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1.
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   

2.
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   

3.
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   

4.
AIM: To study the effect of bFGF on cell proliferation, secretion of type I collagen and expression of integrin β1 in human kidney fibroblasts(KFB). METHODS: The KFB was cultured and stimulated by bFGF in vitro. The proliferation and collagen I secreting of KFB, the expression of integrin β1 were measured by MTT, ELISA and flow cytometer, respectively. RESULTS: bFGF(25-50 μg/L) could obviously stimulate the cell proliferation(P< 0.05), promote the secretion of collagen I(P< 0.05) and enhance the expression of integrin β1(P< 0.05) in human kidney fibroblast. CONCLUSION: bFGF could induce renal interstitial fibrosis by promoting cell proliferation, secretion of collagen I and integrin β1 expression of KFB.  相似文献   

5.
AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β-particles emission from 188Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by 188Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [3H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of 188Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from 188 Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G0/G1 arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.  相似文献   

6.
AIM: To investigate the role of α1 and β2 adrenoceptors(α1AR and β2AR) in the proliferation of hypoxic pulmonary artery smooth muscle cells (PASMCs).METHODS: PASMCs were isolated by an explant method from neonatal bovine pulmonary arteries. The cultured PASMCs were exposed to 6.6% O2 for 6 h, 12 h and 24 h. The method of -TdR incorporation was used to measure the proliferation of PASMCs. i was assayed with Fura-2/AM. The mRNA expression of α1AR, β2AR, c-fos and c-myc was determined by Northern blotting. The effects of activation of α1AR and β2AR, and inhibition of α1AR on the above indexes were observed by treating PASMCs with different AR agonists and antagonists under hypoxic condition.RESULTS: Significant increase in TdR incorporation in hypoxic PASMCs with α1AR activation was observed, and marked decrease in that was induced by α1AR inhibition. However, no significant change was found after β2AR activation. i , the mRNA expression of c-fos, c-myc, α1AR and β2AR in PASMCs were increased after hypoxia.CONCLUSION: Hypoxia induces the increase in i and mRNA expression of c-fos and c-myc, leading to the proliferation of PASMCs. The hypoxic proliferation of PASMCs is intervened by α1AR, but not β2AR. The remodeling of pulmonary arteriole and pulmonary hypertension may be involved in the processes of pulmonary arteriole constriction and proliferation induced by hypoxia through up-regulation of α1AR.  相似文献   

7.
AIM:To investigate the crosstalk between angiotensin Ⅱ (AngⅡ)-mediated and platelet-derived growth factor (PDGF)-mediated signal transduction in vascular smooth muscle proliferation.METHODS:A model of renal hypertension was made by two kidney/one-clip operation. Level of PDGF receptor β subunit of aorta was measured by Western Blot analysis. The effect of Ang Ⅱ on PDGF receptor β subunit expression was investigated in culture rat aortic vascular smooth muscle cells (VSMC).RESULTS:Systolic blood pressure obviously increased at 8th week after operation, whereas the level of PDGF receptor β subunit of aorta significantly increased by 126.6% (P＜0.05) in 2K1C rats compared with control group. The expression of PDGF receptor β subunit in cultured VSMC stimulated by AngⅡ was higher than that of control by 192.74%(P＜0.01). The effect of AngⅡ was inhibited remarkably by pretreated with losartan, a kind of specific AngⅡ receptor 1 (AT1) subtype antagonist and U73122, a kind of phospholipase C inhibitor. The effect was partly blocked by PD98059, which inhibit the activity of mitogen-activated, ERK-activating kinase (MEK).CONCLUSION:AngⅡ-induced PDGF receptor β subunit expression is regulated by the AT1 and its downstream signal molecule-PLC and ERK, might participate in the intracellular signal transduction pathway.  相似文献   

8.
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   

9.
AIM: To study the effect of basic fibroblast growth factor(bFGF) to the expression of integrin β1 subunit of endothelial cells. METHOD: The expression of integrin β1 subunit of endothelial cells was determined before and after bFGF treatment with Western Blot and immage analysis method.RESULTS: The mean gray value of immunostain by immage analysis method is 166.11±9.86 in the experimental group and 175.32±5.12 in the control group, suggested that bFGF may upregulate expression of integrin β1 subunits of endothelial cell. CONCLUSION: bFGF may play an important role in angiogenesis by way of inducing the overexpression of endothelial cell integrin β1 subunits.  相似文献   

