protein kinase(MAPK) were determined by isotope tagged with [γ-32P]-ATP. RESULTS:UⅡ(10-8mol/L) significantly increased [3H]-TdR incorporation of VSMC and MAPK activities by 38%(P<0.05) and 260%(P<0.01) respectively compared with control group. Compared with UⅡ group, 10-10,10-9,10-8mol/L ADM decreased [3H]-TdR incorporation of VSMC by 7%(P>0.05), 32%(P<0.05)and 41%(P<0.01),respectively, and diminished MAPK activities by 24%(P>0.05), 32%(P<0.05)and 36%(P<0.05),respectively. CONCLUSION:ADM inhibits proliferation of VSMC induced by urotensin Ⅱ through inhibiting MAPK activation.  相似文献   

7.
AIM: To investigate the bio-effects of salusins on rat heart and cardiomyocytes. METHODS: The cardiac function was determined by multipurpose polygraph in isolated rat heart treated with various concentrations of salusin-α or salusin-β.[45Ca2+] and[3H]-Leu incorporation were determined in cultured neonatal rat cardiomyocytes with β-liquid scintillation counter. RESULTS: 10-12-10-7mol/L salusin-α and salusin-β had no effects on isolated rat cardiac function. However, salusin-α and salusin-β stimulated uptake and[3H]-Leu incorporation. The [45Ca2+] uptake induced by salusins were inhibited by nicardipine, and were synergistically increased by endothelin-1. The[3H]-Leu incorporation induced by salusin-α and salusin-β was inhibited by nicardipine, FK506 (a special inhibitor of carcineulin), PD98059 (inhibitor of MAPK) and chelerthine (inhibitor of PKC). The effects of salusin-β[45Ca2+] on uptake was stronger than those of salusin-α. But there were no statistical difference in[3H]-Leu incorporation between salusin-α and salusin-β. CONCLUSIONS: Salusin-α and salusin-β did not affect directly cardiac function in rat hearts. But salusins improved calcium uptake and protein synthesize in neonatal rat cardiomyocytes. Those effects of salusins were related with calcium channel, carcinuelin, MAPK and PKC signal pathways. Salusins may be the regulatory factors for myocardium growth and hypertrophy.  相似文献   

8.
AIM: To investigate the effects of intracellular free calcium ([Ca2+]i) from different resources on the proliferation mediated by mitogen activated protein kinase (MAPK) in vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs were used in all experiments. Calcium influx was stimulated by angiotension Ⅱ(Ang Ⅱ). The release of intracellular calcium stores was induced by inositol trisphosphate (IP3) and ryanodine (RY). MAPK activity was measured by [γ-32P]-ATP incorporation MAPK protein expression by western blot, VSMCs proliferation by [3H]-Leucine ([3H]-Leu) and [3H]-Thymidine ([3H]-TdR) incorporation. RESULTS: Compared to the control VSMCs, Ang Ⅱ, IP3 and RY significantly increased [Ca2+]i concentration activity of MAPK and its protein content in VSMCs. The promotion of [3H]-Leu and [3H]-TdR incorporation in VSMCs was also observed (P<0.01). CONCLUSION: The study indicated that calcium activator-induced increase in the activity and protein content of MAPK was involved in the proliferation of VSMCs, which was closely related to the [Ca2+]i concentration but independent to its origin.  相似文献   

9.
AIM:To explore the influence of ginsenoside Rh2 on cultured PC-3M cell cycle in vitro. METHODS:DNA content was measured using [3H]-TdR incorporation assay. To analysis the cycle changes of PC-3M cells, flow cytometry was used in the experiment.RESULTS:The results indicated that the rate of [3H]-TdR incorporation was lower in Rh2-treated cells than the control in a dose-dependent manner. Flow cytometry demonstrated that the pecent of G1 phase treated with Rh2 was higher than that of control. CONCLUSION:These results suggested that PC-3M cells cultured with Rh2 for 24 h were blocked at G1 phase, and Rh2 inhibited the growth of PC-3M cells in vitro.  相似文献   

