人牙龈间充质干细胞条件培养液促进大鼠皮肤创面愈合的实验研究  

The experimental study of human gingival mesenchymal stem cell conditioned medium promoting skin wound healing in rats

作　　者：张洁 杨华军 赵伟 蒋巧平 李敏 胡胜杰 Zhang Jie

机构地区：[1]杭州市儿童医院,310005 [2]杭州市拱墅区朝晖街道社区卫生服务中心,310014

出　　处：《浙江临床医学》2023年第9期1287-1289,1293,共4页Zhejiang Clinical Medical Journal

基　　金：杭州市医药卫生科技项目(A20200798)。

摘　　要：目的探究人牙龈间充质干细胞条件培养液(GMSCs-CM)对大鼠皮肤创面愈合的影响及作用机制。方法将20只大鼠随机分为GMSCs-CM组和PBS组。建立皮肤切割伤模型后于大鼠皮肤创伤周围4个部位共注射GMSCs-CM或PBS液200μL(15μg/mL),1次/d,连续3 d。分别在注射第3、7、14天时观察创面愈合情况,计算创面愈合率。通过HE染色检测病理学改变,免疫组化检测CD31的表达,Western blot检测PCNA、TGF-b1和VEGF蛋白的表达。结果第7、14天时GMSCs-CM组的创面愈合率明显高于PBS组。病理学显示GMSCs-CM组成纤维细胞、胶原纤维及血管增多,炎性细胞浸润增加。免疫组化显示GMSCs-CM组创面CD31染色阳性的细胞数量增多。Western blot显示第3、7、14天创面PCNA、TGF-b1和VEGF蛋白表达量均显著高于PBS组(P<0.05)。结论注射GMSCs-CM可通过上调VEGF和TGF-b1的表达,促进成纤维细胞增殖、毛细血管和肉芽组织的形成,从而加快皮肤创面的愈合。Objective To investigate the effect and mechanism of human gingival mesenchymal stem cell conditioned medium(GMSCs-CM)on skin wound healing in rats.Methods Twenty rats were randomly divided into two groups:GMSCs-CM group and PBS control group.After establishing the skin incision model,GMSCs-CM or PBS 200μL(15μg/mL)was injected into 4 sites around the rats skin wound,once a day for 3 consecutive days.The wound healing was observed at 3,7 and 14 days after injection,and the wound healing rate was calculated.Pathological changes were detected by HE staining,the expressions of CD31 were detected by immunohistochemistry,and the expressions of PCNA,TGF-b1 and VEGF proteins were detected by Western blot.Results The wound healing rate of the GMSCs-CM group was significantly higher than that of the PBS control group at 7,14 days.Compared with the control group,the pathological results showed that the fibrocytes,collagen fibers and blood vessels of GMSCs-CM were increased,and the inflammatory cells were increased.Immunohistochemistry showed an increased number of cells with positive wound CD31 staining in the GMSCs-CM group.Western blot showed that the protein expressions of PCNA,TGF-b1 and VEGF in GMSCs-CM group were significantly higher than those in PBS group at 3,7 and 14 days(P<0.05).Conclusion Injection of GMSCs-CM can promote the proliferation of fibroblasts and increase the formation of capillaries and granulation tissue by up-regulating the expression of VEGF and TGF-b1,thus accelerating the healing of skin wounds.

关 键 词：牙龈间充质干细胞 条件培养基 创面愈合 血管内皮生长因子、转化生长因子β1 

分 类 号：R285.5[医药卫生—中药学]