10.
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   

11.
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   

12.
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13.
AIM: To observe the expression of Akt/GSK-3β/Snail signaling pathway-related molecules in cisplatin-resistant cell line A549/DDP mediated by transforming growth factor-β1 (TGF-β1), and to explore the association of Akt/GSK-3β/Snail signaling pathway with epithelial-mesenchymal transition (EMT). METHODS: The A549/DDP cells were divided into TGF-β1 (+) group, TGF-β1 (-) group and LY294002 group. The morphological changes of A549/DDP cells treated with TGF-β1 were observed under microscope. The protein expression of E-cadherin and N-cadherin was determined by the methods of immumofluorescence and Western blot. The protein levels of Akt, p-Akt, GSK-3β, p-GSK-3βSer9 and Snail were also detected by Western blot. RESULTS: The A549/DDP cells in TGF-β1 (+) group were dispersive, showed a spindle-like shape and developed pseudopodia. This transformation was conformed to classic EMT markers. Compared with TGF-β1 (-) group, the protein expression of E-cadherin in TGF-β1 (+) group was significantly decreased (P<0.05), and N-cadherin was significantly increased (P<0.05). The protein levels of p-Akt, p-GSK-3βSer9 and Snail were also significantly increased (P<0.05). Compared with TGF-β1 (+) group, the protein levels of p-Akt, p-GSK-3βSer9 and Snail were significantly decreased in LY294002 group (P<0.05). No difference of Akt and GSK-3β expression between TGF-β1 (-) group and TGF-β1 (+) group was observed. CONCLUSION: The mechanism of EMT in A549/DDP cells might be related to Akt/GSK-3β/Snail signaling pathway activated by TGF-β1.  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 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   

16.
AIM: To investigate the changes of endometrial receptivity under the effect of mouse embryo both in vitro and in vivo, and to figure out which part of the embryo induces the change. METHODS: Scanning electron microscope was applied to observe the pinpode formation on day 4 endometrium both in vitro and in vivo. The expression of integrin β3 and leukaemia-inhibitory factor(LIF) on day 2 pregnant mouse endometrium , day 4 endometrium after co-culture for 2 d with day 2 embryo , blastomere and zona pellucida as well as control group in vitro were detected by the methods of fluorescent quantitative PCR, immunohistochemistry and Western blotting. The same tests were conducted in the in vivo part of the experiment with the integral embryo or different parts of the embryo being transferred to pseudopregnant mouse uterus. On day 4 of the pregnancy, the endometrium was extracted to carry out the tests. RESULTS: After co-cultured for 2 d with whole embryo, the experssion of integrin β3 and LIF was higher than that in any other group in the in vitro part. The expression of integrin β3 and LIF on day 4 of normal pregnancy was higher than that in any other group in the in vivo experiments. CONCLUSION: Mouse embryo as a whole is able to induce better endometrial receptivity, while any separated part of embryo, such as blastomere or zona pellucida, couldn't.  相似文献   

17.
AIM: The effect of urotensin II (U-II) on proliferation of cultured pulmonary arterial smooth muscle cells (PASMCs) of rabbits and its mechanism are investigated. METHODS: PASMCs were isolated using explant technique. RPASMCs were incubated in serum-free medium with different concentrations of nicardipine, calcimodulin antagonist W7, PKC inhibitor H7 or MAPK inhibitor (PD98059), with or without U-II. RPASMC proliferation was examined by MTT assay and by the increase in [3H]-thymidine incorporation into DNA. RESULTS: U-II (10-9 mol/L-10-7 mol/L) increased A value of PASMCs by MTT assay and [3H]-thymidine incorporation in PASMCs in a dose-dependent manner. U-II induced a maximal effect at a concentration of 10-7 mol/L. A value and [3H]-thymidine incorporation rose 42.9% and 68.5% (P<0.05), respectively. U-II had no effect at a concentration of 10-10 mol/L. Nicardipine, W7, H7, PD98059 (10-7 mol/L-10-5 mol/L) inhibited the effect of U-II in inducing increase of A value and -thymidine incorporation in a dose-dependent manner, with the maximal inhibitory rate of 42.3%, 19.6%, 23.2%, 10.5% (P<0.05) in A value and 46.6%, 9.8%, 21.7%, 14.7% (P<0.05) in [3H]-thymidine incorporation at concentration of 10-5 mol/L, respectively. CONCLUSION: Our results suggest that U-II may induce proliferation of PASMCs of rabbits by Ca2+, CaM, PKC and MAPK signal transduction pathway.  相似文献   

18.
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   

19.
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   

20.
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   

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