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[目的]探讨N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸(Ac-SDKP)是否通过抑制热休克蛋白27(HSP27)的表达,而抑制转化生长因子1(TGF-β1)诱导的人肺泡Ⅱ型上皮细胞向肌成纤维细胞的分化以及Ⅰ型、Ⅲ型胶原蛋白的表达。[方法]用5 ng/m L的TGF-β1诱导人肺泡Ⅱ型上皮细胞株A549 72 h,并分为对照组、TGF-β1诱导组和10-8 mol/L Ac-SDKP干预组。采用倒置相差显微镜观察A549细胞上皮-间质转化中细胞形态变化;免疫细胞化学法检测角蛋白(CK8)、波形蛋白的定位;Western blot法检测E-钙黏蛋白(E-cad)、CK8、波形蛋白、α-平滑肌肌动蛋白(α-SMA)及HSP27以及Ⅰ型、Ⅲ型胶原蛋白表达。[结果]在TGF-β1的诱导下,A549细胞向肌成纤维细胞的形态转变,伴随着上皮标志物CK8、E-cad降低,分别是对照组的40%和50%,而间质细胞标志物波形蛋白、α-SMA表达增强,同时伴随HSP27及Ⅰ型、Ⅲ型胶原蛋白表达增强,分别是对照组的1.9倍、1.8倍、1.9倍以及2.2倍、2.5倍(P<0.05)。给予Ac-SDKP干预后,CK8、E-cad表达上调,分别是TGF-β1的诱导组的2.6倍、2.0倍;而波形蛋白、α-SMA、HSP27及Ⅰ型、Ⅲ型胶原蛋白表达明显降低,分别是TGF-β1的诱导组的68%、66%、74%以及66%、44%(P<0.05)。[结论]Ac-SDKP能够通过对HSP27表达的调节,而抑制肺泡Ⅱ型上皮细胞向肌成纤维细胞的转化及胶原蛋白的合成。
[Objective] To investigate whether Ac-SDKP inhibits the expression of heat shock protein 27 (HSP27) and inhibits the expression of transforming growth factor 1 ( TGF-β1) induces the differentiation of human alveolar type Ⅱ epithelial cells into myofibroblasts and the expression of type Ⅰ and type Ⅲ collagen. [Method] Human lung alveolar type Ⅱ epithelial cell line A549 was induced with 5 ng / mL TGF-β1 for 72 h and divided into control group, TGF-β1 induction group and 10-8 mol / L Ac-SDKP intervention group. The morphological changes of epithelial-mesenchymal transition of A549 cells were observed by inverted phase contrast microscope. The localization of cytokeratin (CK8) and vimentin was detected by immunocytochemistry. The expressions of E-cadherin, CK8, Vimentin, α-smooth muscle actin (α-SMA) and HSP27 and type Ⅰ and type Ⅲ collagen protein expression. [Results] The morphological changes of A549 cells to myofibroblasts induced by TGF-β1 were accompanied by the decrease of epithelial markers CK8 and E-cad, which were 40% and 50% of the control group, respectively. The stromal cell markers The expression of α-SMA and vimentin increased with the increase of HSP27 and type Ⅰ and type Ⅲ collagen, which were 1.9 times, 1.8 times, 1.9 times and 2.2 times, 2.5 times (P <0.05) respectively. After Ac-SDKP intervention, the expression of CK8 and E-cad was up-regulated by 2.6-fold and 2.0-fold compared with that induced by TGF-β1, while the expressions of vimentin, α-SMA, HSP27 and type I and type III collagen were significantly decreased , Which were 68%, 66%, 74% and 66%, 44% (P <0.05) respectively for the induced group of TGF-β1. [Conclusion] Ac-SDKP could inhibit the transformation of alveolar type Ⅱ epithelial cells into myofibroblasts and the synthesis of collagen through the regulation of HSP27 expression.