邓海静, 李世峰, 孙月, 张丽娟, 薛新新, 杜世璞, 徐洪, 杨方. Ac-SDKP在抑制人肺泡Ⅱ型上皮细胞向肌成纤维细胞分化中的作用[J]. 环境与职业医学, 2015, 32(2): 113-117. DOI: 10.13213/j.cnki.jeom.2015.14507
引用本文: 邓海静, 李世峰, 孙月, 张丽娟, 薛新新, 杜世璞, 徐洪, 杨方. Ac-SDKP在抑制人肺泡Ⅱ型上皮细胞向肌成纤维细胞分化中的作用[J]. 环境与职业医学, 2015, 32(2): 113-117. DOI: 10.13213/j.cnki.jeom.2015.14507
DENG Hai-jing , LI Shi-feng , SUN Yue , ZHANG Li-juan , XUE Xin-xin , DU Shi-pu , XU Hong , YANG Fang . Effects of Ac-SDKP on Epithelial-Mesenchymal Transition of Human Type Ⅱ Alveolar Epithelial Cells[J]. Journal of Environmental and Occupational Medicine, 2015, 32(2): 113-117. DOI: 10.13213/j.cnki.jeom.2015.14507
Citation: DENG Hai-jing , LI Shi-feng , SUN Yue , ZHANG Li-juan , XUE Xin-xin , DU Shi-pu , XU Hong , YANG Fang . Effects of Ac-SDKP on Epithelial-Mesenchymal Transition of Human Type Ⅱ Alveolar Epithelial Cells[J]. Journal of Environmental and Occupational Medicine, 2015, 32(2): 113-117. DOI: 10.13213/j.cnki.jeom.2015.14507

Ac-SDKP在抑制人肺泡Ⅱ型上皮细胞向肌成纤维细胞分化中的作用

Effects of Ac-SDKP on Epithelial-Mesenchymal Transition of Human Type Ⅱ Alveolar Epithelial Cells

  • 摘要: 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/mL的TGF-β1诱导人肺泡Ⅱ型上皮细胞株A54972 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

     

    Abstract: Objective To investigate whether N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) could inhibit epithelialmesenchymal transition of human typeⅡ alveolar epithelial cells through suppressing the expression of heat shock protein27 (HSP27) and the deposition of typeⅠand type Ⅲ collagen mediated by transforming growth factor beta 1 (TGF-β1).

    Methods Human type Ⅱ alveolar epithelial cell line A549 was induced by 5 ng/mL TGF-β1 in vitro for 72 hours and divided into control, TGF-β1, and 10-8 mol/L Ac-SDKP intervention groups. Morphological changes of epithelial-mesenchymal transition in A549 cells were observed by phase-contrast microscopy. Immunocytochemical methods were used to determine the locations of cytokeratin 8 (CK8) and vimentin. The expressions of E-cadherin (E-cad), CK8, vimentin, α-smooth muscle actin (α-SMA), HSP27, and type Ⅰ and type Ⅲ collagen were detected by Western blot analysis.

    Results The A549 cells formed spindle-like after exposed to TGF-β1. Coincident with these morphological changes, the expression levels of CK8 and E-cad decreased to 40% and 50% as compared to the control group, while those of vimentin and α-SMA increased to 1.9 and 1.8 folds respectively (P<0.05). This process was accompanied by increases in levels of HSP27, type Ⅰ collagen, and type Ⅲ collagen by 1.9, 2.2, and 2.5 folds (P<0.05). After Ac-SDKP in tervention, the expressions of CK8 and E-cad increased by 2.6 folds and 2.0 folds respectively as compared to the TGF-β1 group; while the expressions of vimentin, α-SMA, HSP27, type Ⅰcollagen, and Ⅲ were decreased to 68%, 66%, 74%, 66%, and 44% comparing with the TGF-β1 group (P<0.05).

    Conclusion Ac-SDKP could inhibit the transition of cultured human type Ⅱ alveolar epithelial cells to myofibroblasts and attenuate collagen synthesis through modulating the expression of HSP27.

     

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