KLF2对山羊肌内前体脂肪细胞分化的影响

doi:10.11843/j.issn.0366-6964.2020.01.008

Abstract

Abstract: The aim of this study was to clone the sequence of zinc finger domain of goat KLF2, to clarify the tissue and cell expression profiles of KLF2, and finally to elucidate the effect of interfering KLF2 gene on the goat intramuscular preadipocytes differentiation and the possible ways of action. In this study, 5 healthy, 1-year-old Jianzhou Big Ear goats were selected as experiment animals. By using RT-PCR, cell culture, real-time quantitative PCR (qPCR), RNA interference and other methods, the sequence of the zinc finger domain of goat KLF2 gene was cloned, the tissue and cell temporal expression of KLF2 gene was detected and futher the effect of KLF2 gene on goat intramuscular preadipocytes differentiation was elucidated. The results showed that the length of goat KLF2 zinc finger domain was 454 bp (KU041748.1), and the domain had 100% homology with sheep, cattle and mice. KLF2 was widely expressed in goat tissues with the higher expression level in the lung and abdominal fat (P<0.05). The expression of KLF2 decreased first, then increased, and decreased again during the intramuscular preadipocytes differentiation, the expression level reached to the lowest after differentiation 12 h, which was significantly lower than that in undifferentiated preadipocytes (P<0.05). Oil red O staining result showed that interfering KLF2 significantly promoted the accumulation of lipid droplets in adipocytes; and the expression of PPARγ and C/EBPβ were extremely significantly up-regulated by 48.5% (P<0.01) and significantly up-regulated by 43.6%(P<0.05); At the same time, the expression level of KLF1, KLF13, KLF14, KLF15 and KLF16 were extremely markedly increased (P<0.01), the expression level of KLF4 was significantly decreased (P<0.05), and the expression level of KLF3, KLF6, KLF8, KLF9 and KLF11 were extremely significantly decreased (P<0.01). The zinc finger domain of goat KLF2 was highly conserved with other species, and KLF2 was highly expressed in lung and abdominal fat of goat(P<0.05). Meanwhile, KLF2 was a negative regulator for goat intramuscular preadipocytes differentiation, and might play an important role through targeting PPARγ and C/EBPβ genes, what's more it could hierarchically regulated other KLFs members.

Key words: goat, KLF2, RNA interference, intramuscular preadipocytes differentiation, expression mode

CLC Number:

HE Changsheng, WANG Yong, XU Qing, BAI Wenlin, WANG Jianglin, ZHU Jiangjiang, LIN Yaqiu. The Effect of KLF2 on the Differentiation of Goat Intramuscular Preadipocyte[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(1): 64-73.

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The Effect of KLF2 on the Differentiation of Goat Intramuscular Preadipocyte

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