10.
AIM: To study the effect of thyroid hormone on the expressional change of myosin heavy chain(MHC) gene in cardiomyocyte induced by angiotensinⅡ(AngⅡ) and its potential mechanism. METHODS: Cardiac myocyte was cultured according to the method of Simpson. 10-8 mol/L T3 and 10-7 mol/L AngⅡ were added to the culture medium, respectively or synchronously. After 48 h, the expression of α and β-MHC mRNA in myocytes were detected by RT-PCR. The protein kinase C activation were detected by PepTag non-radioactive PKC assay. The incorporation of -Leucine and [3H]-thymine to test the protein and DNA synthesis in myocytes were also performed. RESULTS: AngⅡalone increased the incorporation of [3H]-Leucine of myocytes while it had no effect on the incorporation of [3H]-thy mine. The expression of β-MHC mRNA was increased and the expression of α-MHC mRNA was decreased significantly at the condition of AngⅡ. The enhanced PKC activation was induced by AngⅡalso. When AngⅡand T3 were added to the culture medium synchronously, though the incorporation of [3H]-leucine and [3H]-thymine were not changed compared with AngⅡ treated alone. The α-MHC mRNA expression was increased and the β-MHC mRNA expression was decreased significantly. The PKC activation of the myocytes also was decreased. CONCLUSIONS: T3 inhibited the expressional change of myosin heavy chain gene in cardiac myocytes induced by AngⅡ. The effect of T3 on the change of PKC activation in cardiac myocytes may be one of its mechanisms.  相似文献   

11.
AIM:To observe the effect of simvastatin on the proliferation of vascular smooth muscle cells(VSMCs) induced by serum and growth factor PDGF-BB and the effect of simvastatin on the expression of PTEN,a important regulator of G1/S cell cycle transition. METHODS:The DNA synthesis was determined by [3H]-TdR incorporation, cell cycle was examined with flow cytometry, the protein level of PTEN was measured by Western blot method. RESULTS: (1)Simvastatin inhibited [3H]-TdR incorporation in a dose dependent manner. (2) Flow cytometric DNA analysis revealed that simvastatin induced significantly enhancement of G0/G1 phase and decrease in S phase VSMCs.(3)Simvastatin increased protein level of PTEN and mevalonate, a metabolite of HMG-COA, reversed the effect of simvastatin on PTEN protein expression. CONCLUSION:Simvastatin may inhibit proliferation of VSMCs and retarded cell cycle in G0/G1 phase by increasing PTEN expression through inhibiting synthesis of mevalonate.  相似文献   

12.
AIM: To investigate the effect of taurine on calcification of vascular smooth muscle cells (VSMCs).METHODS: Calcified VSMCs of rat in vitro were induced by β-glycerophosphate. Cellular calcium content, alkaline phosphatase(ALP) activities and [45Ca]accumulation were measured. DNA synthesis were evaluated by [3H]-thymidine ( [3H]-TdR) incorporation. RESULTS: Calcium content, ALP activities and [45Ca]uptake of calcified VSMCs stimulated by taurine (5-20 mmol/L) were greatly decreased in a concentration-dependent manner as compared with calcified group (P<0.01). Taurine also inhibited the proliferation of calcified cells in a concentration-dependent manner. Cell countingz, [3H]-TdR incorporation of calcified cells stimulated by taurine were greatly decreased as compared with calcified VSMCs (P<0.01). CONCLUSION: It was demonstrated that calcification of VSMCs may be alleviated by taurine.  相似文献   

13.
AIM: To investigate the effect of Salidroside on the proliferation, DNA synthesis, intracellular Ca2+ content of rabbit PASMC (pulmonary artery smooth muscle cells) under hypoxia. METHODS: Techniques of cell culture, MTT test, [3H][3H][3H]-TdR incorporation, fluo-3 and confocal laser scanning microscopy were used. RESULTS: The A value of MTT and [3H][3H]-TdR incorporation of PASMC increased significantly by 62% (P<0.05) and 138% (P<0.01) after 24 h hypoxia. Salidroside (32×10-5 mol/L) inhibited the action of hypoxia on the proliferation of PASMC, the A value of MTT and [3H][3H]-TdR incorporation declined significantly by 29% (P<0.05) and 37% (P<0.01) compared with hypoxia group. A calcium channel blocker, verapamil could also inhibit the accelerative effect of hypoxia on the proliferation of PASMC. The intracelluler Ca2+ content of PASMC raised markedly under hypoxia, but the effect of hypoxia on the intracelluler Ca2+ content could be inhibited by Salidroside. CONCLUSION: Salidroside inhibited the proliferation, DNA synthesis of PASMC induced by hypoxia. The inhibitory action of Salidroside on the increase in intracellular Ca2+ concentration under hypoxia might be one of the mechenisms.  相似文献   

