血管紧张素Ⅱ对大鼠血管外膜成纤维细胞骨桥蛋白表达的影响
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Effects of Angiotensin Ⅱ on the Expression of Osteopontin in Rat Vascular Adventitial Fibroblasts
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    摘要:

    目的研究血管紧张素Ⅱ对培养的大鼠胸主动脉外膜成纤维细胞骨桥蛋白表达的影响。方法大鼠外膜成纤维细胞被随机分为对照组、血管紧张素Ⅱ不同浓度(10-8、10-7、10-6和10-5mol/L)和时间(6h、12h和24h)干预组及血管紧张素Ⅱ加AT1受体拮抗剂氯沙坦和/或AT2受体拮抗剂PD123319干预组,用RT-PCR及Western blotting技术结合光密度扫描分析,观察血管紧张素Ⅱ对培养的大鼠胸主动脉外膜成纤维细胞骨桥蛋白表达的影响。结果血管紧张素Ⅱ10-6mol/L明显刺激外膜成纤维细胞骨桥蛋白的表达,6h、12h和24h桥蛋白较对照组分别升高65.43%、97.03%和105.11%(P<0.05);而24h阴性对照无明显变化。不同浓度的血管紧张素Ⅱ均可诱导骨桥蛋白表达,10-8、10-7、10-6和10-5mol/L管紧张素Ⅱ孵育后分别升高了70.46%、97.37%、123.10%和147.59%(P<0.01)。在血管紧张素Ⅱ刺激前1h分别加入氯沙坦(10-5mol/L)、PD123319(10mmol/L)、氯沙坦(10-5mol/L)+PD123319(10mmol/L),共同孵育12h后,其抑制率分别为58.9%、0.92%和59.7%。Western blotting分析,12h不同剂量血管紧张素Ⅱ干预组(10-8、10-7、10-6和10-5mol/L)骨桥蛋白量较对照组分别提高了50.68%、63.03%和69.86%(P<0.05),氯沙坦明显抑制血管紧张素Ⅱ刺激的骨桥蛋白表达,抑制率达到61.7%(P<0.05),而PD123319作用后抑制作用不明显,抑制率仅为0.85%(P>0.05)。结论血管紧张素Ⅱ能在基因和蛋白水平上刺激大鼠外膜成纤维细胞骨桥蛋白的表达,此作用可能是通过AT1受体介导。

    Abstract:

    Aim To explore the function of angiotensin Ⅱ (AngⅡ) in promoting osteopontin gene expression and protein synthsis of rat vascular adventitial fibroblasts. Methods Cultured cells were incubated with 1 different concentration of AngⅡ for different hours and AngⅡ plus AT1 receptor blocker,losartan or/and AT2 receptor blocker,PD123319. Then RT-PCR and WesternBlotting analysis were used to observe the effect of AngⅡ on osteopontin gene expression of rat vascular adventitial fibroblasts. Results AngⅡ(10-6 mol/L)obviously enhanced the expression of OPN mRNA in adventitial fibroblasts in time-dependent manner, the ratio of mRNA of osteopontin and β-actin in cells incubated with AngⅡ for 6,12 and 24 h increased by 65.43%, 97.03% and 105.11% respectively,as compared with that of the control (P<0.05), whereas there was no significant change in negative control at 24 h. AngⅡ could induce gene expression of osteopontin in dose-dependent manner, cells incubated with 10-8, 10-7, 10-6 and 10-5 mol/L AngⅡ for 12 h, there was up-regulation of OPN mRNA expression by 70.46%, 97.37%, 123.10% and 147.59% respectively (P<0.01).Pre-treatment with losartan(10-5 mol/L) and/or PD123319(10 mmol/L) for 1h in adventitial fibroblasts before AngⅡ(10-6 mol/L) used,the expression of OPN mRNA was inhibited by 58.9%, 0.92% and 59.7% respectively . Westernblotting analysis also showed that protein synthesis of osteopontin in cells incubated with 10-8, 10-7, 10-6 and 10-5 mol/L AngⅡ for 12h increased by 50.68%, 63.03%, 69.86% respectively ,as compared with that of the control(P<0.05). Pre-treatment with losartan(10-5 mol/L) but not PD123319 (10 mmol/L) before AngⅡ (10-6 mol/L) used, could inhibited the expression of OPN protein. Conclusions AngⅡ may up-regulate osteopontin gene expression and protein synthesis of rat vascular adventitial fibroblasts via angiotensin type 1 receptor.

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刘海潮,何军,马业新.血管紧张素Ⅱ对大鼠血管外膜成纤维细胞骨桥蛋白表达的影响[J].中国动脉硬化杂志,2008,16(12):961~964.

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  • 收稿日期:2008-06-16
  • 最后修改日期:2008-11-15
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