胰岛素样生长因子1促进毛囊间充质干细胞增殖的机制研究  

Mechanism study on insulin-like growth factor 1 in the promotion of hair follicle mesenchymal stem cells prolif⁃eration

作　　者：夏颖 丁娟[1] 张建青[1] 陈宏[1] XIA Ying;DING Juan;ZHANG Jianqing(Department of Plastic Surgery,Shaoxing People′s Hospital,Shao xing 312000,China.)

机构地区：[1]绍兴市人民医院整形美容科,浙江绍兴312000

出　　处：《全科医学临床与教育》2021年第10期874-878,F0002,共6页Clinical Education of General Practice

基　　金：浙江省基础公益研究计划(LGD19H110002)。

摘　　要：目的探讨胰岛素样生长因子1(IGF-1)对毛囊间充质干细胞(HF-MSCs)的增殖作用及其可能机制。方法采用酶消化法从大鼠触须中获取HF-MSCs,用流式细胞术对其进行了表面标记的鉴定,采用多向诱导分化实验鉴定其多潜能性。用不同浓度的IGF-1以及p38抑制剂(SB203580)、ERK1/2抑制剂(U0126)、JNK抑制剂(SP600125)处理细胞,MTT实验检测HF-MSCs增殖的改变。采用Western blot检测HF-MSCs中p38、ERK1/2和JNK信号通路的激活情况。结果分离的HF-MSCs表面表达间充质干细胞的标记物CD44、CD73、CD90、CD105,不表达内皮细胞标志物CD31。HF-MSCs具有向骨细胞和脂肪细胞分化的能力。MTT实验结果显示,干预24 h时,25 nM IGF-1组和50 nM IGF-1组细胞OD值较对照组增加(t分别=5.96、8.46,P均<0.05);干预48 h、72 h和96 h时,10 nM IGF-1组、25 nM IGF-1组和50 nM IGF-1组细胞OD值较对照组增加(t分别=5.69、9.23、11.10、7.88、11.50、14.00、11.10、13.60、16.90,P均<0.05),且呈剂量依赖性(F分别=5.43、6.03、7.94,P均<0.05)。Western blot结果显示,与对照组比较,10 nM IGF-1组p-p38磷酸化水平增加;25 nM IGF-1组和50 nM IGF-1组的p38和ERK1/2磷酸化水平提高,而JNK磷酸化水平无改变。进一步MTT实验结果显示,干预48 h、72 h和96 h时,与IGF-1组比较,IGF-1+SB203580和IGF-1+U0126组的细胞OD值下降(t分别=5.28、6.17、7.91、9.04、6.85、7.22,P均<0.05),而IGF-1+SP600125组细胞OD值无变化(t分别=0.93、1.34、1.05,P均>0.05)。结论IGF-1可能通过磷酸化IGF-1R,进一步激活p38和ERK1/2通路促进HF-MSCs增殖。Objective To explore the effect of insulin-like growth factor 1(IGF-1)on the proliferation of hair follicle mesenchymal stem cells(HF-MSCs)and its potential mechanisms.Methods HF-MSCs was obtained from rat tentacles by enzymatic digestion,and flow cytometry was used to identify their surface markers,multidirectional induced differentia-tion experiments were used to identify their pluripotency.Cells were treated by various concentration of IGF-1,as well as p38 inhibitor(SB203580),ERK1/2 inhibitor(U0126)and JNK inhibitor(SP600125).The proliferation of HF-MSCs was examined by MTT assay.Western blot was used to detect the activation of p38,ERK1/2 and JNK signaling pathways in HF-MSCs.Results The isolated HF-MSCs exhibited surface markers of mesenchymal stem cells including CD44,CD73,CD90 and D105,and not exhibited endothelial marker CD31.Subsequent experiments showed the differentiation po-tentials towards adipocytes and osteoblasts of HF-MSCs.MTT assay showed that at 24h of intervention,the optical density(OD)values of cells in the 25 nM IGF-1 group and 50 nM IGF-1 group were significantly increased compared with the control group(t=5.96,8.46,P<0.05),at 48h,72h and 96h of intervention,the OD values of cells in the 10 nM IGF-1 group,25 nM IGF-1 group and 50 nM IGF-1 group were significantly increased compared with the con-trol group(t=5.69,9.23,11.10,7.88,11.50,14.00,11.10,13.60,16.90,P<0.05),in a dose-dependent manner(F=5.43,6.03,7.94,P<0.05).Western blot showed that compared with the control group,the expression of phosphorylated p38 was increased in the 10 nMIGF-1 group,the levels of phosphorylated p38 and phosphorylated ERK1/2,but not phosphorylated JNK was significantly in-creased in the 25 nM IGF-1 group and 50 nM IGF-1 group.In addition,MTT assay found that the OD values of cells was decreased in IGF-1+SB203580 and IGF-1+U0126 groups when compared with IGF-1 group(t=5.28,6.17,7.91,9.04,6.85,7.22,P<0.05),whereas,there was no statistical difference in the change the OD values of cells in IGF-1+SP600125 group(t=0

关 键 词：毛囊间充质干细胞 胰岛素样生长因子1 增殖 

分 类 号：R622[医药卫生—整形外科]