| As a type of growth factor, VEGF have been proved to promote hair growth in mouse. VEGFR-2 can regulate the fuction of VEGF as the primary receptor. It have been proved that VEGF can promote the growth of human dermal papilla cells through the activation of Erkl/2 signal pathway via VEGFR-2.In this research, the cashmere production improved (10048) and unimproved (10056) transgenic cashmere goats were used as experimantal group, while the somatic cell cloning goat (NT) from the same cell line was used as control group, the expression of VEGF and VEGFR-2 in the skin tissues and VEGF related signal pathway genes was detected by real time-PCR and Western Blot.The mRNA expression of VEGF and related signal pathway genes in goat skin tissues of NT,10056 and 10048 was detected by real time-PCR. Results showed that the expression of VEGF in skin tissues was no significant differences between 10056 and NT, while the expression of VEGFR-2 in 10056 was significantly higher than that of NT (P<0.05). The expression of VEGF in skin tissue of 10048 was significantly higher than that of NT (P<0.05), and the expression of VEGFR-2 in 10048 was significantly higher than that of NT (P<0.01). VEGF related signal pathway genes including PI3K, Akt, RelA, β-catenin and HIF1-α were detected. Results showed that the expression of β-catenin and PI3K in skin tissue of 10048 was no significant difference than that of NT, the expression of HIF1-α was significantly higher than that of NT (P<0.05). The expression of β-catenin in skin tissue of 10056 was significantly higher than that of NT (P<0.05), the expression of HIF1-α and PI3K in skin tissue of 10056 was significantly lower than that of NT (P<0.05).The protein expression of VEGF, VEGFR-2 and related signal pathway genes were detected by Western Blot. Results showed that, the expression of VEGF, VEGFR-2, Erk1/2, P-Erk, Akt P-Akt, P38, P-P38 in skin tissue of 10056 was 1.4,0.9,1.54,1.14,0.96,1.92,1.3,3 times of NT respectively. The expression of VEGF, VEGFR-2, Erk1/2, P-Erk, Akt P-Akt, P38, P-P38 in skin tissue of 10048 was 1.1,3.1,1.74,1.1,0.9, 1.24,1.19,2.26 times of NT respectively.The results suggested that, the expression of VEGF and VEGFR-2 was up-regulated in skin tissue of cashmere production improved transgenic goat (10048) compared to somatic cell clone goat (NT), while the expression of Erk1/2 and P-Erk 1/2 was also up-regulated to activate the Erk1/2 signal pathway, and caused the up-regulation of P-P38 and P-Akt phosphorylation.Meanwhile, SHF-DPCs of inner mongolia arbas cashmere goat in anagen were used to dectect the effects of VEGF overexpression and interference on expression of VEGF, VEGFR-2 and related signal pathway genes.The mRNA expression of VEGF and VEGFR-2 in VEGF overexpressed and down-regulated anagen SHF-DPCs was detected by real time-PCR. The results showed that, mRNA expression of VEGF in SHF-DPCs electroporated by VEGF 164 interference 1 plasmid (1-1) was significantly lower than that of SHF-DPCs electroporated by negtive control plasmid (NC). The mRNA expression of VEGF in SHF-DPCs electroporated by VEGF 164 interference 2 plasmid (1-2) was lower than that of NC. The mRNA expression of VEGF in SHF-DPCs electroporated by VEGF 164 overexpressing plasmid (O-V) was significantly higher than that of NC; the mRNA expression of VEGFR-2 was higher than that of NC. Protein expression of VEGF and VEGFR-2 in VEGF overexpressed and down-regulated SHF-DPCs was detected by Western Blot. The results showed that, the expression of VEGF in SHF-DPCs electroporated by 1-1 was 0.6 times of that of NC, the expression of VEGFR-2 was 0.69 times of that of NC. The expression of VEGF in SHF-DPCs electroporated by 1-2 was 0.86 times of that of NC, the expression of VEGFR-2 was 0.92 times of that of NC.The expression of VEGF in SHF-DPCs electroporated by O-V was 1.14 times of that of NC,the expression of VEGFR-2 was 1.4 times of that of NC.Protein expression of VEGF, VEGFR-2 related signal pathway genes (Erkl/2, P38, Akt) in VEGF overexpressed and down-regulated SHF-DPCs was detected by Western Blot. The results showed that, the expression of Erkl/2, P38, Akt in SHF-DPCs electroporated by VEGF 164 interference 1 plasmid (1-1) were 0.82,1.01,0.59 times of that of NC. The expression of Erkl/2, P38, Akt in SHF-DPCs electroporated by VEGF 164 interference 2 plasmid (1-2) were 0.92,0.89,0.32 times of that of NC. The expression of Erkl/2, P38, Akt in SHF-DPCs electroporated by VEGF 164 overexpressing plasmid (O-V) were 0.84,1.44,0.69 times of that of NC.The results indicated that the expression of VEGFR-2 was positive correlation with the expression of VEGF when VEGF was overexpressed and down-regulated in SHF-DPCs. These results provide experimental evidences to understand the related signal pathways of VEGF during the hair follicle development of Inner Mongolia cashmere goat. |
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