牵张力作用下瘢痕疙瘩间充质干细胞对成纤维细胞增殖及胶原合成的影响  

Effect of Cultured Supernatant from Mechanical-stretch-treated Keloid-derived Mesenchymal Stem Cells on Keloid Fibroblasts Proliferation and Collagen Synthesis

作　　者：周大昂 宋海峰[1] 刘涛[1] 曹天宇[1] 张衍国[1] 

机构地区：[1]第四军医大学唐都医院皮肤科,陕西西安710038

出　　处：《中国皮肤性病学杂志》2017年第7期714-717,722,共5页The Chinese Journal of Dermatovenereology

基　　金：陕西省自然科学基金(2015SF164)

摘　　要：目的研究牵张力作用下瘢痕疙瘩间充质干细胞(keloid-derived mesenchymal stem cells,KD-MSCs)培养上清对瘢痕疙瘩成纤维细胞(keloid fibroblasts,KFs)增殖和胶原合成的影响,探讨牵张力及KD-MSCs在瘢痕疙瘩形成过程中的作用。方法培养原代KD-MSCs和KFs,于72h收集牵张力作用下和未施加牵张力的KD-MSCs培养上清,并分别孵育KFs 24h,48h,72h,96h,倒置相差显微镜观察细胞的形态学变化,CCK-8法检测KFs的增殖活力,Real-time PCR检测KFs中Ⅰ型和Ⅲ型胶原和CTGF mRNA转录情况,羟脯氨酸消化法检测胶原水平的变化。结果培养上清刺激后,两组间KFs形态未见明显差异;CCK-8结果提示牵张力组KFs增殖明显强于无牵张力组(P<0.05);Real-time PCR结果示牵张力组KFsⅠ型胶原的mRNA表达明显高于无牵张力组(P<0.01),牵张力组KFsⅢ型胶原的mRNA表达低于无牵张力组(P<0.05),牵张力组KFs CTGF的mRNA表达明显低于无牵张力组(P<0.01);羟脯氨酸检测提示牵张力组KFs羟脯氨酸含量明显高于无牵张力组(P<0.05)。结论牵张力作用可明显增强KD-MSCs培养上清对KFs增殖和胶原合成的促进作用,提示牵张力作用下KD-MSCs可发生旁分泌促进成纤维细胞增生和分泌,从而参与瘢痕疙瘩的形成,其具体机制仍需深入研究。Objective To investigate the effect of cultured supernatant from mechanical-stretch-treated keloid-derived mesenchymal stem cells（KD-MSCs）on keloid fibroblasts（KFs）proliferation and collagen synthesis and to explore the role of mechanical stretch and KD-MSCs in the formation of keloid. Methods KD-MSCs were treated with or without mechanical stretch, and the supernatant was collected on 72h. KFs were cultured with or with out KD-MSCs supernatant. After 24h, 48h, 72h and 96h, the cell morphology was observed under inverted phase contrast microscope, the proliferation activity was detected with CCK-8 kit, the mRNA expression levels of collagen typeⅠ（ColⅠ）, collagen type Ⅲ（ColⅢ）and connective tissue growth factor（CTGF）were detected by Real-time PCR, and the level of collagen changes were detected by hydroxyproline assay kit（hyp）. Results After being treated with KD-MSCs supernatant, no significant difference were found between the two groups; CCK-8 showed that the proliferation level of KFs in tension group was higher than that in no tension group（P〈0.05）; Real-time PCR showed a higher mRNA expression of collagenⅠ（P〈0.01）, and lower mRNA expressions of collagen Ⅲ（P〈0.05）and CTGF（P〈0.01）; hyp showed that hydroxyproline content was higher in tension group than that in no tension group（P〈0.05）. Conclusion Treatment of mechanical stretch enhanced the capacity of KD-MSCs supernatant to promote KFs proliferation and collagen secretion, suggesting that mechanical-stretch-treated KD-MSCs promote the proliferation and secretion of KFs by paracrine, thus contribute to the keloid formation. However, the underlying mechanism needs to be further explored.

关 键 词：瘢痕疙瘩 间充质干细胞 牵张力 成纤维细胞 

分 类 号：R739.5[医药卫生—肿瘤] R329.2[医药卫生—临床医学]