唾液体外诱导促进兔脂肪干细胞上皮分化  

Epithelial differentiation of adipose stem cells induced by saliva in vitro

作　　者：李晓文 许颖捷 贾梦莹 史玮 贾馨雨 龚忠诚[1,2] Li Xiaowen;Xu Yingjie;Jia Mengying;Shi Wei;Jia Xinyu;Gong Zhongcheng(Oncological Department of Oral&Maxillofacial Surgery,First Affiliated Hospital(Affiliated Stomatological Hospital)of Xinjiang Medical University,Urumqi 830054,Xinjiang Uygur Autonomous Region,China;Xinjiang Uyghur Autonomous Region Institute of Stomatology,Urumqi 830054,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]新疆医科大学第一附属医院(附属口腔医院)颌面肿瘤外科,新疆维吾尔自治区乌鲁木齐市830054 [2]新疆维吾尔自治区口腔医学研究所,新疆维吾尔自治区乌鲁木齐市830054

出　　处：《中国组织工程研究》2022年第24期3865-3869,共5页Chinese Journal of Tissue Engineering Research

基　　金：自治区研究生科研创新项目(XJ2020G193),项目负责人:李晓文。

摘　　要：背景:颊脂垫常用于修复口腔颌面部缺损,其能使口腔黏膜快速上皮分化。由于缺损创面暴露于唾液中,因而认为唾液可能促进颊脂垫中的脂肪干细胞上皮化。目的:探讨唾液体外诱导促进兔脂肪干细胞上皮分化的可行性。方法:取两三个月龄的新西兰大白兔脂肪组织,分离、培养脂肪干细胞并传代。将第3代脂肪干细胞分为3组,即阴性对照组、表皮生长因子组与唾液组,后两组采用10μg/L表皮生长因子、15%唾液来诱导脂肪干细胞,倒置显微镜观察细胞形态变化,诱导7,14 d采用免疫荧光、RT-PCR检测细胞角蛋白19的表达。结果与结论:①唾液诱导后脂肪干细胞形态由长梭形逐渐变为短梭形,甚至不规则的扁圆形;②诱导7,14 d,表皮生长因子组、唾液组细胞角蛋白19呈阳性表达,均高于阴性对照组,且诱导14 d细胞角蛋白19的表达量明显高于诱导7 d;③诱导7,14 d,表皮生长因子组、唾液组细胞角蛋白19的mRNA表达量均高于阴性对照组(P<0.01),且诱导14 d的表达量要显著高于诱导7 d(P<0.01);④结果表明,唾液在一定程度上能促进脂肪干细胞向上皮分化。BACKGROUND:Buccal fat pads are often used to repair oral and maxillofacial defects,which can rapidly differentiate the oral mucosa from the epithelium.Because the defect wound is exposed to saliva,it is believed that saliva may promote the epithelialization of adipose stem cells in the buccal fat pad.OBJECTIVE:To investigate the feasibility of promoting epithelial differentiation of rabbit adipose stem cells by induction of saliva in vitro.METHODS:Fat harvested from 2-3 months New Zealand white rabbit was isolated,cultured,generated adipose stem cells for passage.Passage 3 adipose stem cells were divided into three groups:negative control group,epidermal growth factor group and saliva group.Adipose stem cells were induced by 10μg/L epidermal growth factor and 15%saliva in the epidermal growth factor group and saliva group.Cell morphology changes were observed under inverted microscope.The expression of cytokeratin 19 was detected by immunofluorescence and RT-PCR after 7 and 14 days of induction.RESULTS AND CONCLUSION:(1)After saliva induction,the morphology of adipose stem cells was gradually changed from long spindle shape to short spindle shape or even irregular oblate shape.(2)At 7 and 14 days,cytokeratin 19 was positively expressed in the epidermal growth factor and saliva groups,which was higher than that in the negative control group.The expression level of cytokeratin 19 in the 14-day induction group was significantly higher than that in the 7-day induction group.(3)The expression of cytokeratin 19 mRNA in epidermal growth factor and saliva groups was higher than that in negative control group(P<0.01).The expression level at 14 days was significantly higher than that at 7 days(P<0.01).(4)Results suggest that saliva can promote the epithelial differentiation of adipose stem cells to some extent.

关 键 词：干细胞 脂肪干细胞 表皮生长因子 唾液 上皮 分化 细胞角蛋白19 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]