目的研究絲裂原活化蛋白激酶亞單位細(xì)胞外信號(hào)調(diào)節(jié)激酶(ERK)和p38蛋白激酶(p38 MAPK)在移植靜脈血管重塑過程中的表達(dá)。方法選Wistar大鼠80只,建立自體移植靜脈模型,術(shù)后隨機(jī)分為6 h、24 h、3 d、7 d、2周、4周、6周及8周組,于相應(yīng)時(shí)點(diǎn)取材,半定量逆轉(zhuǎn)錄PCR法檢測移植血管中ERK和p38 MAPK的mRNA表達(dá); Western蛋白印跡定量檢測ERK和p38 MAPK的蛋白產(chǎn)物及磷酸化蛋白產(chǎn)物表達(dá); 脫氧核苷酸末端轉(zhuǎn)移酶末端標(biāo)記法(TUNEL)檢測血管平滑肌細(xì)胞(VSMC)凋亡的變化; 免疫組化檢測增殖細(xì)胞核抗原(PCNA)的表達(dá)。 結(jié)果移植靜脈術(shù)后6 h,ERK1和p38 MAPK的mRNA表達(dá)均明顯增強(qiáng),與正常靜脈組比較差異均有顯著性意義(P<0.01); ERK1 mRNA表達(dá)在移植后7 d達(dá)高峰,表達(dá)值為(33.2±14.2)%,p38 MAPK的mRNA表達(dá)于術(shù)后2周達(dá)到高峰,表達(dá)值為(58.8±26.2)%,與其余各時(shí)點(diǎn)比較差異有顯著性意義(P<0.01)。Western蛋白印跡提示ERK1/2在術(shù)后1~2周達(dá)高峰,6周時(shí)逐漸恢復(fù)至正常水平; 而p38 MAPK則在移植后2~4周達(dá)高峰,之后開始減少,8周時(shí)仍維持一定表達(dá)量(1/4~1/2)。ERK1與PCNA表達(dá)呈正相關(guān)(r=0.759 6,P<0.01),p38 MAPK與凋亡呈正相關(guān)(r=0.892 2,P<0.01)。結(jié)論MAPK的激活是移植靜脈內(nèi)膜增生以及血管重塑的關(guān)鍵環(huán)節(jié),可能成為防治移植靜脈狹窄、閉塞的新的治療靶點(diǎn)。
引用本文: 楊軍,胡新華,張強(qiáng),段志泉. 細(xì)胞外信號(hào)調(diào)節(jié)激酶和p38蛋白激酶在自體移植靜脈中的表達(dá). 中國普外基礎(chǔ)與臨床雜志, 2004, 11(4): 291-294. doi: 復(fù)制
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- 2. 胡新華,張強(qiáng),孫達(dá)欣,等. 核轉(zhuǎn)錄因子κB及其抑制基因在自體移植靜脈中的表達(dá) [J]. 中華醫(yī)學(xué)雜志, 2002; 82(22)∶1546.
- 3. Sambrook J, Fritsch EF, Maniatis T. 克隆化基因所表達(dá)蛋白質(zhì)的檢測與分析 [A]. 金冬雁,黎孟楓,譯. 分子克隆實(shí)驗(yàn)指南 [M]. 第2版. 北京: 科學(xué)出版社, 1992∶852~898.
- 4. Talmor D, Applebaum A, Rudich A, et al. Activation of mitogenactivated protein kinases in human heart during cardiopulmonary bypass [J]. Circ Res, 2000; 86(9)∶1004.
- 5. Thury A, van Langenhove G, Carlier SG, et al. High shear stress after successful balloon angioplasty is associated with restenosis and target lesion revascularization [J]. Am Heart J, 2002; 144(1)∶136.
- 6. Xi XP, Graf K, Goetze S,et al. Central role of the MAPK pathway in ang IImediated DNA synthesis and migration in rat vascular smooth muscle cells [J]. Arterioscler Thromb Vasc Biol, 1999; 19(1)∶73.
- 7. Xu Q, Liu Y, Gorospe M, et al. Acute hypertension activates mitogenactivated protein kinases in arterial wall [J]. J Clin Invest, 1996; 97(2)∶508.
- 8. Ohashi N, Matsumori A, Furukawa Y, et al. Role of p38 mitogenactivated protein kinase in neointimal hyperplasia after vascular injury [J]. Arterioscler Thromb Vasc Biol, 2000; 20(12)∶2521.