14.
AIM: To observe the effect of thichosanthes injection on the expression of proliferating cell nuclear antigen (PCNA) of vascular smooth muscle cell (SMC). METHODS: The expression of PCNA of cultured rabbit aortic SMC was examined with LSAB immunohistochemical technique, and [3H]-thymidine( [3H]-TdR) incorporation data of SMC and the contents of superoxide dismutase (SOD), lipid peroxide (LPO), prostacyclin (PGI2) and cyclic AMP (cAMP) in medium were simultaneously determined. RESULTS: Thichosanthes injection has an effects of increasing SOD activity, decreasing LPO, elevating PGI2 and cAMP, reducing [3H]-TdR incorporation and expression of PCNA (all P<0.05,P<0.01). CONCLUSION: Thichosanthes could inhibit SMC proliferation.  相似文献   

15.
AIM:To investigate the effect of cyclosporine A on rat cardiomyocyte hypertrophy caused by neuropeptide Y (NPY).METHODS:Cardiomyocytes of neonatal Wistar rats were cultured with NPY or NPY together with CsA. For assessing protein synthesis rate and c-jun mRNA expression in cardiomyocytes, the methods of [3H]-Leu incorporation and RT-PCR were used.RESULTS:(1) [3H]-Leu incorporation in cardiomyocytes: [3H]-Leu incorporation in NPY (10 nmol/L) group was higher than that in control group, but there were no distinct changes between two groups. To compare with control group, [3H]-Leu incorporation in NPY (100 nmol/L) group were increased significantly (P<0.05). There was no significant change between control group and CsA group; (2) c-jun mRNA expression in cardiomyocytes: RT-PCR production of c-jun mRNA in NPY group was enhanced considerably compared with CsA group and control group (P<0.01). There was no significant change between CsA group and control group.CONCLUSIONS:NPY can induce cardiomyocyte hypertrophy. Cyclosporine A (inhibitor of CaN) can blunt the effect of NPY, suggesting that the Ca2+/CaM-dependent calcineurin (CaN) signaling pathway plays an important role in it.  相似文献   

16.
AIM: To investigate the effect of platelet-derived growth factor (PDGF) on β3 integrin gene expression and the role of β3 integrin on adhesion, migration and proliferation of vascular smooth muscle cells (VSMC) induced by PDGF. METHODS: β3 integxin gene expression was detected by RT-PCr. After β3 integrin extracellular do-main was blocks, VSMC adhesio, migration and proliferation were measured by adhesion assay awound-culture model an [3H]-TSR incorporation respectively.RESULTS: After the interaction between β3 integrin and extracellular matrix was blocked, VSMC proliferation was inhibited in some degree and the rate of [3H]-TdR incorporation into VSMC decreased 39%. The cell adhesion and migration were significantly inhibited when 10 mg/L anti-β3 integrin antibody was added (P<0.05). When VSMC were treated by PDGF for 6 hours, the expression of β3 integrin gene was 87% higher than that of control. CONCLUSION: PDGF significantly induces expression of β3 integrin gene in VSMC, and the interaction between β3 integrin and ECM protein may play an important role in VSMC adhesion and migration.  相似文献   

