VEGF对体外培养绒山羊次级毛囊外根鞘细胞的影响

doi:10.11843/j.issn.0366-6964.2018.06.004

Abstract

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 sHFORSCs were obtained through the methods of digestion and then cultured under the condition of 37℃, 5% CO₂ and 100% relative humidity;The 5^th generation of cultured sHFORSCs were selected and treated with different concentration VEGF for 24 and 48 h, respectively. The cell viability was detected by Methyl thiazolyl tetrazolium (MTT), and the cell proliferation was detected by EdU DNA markers;The mRNA expression levels of PCNA, K 10 and FGF 5 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 CK17 and CK15, which were main biomarkers of outer root sheath cells. 2) VEGF could promote the proliferation of sHFORSCs. Treated with same concentration 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^-1 group and 10 ng·mL^-1 group (P<0.01), and also significantly higher than that of 50 ng·mL^-1 group (P<0.05). The cell viability of 50 ng·mL^-1 group was significantly higher than that of control group (P<0.05); The proportion of the cell proliferation of 100 and 50 ng·mL^-1 groups were all significantly higher than that of control group, 1 ng·mL^-1 group and 10 ng·mL^-1 group (P<0.01). The proportion of the cell proliferation of 100 ng·mL^-1 group was significantly higher than that of 50 ng·mL^-1 group (P<0.05). 3) The expressions of mRNA of PCNA, K10 and FGF 5 were all detected in HFORSCs. Treated with VEGF for 48 h, the mRNA expression level of PCNA in 100 ng·mL^-1 group 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 100 ng·mL^-1 groups (P<0.01); The mRNA expression of FGF 5 in control group was the highest, and was significantly higher than that in 100 ng·mL^-1 group (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 inhibited by VEGF. It inferred that VEGF might play the role of growth promotion of the hair follicle by inhibiting the mRNA expression of FGF 5 in sHFORSCs.

Key words: cashmere goats, vascular endothelial growth factor, secondary hair follicle outer root sheath cells, proliferation, fibroblast growth factor 5, mRNA expression

CLC Number:

S827.2

ZHANG Jing-jing, WANG De-guang, ZHOU Xiao-bing, GAO Ye, HE Xiao-lin, CHEN Yu-lin, ZHANG En-ping. Effect of VEGF on Secondary Hair Follicle Outer Root Sheath Cells of Cashmere Goat in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(6): 1124-1133.

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Effect of VEGF on Secondary Hair Follicle Outer Root Sheath Cells of Cashmere Goat in vitro

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