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目的 :研究白细胞介素-17(IL-17)对大鼠肾脏成纤维细胞(NRK-49F)转化为肌成纤维细胞、表达细胞外基质的影响,并进一步探讨该调节作用可能涉及的分子机制。方法:以转化生长因子β(TGF-β)诱导培养NRK-49F细胞,采用Western blot检测IL-17对NRK-49F表达α-平滑肌肌动蛋白(α-SMA)和细胞外基质Ⅰ型胶原和纤连蛋白的影响;并运用real-time PCR分析NRK-49F在诱导培养24、48、72 h后细胞外基质Ⅰ型胶原和纤连蛋白m RNA的表达水平及IL-17的调节作用。最后,运用Western blot检测细胞内Smad与非Smad通路中相关信号分子的表达及其磷酸化水平,以解释IL-17对NRK-49F调节作用的分子机制。结果:IL-17抑制NRK-49F细胞表达α-SMA,并抑制其表达细胞外基质Ⅰ型胶原和纤连蛋白。当IL-17对NRK-49F发挥上述抑制作用时,并未影响经典Smad信号分子的表达,而是通过抑制非Smad通路Akt-TSC2-p70S6K的活化来实现的。结论:IL-17通过抑制TGF-β的非Smad通路Akt-TSC2-p70S6K的活化,从而抑制NRK-49F转化为肌成纤维细胞,并抑制其表达细胞外基质。
Objective: To investigate the effects of interleukin-17 (IL-17) on the conversion of rat renal fibroblasts (NRK-49F) to myofibroblasts and the expression of extracellular matrix (ECM), and to explore the possible molecular mechanisms involved in this regulation . Methods: NRK-49F cells were induced by transforming growth factor-β (TGF-β), and the effect of IL-17 on the expression of α-smooth muscle actin (α-SMA) and extracellular matrix type I collagen Fibronectin. The expression of type I collagen and fibronectin m RNA of NRK-49F and the regulatory effect of IL-17 on NRK-49F cells after 24, 48 and 72 h induction were analyzed by real-time PCR. Finally, Western blot was used to detect the expression and phosphorylation of related signal molecules in Smad and non-Smad pathways to explain the molecular mechanism of IL-17 on NRK-49F. RESULTS: IL-17 inhibited the expression of α-SMA in NRK-49F cells and inhibited the expression of extracellular matrix type I collagen and fibronectin. When IL-17 exerted the above inhibition on NRK-49F, it did not affect the expression of classical Smad signaling molecules, but inhibited the activation of Akt-TSC2-p70S6K, a non-Smad pathway. CONCLUSION: IL-17 inhibits the expression of extracellular matrix by inhibiting the activation of Akt-TSC2-p70S6K, a non-Smad pathway of TGF-β, thereby inhibiting the conversion of NRK-49F into myofibroblasts.