17.
AIM: To investigate the role of cellular paracrine in the mechanism of cardiomyocyte hypertrophy induced by stretch. METHODS: [3H]-leu incorporation into cultured cardiomyocytes was measured when stimulated by conditioned-media of myocardial fibroblast and microvascular endothelial cell after stretch. RESULTS: [3H]-leu incorporation rate were both elevated significantly stimulated by conditioned-media of myocardial fibroblast and microvascular endothelial cell. And angiotensin II and endothelin of conditioned-media of myocardial fibroblast and microvascular endothelial cell were also elevated significantly. And [3H]-leu incorporation rate were inhibited significantly when the specific angiotensin II and endothelin receptor antagonist losartan(1μmol/L) and BQ123(1μmol/L) were added. [3H]-leu incorporation rate were inhibited over 80% when losartan(1μmol/L) and BQ123(1μmol/L) were added together.CONCLUSION: The activation of myocardial fibroblast and microvascular endothelial cell endocrine stimulated by stretch play a role through paracrine in the pathogenesis of pressure-overload heart hypertrophy.  相似文献   

18.
AIM:To explore the effects of hydrogen sulfide (H2S) on proliferation of vascular smooth muscle cells (VSMC) stimulated by endothelin (ET-1, 10-7mol/L) and mitrogen-activated protein kinase (MAPK) activity in VSMCs.METHODS:Cultured VSMCs were divided into six groups: (1) control group, (2) serum group, (3) endothelin group, (4) NaHS groups, (5) serum+NaHS group, and (6) endothelin+NaHS group. VSMC proliferation was measured by[3H]-TdR incorporation and MAPK activity in VSMC was determined by radioactivity assay.RESULTS:ET-1 increased VSMC[3H]-TdR incorporation by 2.39 times (P<0.01) and MAPK activity by 1.62 times(P<0.01), as compared with control. H2S (5×10-5-5×10-4mol/L) decreased VSMC[3H]-TdR incorporation and MAPK activity by 16.8%-37.4% and 7.4%-33.6%, respectively (P<0.05 or P<0.01).CONCLUSION:This study demonstrates that H2S inhibits ET-1-induced proliferation of VSMC, which might be mediated by the inhibition of MAPK.  相似文献   

19.
AIM: To investigate the effect of caveolin-1 on the endothelin-1 (ET-1)-induced vascular smooth muscle cells (VSMC) proliferation. METHODS: The [3H]-thymidine ([3H]TdR) incorporation, immunofluorescence assays and western blotting were used in this study. RESULTS:The ETA receptor specific antagonist BQ123 inhibited the increase in[3H]TdR incorporation in response to ET-1 on VSMC.Immunofluorescence assays showed that caveolin-1 was mostly distributed in plasmalemma of VSMC.After 24 h treatment of VSMC with ET-1,the expression of caveolin-1 in VSMC was significantly decreased.Western blotting showed that ET-1 inhibited the expression of caveolin-1 in VSMC,BQ123 reversed the effect of ET-1.CONCLUSION: Caveolin-1 was mostly distributed in plasmalemma of VSMC. ET-1 downregulated caveolin-1 expression in VSMC via ETA receptor.  相似文献   

20.
AIM: To investigate the role of PI3K-IP3R-Ca2+ pathways in cardiomyocyte hypertrophy induced by tumor necrosis factor-α (TNF-α). METHODS: Myocardial cells of neonatal rats were cultured in vitro. The hypertrophic model was induced by TNF-α. The protein content was assayed with Lowry's method. The volumes of the cardiomyocytes were detected by computer photograph analysis system. The protein synthesis was determined by the method of[3H]-leucine incorporation.[Ca2+]i transient was measured by Till image system with cell-loading Fura-2/AM. RESULTS: LY294002, a PI3K inhibitor, significantly suppressed the amplitude elevation of the spontaneous[Ca2+]i transients induced by TNF-α in cultured ventricular myocytes from neonatal rats. The effect was similar to that of LY294002+2-APB (P>0.05), but lower than that in LY294002+ryanodine group (P<0.05). LY294002 significantly reduced the enhancements of protein content,[3H]-leucine incorporation and cell size induced by TNF-α. The effect was similar to that in 2-APB+LY294002 group, but higher than that in 2-APB group and lower than that in ryanodine+LY294002 group. CONCLUSION: TNF-α induces cardiac hypertrophy through PI3K-IP3R-Ca2+ pathways.  相似文献   

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