畜牧兽医学报2018，Vol.49Issue(6)：1124-1133,10.DOI:10.11843/j.issn.0366-6964.2018.06.004
VEGF对体外培养绒山羊次级毛囊外根鞘细胞的影响
Effect of VEGF on Secondary Hair Follicle Outer Root Sheath Cells of Cashmere Goat in vitro
摘要
Abstract
This study was conducted to investigate isolation and cultivation of the hair follicle outer root sheath cells(HFORSCs)in vitro,and to estimate the effects of the vascular endothelial growth factor(VEGF)on the proliferation,differentiation and mRNA expression of proliferating cell nuclear antigen(PCNA),keratin 10(K10)and fibroblast growth factor 5(FGF 5)genes in the sHFORSCs of Shaanbei White cashmere goat.Skin tissues of goats were collected,and sHFOR-SCs were obtained through the methods of digestion and then cultured under the condition of 37 ℃,5％CO2and 100％relative humidity ;The 5thgeneration of cultured sHFORSCs were selected and treated with different concentration VEGF for 24 and 48h,respectively.The cell vi-ability was detected by Methyl thiazolyl tetrazolium(MTT),and the cell proliferation was detected by EdU DNA markers ;The mRNA expression levels of PCNA,K10 and FGF5 in HFORSCs were detected by the real-time fluorescence quantitative PCR.Results showed that :1)sHFORSCs were successfully isolated and cultured in vitro,and cultured cells were positive in the immunohistochemistry test of CK 17 and CK 15,which were main biomarkers of outer root sheath cells.2)VEGF could promote the proliferation of sHFORSCs.Treated with same concen-tration of VEGF,the cell viability of 48 h group was significantly higher than that of 24 h group(P＜0.01).Treated with different concentrations of VEGF for 48 h,the cell viability of 100 ng·mL－1 group was significantly higher than that of control group,1 ng·mL －1group and 10 ng·mL －1 group(P＜0.01),and also significantly higher than that of 50 ng·mL －1group(P＜0.05).The cell viability of 50ng·mL－1group was significantly higher than that of control group(P＜0.05);The proportion of the cell proliferation of 100 and 50 ng· mL －1groups were all significantly higher than that of control group,1 ng·mL－1group and 10 ng·mL－1group(P＜0.01).The proportion of the cell proliferation of 100 ng·mL－1group was significantly higher than that of 50ng·mL－1group(P＜0.05).3)The expressions of mRNA of PCNA,K10 and FGF5 were all detected in HFOR-SCs.Treated with VEGF for 48 h,the mRNA expression level of PCNA in 100 ng·mL－1group was the highest,and was significantly higher than that in control group(P＜0.01)and 10 ng·mL －1 group(P＜ 0.05);The mRNA expression of K10 in control group was the highest,and was significantly higher compared with that in 10 and 100ng·mL －1groups(P＜0.01);The mRNA expression of FGF 5 in control group was the highest,and was significantly higher than that in 100ng·mL－1group(P＜0.01).In this study,sHFORSCs were successfully isolated from hair follicle and were successfully cultured in vitro ;The proliferation of HFORSCs could be promoted and the cell differentiation could be inhibite by VEGF.It inferred that VEGF might play the role of grow th promotion of the hair follicle by inhibiting the mRNA expression of FGF 5 in sHFORSCs.关键词
绒山羊/血管内皮生长因子/次级毛囊外根鞘细胞/增殖/成纤维细胞生长因子5/mRNA表达Key words
cashmere goats/vascular endothelial growth factor/secondary hair follicle outer root sheath cells/proliferation/fibroblast growth factor 5/mRNA expression分类
农业科技引用本文复制引用
张婧婧,王德光,周小兵,高晔,何晓琳,陈玉林,张恩平..VEGF对体外培养绒山羊次级毛囊外根鞘细胞的影响[J].畜牧兽医学报,2018,49(6):1124-1133,10.基金项目
国家重点研发计划(2016YFD0500508) （2016YFD0500508）
陕西省重点研发计划(201TSCXL-NY-04-02) （201TSCXL-NY-04-02）
陕西省农业科技创新转化项目(NYKJ-2016-04) （NYKJ-2016-04）
