【摘要】：目的研究β腎上腺素受體(adrenergic receptor, AR)-腺苷酸環(huán)化酶(adenylate cyclase, AC)-環(huán)磷酸腺苷(cyclic adenosine monophosphate,cAMP)信號通路對血管平滑肌細胞(vascular smooth muscle cell, VSMC)增殖和線粒體融合蛋白2(mitofusin2, Mfn2)及其下游信號分子表達的影響。 方法體外培養(yǎng)大鼠VSMC,給予不同濃度的(5μM、10μM、25μM、50μM)的異丙腎上腺素(isoprenaline, ISO)處理,用細胞計數(shù)、水溶性四甲基偶氮唑鹽(WST-1)法觀察ISO對VSMC增殖的影響;不同濃度(5μM、10μM、25μM、50μM)福司柯林(forskolin, FSK)處理VSMC,Western blot方法檢測兩種處理方法對Mfn2、磷酸化Raf-1(p-Raf-1)和磷酸化ERK1/2(p-ERK1/2)表達的影響;小干擾RNA(small interfering RNA , siRNA)沉默Mfn2,Western blot觀察各組Mfn2、p-ERK1/2的變化。 結(jié)果細胞計數(shù)及WST-1顯示,ISO抑制VSMC增殖,呈濃度依賴性;Western blot結(jié)果顯示,ISO及forskolin均可上調(diào)Mfn2表達,呈濃度依賴性增高,對p-Raf-1和p-ERK1/2的表達均呈濃度依賴性降低,Mfn2沉默能夠拮抗ISO介導(dǎo)的p-ERK1/2抑制作用。 結(jié)論1、細胞水平證實激動βAR能夠抑制VSMC增殖;2、βAR-AC-cAMP信號通路能夠上調(diào)Mfn2表達,通過抑制Ras-Raf-ERK/MAPK信號通路抑制VSMC增殖,表明βAR-AC-cAMP信號通路是Mfn2的一條上游信號調(diào)節(jié)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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R346

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