βAR-cAMP通路通过上调Mfn2抑制血管平滑肌细胞增殖
发布时间：2018-11-18 07:25
【摘要】：目的研究β肾上腺素受体(adrenergic receptor, AR)-腺苷酸环化酶(adenylate cyclase, AC)-环磷酸腺苷(cyclic adenosine monophosphate,cAMP)信号通路对血管平滑肌细胞(vascular smooth muscle cell, VSMC)增殖和线粒体融合蛋白2(mitofusin2, Mfn2)及其下游信号分子表达的影响。 方法体外培养大鼠VSMC,给予不同浓度的(5μM、10μM、25μM、50μM)的异丙肾上腺素(isoprenaline, ISO)处理,用细胞计数、水溶性四甲基偶氮唑盐(WST-1)法观察ISO对VSMC增殖的影响;不同浓度(5μM、10μM、25μM、50μM)福司柯林(forskolin, FSK)处理VSMC,Western blot方法检测两种处理方法对Mfn2、磷酸化Raf-1(p-Raf-1)和磷酸化ERK1/2(p-ERK1/2)表达的影响;小干扰RNA(small interfering RNA , siRNA)沉默Mfn2,Western blot观察各组Mfn2、p-ERK1/2的变化。 结果细胞计数及WST-1显示,ISO抑制VSMC增殖,呈浓度依赖性;Western blot结果显示,ISO及forskolin均可上调Mfn2表达,呈浓度依赖性增高,对p-Raf-1和p-ERK1/2的表达均呈浓度依赖性降低,Mfn2沉默能够拮抗ISO介导的p-ERK1/2抑制作用。 结论1、细胞水平证实激动βAR能够抑制VSMC增殖;2、βAR-AC-cAMP信号通路能够上调Mfn2表达,通过抑制Ras-Raf-ERK/MAPK信号通路抑制VSMC增殖,表明βAR-AC-cAMP信号通路是Mfn2的一条上游信号调节通路。
[Abstract]:Objective to study the effect of 尾 -adrenergic receptor (adrenergic receptor, AR) adenylate cyclase (adenylate cyclase, AC) cyclic adenosine phosphate (cyclic adenosine monophosphate,cAMP) signal pathway on (vascular smooth muscle cell, VSMC) proliferation and mitochondrial fusion protein 2 (mitofusin2,) in vascular smooth muscle cells. Mfn2) and its downstream signal molecules. Methods Rat VSMC, was treated with isoprenaline (isoprenaline, ISO) (5 渭 M 10 渭 M, 25 渭 M, 50 渭 M) in vitro. The effect of ISO on the proliferation of VSMC was observed by cell count and water-soluble tetramethyl azolium salt (WST-1) method. The effects of different concentrations (5 渭 M, 10 渭 M, 25 渭 M, 50 渭 M) on the expression of Mfn2, phosphorylated Raf-1 (p-Raf-1) and phosphorylated ERK1/2 (p-ERK1/2) were detected by Foscolin (forskolin, FSK) treatment with VSMC,Western blot. Small interfering RNA (small interfering RNA, siRNA) silenced Mfn2,Western blot to observe the changes of Mfn2,p-ERK1/2 in each group. Results Cell count and WST-1 showed that ISO inhibited the proliferation of VSMC in a concentration-dependent manner. Western blot results showed that both ISO and forskolin up-regulated the expression of Mfn2 in a concentration-dependent manner, and decreased the expression of p-Raf-1 and p-ERK1/2 in a concentration-dependent manner. Mfn2 silencing could antagonize ISO mediated p-ERK1/2 inhibition. Conclusion 1. Activation of 尾 AR can inhibit the proliferation of VSMC at cell level. 2. 尾 AR-AC-cAMP signaling pathway can up-regulate Mfn2 expression and inhibit VSMC proliferation by inhibiting Ras-Raf-ERK/MAPK signaling pathway, which indicates that 尾 AR-AC-cAMP signaling pathway is an upstream signal regulation pathway of Mfn2.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R346
本文编号：2339309
[Abstract]:Objective to study the effect of 尾 -adrenergic receptor (adrenergic receptor, AR) adenylate cyclase (adenylate cyclase, AC) cyclic adenosine phosphate (cyclic adenosine monophosphate,cAMP) signal pathway on (vascular smooth muscle cell, VSMC) proliferation and mitochondrial fusion protein 2 (mitofusin2,) in vascular smooth muscle cells. Mfn2) and its downstream signal molecules. Methods Rat VSMC, was treated with isoprenaline (isoprenaline, ISO) (5 渭 M 10 渭 M, 25 渭 M, 50 渭 M) in vitro. The effect of ISO on the proliferation of VSMC was observed by cell count and water-soluble tetramethyl azolium salt (WST-1) method. The effects of different concentrations (5 渭 M, 10 渭 M, 25 渭 M, 50 渭 M) on the expression of Mfn2, phosphorylated Raf-1 (p-Raf-1) and phosphorylated ERK1/2 (p-ERK1/2) were detected by Foscolin (forskolin, FSK) treatment with VSMC,Western blot. Small interfering RNA (small interfering RNA, siRNA) silenced Mfn2,Western blot to observe the changes of Mfn2,p-ERK1/2 in each group. Results Cell count and WST-1 showed that ISO inhibited the proliferation of VSMC in a concentration-dependent manner. Western blot results showed that both ISO and forskolin up-regulated the expression of Mfn2 in a concentration-dependent manner, and decreased the expression of p-Raf-1 and p-ERK1/2 in a concentration-dependent manner. Mfn2 silencing could antagonize ISO mediated p-ERK1/2 inhibition. Conclusion 1. Activation of 尾 AR can inhibit the proliferation of VSMC at cell level. 2. 尾 AR-AC-cAMP signaling pathway can up-regulate Mfn2 expression and inhibit VSMC proliferation by inhibiting Ras-Raf-ERK/MAPK signaling pathway, which indicates that 尾 AR-AC-cAMP signaling pathway is an upstream signal regulation pathway of Mfn2.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R346
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