摘要
目的:探讨人脂肪干细胞(hADSCs)在皮肤创伤愈合中的作用机制。方法:构建裸鼠皮肤创伤模型,随机分成hADSCs组和对照组,分别于创缘周围注射hADSCs细胞和磷酸盐缓冲液,观察并记录创面大小,取术后创缘组织,行HE染色、实时定量RT-PCR、Western blot法检测Notch通路相关因子表达;体外实验转染Jagged1过表达质粒至hADSCs细胞,Notch1过表达质粒至人角质形成细胞(HaCaT),构建hADSCs和HaCaT细胞共培养体系,使用CCK8、Transwell及划痕实验检测HaCaT细胞增殖和迁移能力,实时定量RT-PCR、Western blot法检测Notch信号通路相关因子表达。结果:术后3 d两组裸鼠皮肤创面均开始结痂,术后21 d hADSCs组的愈合率明显高于对照组(P<0.05),术后4、7 d hADSCs组的上皮再生量均明显高于对照组(P<0.05),hADSCs组Notch信号通路相关因子表达量显著升高(P<0.05)。转染过表达质粒并共培养后,HaCaT细胞的增殖、迁移能力显著增强,Notch通路被激活(P<0.05)。结论:hADSCs通过Jagged1/Notch信号通路促进皮肤创伤愈合。
To explore the role of human adipose-derived stem cells(hADSCs)in skin wound healing and to investigate the potential mechanism.Methods:Nude mice skin trauma models were constructed and subsequently divided into hADSCs group,and control group.The trauma size was observed and recorded at the indicated time.HE staining was performed to observe the regeneration of the epithelium.The real time quantitative PCR and Western blot were also used for the detection of Notch downstream pathway expression level.The hADSCs and HaCaT cells were co-cultured and were transfected with Jagged1 and Notch1 overexpression plasmid respectively,and Notch downstream pathway expression levels were detected by real time quantitative RT-PCR and Western blot.The proliferation and migration of HaCaT cells were detected by CCK8,transwell and scratch test.Results:All groups began to scab after 3 days of the molding.The healing rates was significantly higher than control group on day 21,respectively(P<0.05).The regenerated epidermis of hADSCs group was significantly higher than control group on day 4 and day 7 after the molding(P<0.05).And Notch downstream pathway expression level was significantly higher in the hADSCs group(P<0.05).After transfection with overexpression plasmid and co-culture,the proliferation and migration ability of HaCaT cells were significantly enhanced,Notch pathways were activated.Conclusions:hADSCs can promote skin wound healing process through Jagged1/Notch signaling pathway.
作者
王义
郑旸
董孟杰
江丰
王超
宋晓萌
WANG Yi;ZHENG Yang;DONG Mengjie;JIANG Feng;WANG Chao;SONG Xiaomeng(Department of Oral and Maxillofacial Surgery, The Affiliated Stomatological Hospital of Nanjing Medical University, Jiangsu Province Key Laboratory of Oral Diseases, Jiangsu Province Engineering Research Center of Stomatological Translational Medicine, Nanjing 210029, China)
出处
《口腔生物医学》
2022年第1期9-14,共6页
Oral Biomedicine
基金
国家自然科学基金(81772877)
江苏高校优势学科建设工程资助项目(2018-87)
江苏省医学创新团队资助项目(CXTDA-2017036)。
