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

作　　者：邓海静[1] 李世峰[1] 孙月[2] 张丽娟[1] 薛新新 杜世璞 徐洪[1] 杨方[1] 

机构地区：[1]河北联合大学实验研究中心,老年医学国际科技合作基地,河北063000 [2]沧州市青县人民医院,河北062650

出　　处：《环境与职业医学》2015年第2期113-117,共5页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(编号:81072254;81302395);河北省高等学校科学技术研究重点项目(编号:ZD20131035);唐山市科技计划项目任务书(编号:13130299z)

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/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 N-acetyl-seryl-aspartyl-lysyl-proline （Ac-SDKP） could inhibit epithelial- mesenchymal transition of human type Ⅱ alveolar epithelial cells through suppressing the expression of heat shock protein27 （HSP27） and the deposition of type Ⅰ and type Ⅱ 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 （CKS） and vimentin. The expressions of E-cadherin （E-cad）, CKS, vimentin, a-smooth muscle actin （α-SMA）, HSP27, and type I and type m 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 a-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 intervention, 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 Ⅰ collagen, and Ⅲ were decreased to 68%, 66%, 74%, 66%, and 44% comparing with tbe 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.

关 键 词：N-乙酰基-丝氨酰-天门冬氨酰-赖氨酰-脯氨酸 上皮-间质转化 肺泡Ⅱ型上皮细胞 肌成纤维细胞 热休克蛋白27 

分 类 号：R114[医药卫生—卫生毒理学]