KLF11抑制山羊肌内前体脂肪细胞分化

doi:10.11843/j.issn.0366-6964.2020.05.009

摘要/Abstract

摘要： 旨在获得山羊Krüppel样因子11（Krüppel-like factor 11，KLF11）基因序列，明确其组织及细胞表达模式，阐明KLF11对山羊肌内前体脂肪细胞分化的影响及可能作用途径。本研究利用实时荧光定量PCR（quantitative real-time PCR，qPCR）技术检测KLF11在山羊各组织及成脂诱导分化不同阶段细胞中的表达水平，利用油红O染色和qPCR等方法从形态学及分子生物学等角度确定干扰KLF11后对山羊肌内脂肪细胞分化的影响及可能的作用途径。结果表明，KLF11核苷酸序列总长为1 752 bp，CDS区为1 518 bp，编码505个氨基酸残基。KLF11基因在山羊各个组织中都有广泛表达，并且在肝、背最长肌及腹部脂肪中存在较高水平表达；KLF11在成脂诱导48 h的山羊肌内脂肪细胞中表达水平极显著高于诱导分化前（P<0.01）。筛选到干扰效率可达到63.58%的KLF11-siRNA1有效干扰序列，转染山羊肌内前体脂肪细胞后发现可明显促进山羊肌内脂细胞的脂滴积聚，且脂肪细胞分化标志基因PPARγ和C/EBPβ mRNA水平出现极显著上调（P<0.01），Pref-1 mRNA水平出现极显著下调（P<0.01）；干扰KLF11基因后，KLFs有些成员（KLF1、KLF2、KLF4、KLF5、KLF8、KLF10、KLF14、KLF16）表达水平发生极显著下降（P<0.01），有些（KLF15）则出现相反的表达模式。KLF11基因可能通过调控PPARγ、C/EBPβ和Pref-1的表达抑制山羊肌内前体脂肪细胞分化，并且这种作用可能是通过协同KLF2、KLF4、KLF10，拮抗KLF7和KLF15来进行的。

关键词: 山羊, KLF11基因, 肌内前体脂肪细胞, 干扰, 细胞分化

Abstract: This study was performed to obtain the sequence of Krüppel-like factor 11 (KLF1) gene, clarify its expression pattern in tissues and cells of goats, and further elucidate its effects on the differentiation of goat intramuscular preadipocytes with its possible action pathway. The expression of KLF11 in goat different tissues and cells at different adipogenic differentiation stages was detected by real-time fluorescence quantitative PCR (qPCR). From both morphology and molecular biology, Oil red O staining and qPCR were used to reveal the effect of interfering KLF11 on the differentiation of intramuscular adipocytes of goats with its possible action pathway. The results showed that KLF11 gene was 1 752 bp in length, CDS region was 1 518 bp in length, which encoded 505 amino acids. KLF11 gene was widely expressed in the various tissues of goats. The expression level of KLF11 was higher in liver, longissimus dorsi and abdominal fat. Compared with the preadipocytes, the expression of KLF11 gene extremely significantly increased at 48 h of differentiation(P<0.01).After obtaining the effective interference sequence of KLF11-siRNA1, the expression of KLF11 was significantly decreased by KLF11-siRNA1 to 63.58%, obviously promoted accumulation of lipid droplets were odserved in the intramuscular adipocytes of goats. After interfering KLF11, the mRNA levels of PPARγ and C/EBPβ, the adipocyte differentiation marker genes were extremely significantly up-regulated (P<0.01), while the expression of Pref-1 was extremely significantly down-regulated (P<0.01). Meanwhile, the expression levels of some members of KLFs family (KLF1,KLF2,KLF4,KLF5,KLF8,KLF10,KLF14,KLF16) significantly declined and others increased after interfering KLF11 gene. KLF11 gene could inhibit the differentiation of goat intramuscular preadipocytes through regulating the expression of PPARγ, C/EBPβ and Pref-1, and this effect might be achieved by synergizing with KLF2, KLF4, KLF10 and antagonizing with KLF7, KLF15.

Key words: goat, KLF11 gene, intramuscular preadipocytes, RNAi, cell differentiation

中图分类号:

S827.2

王江林, 王永, 池永东, 朱江江, 林亚秋. KLF11抑制山羊肌内前体脂肪细胞分化[J]. 畜牧兽医学报, 2020, 51(5): 976-986.

WANG Jianglin, WANG Yong, CHI Yongdong, ZHU Jiangjiang, LIN Yaqiu. KLF11 Inhibits the Differentiation of Goat Intramuscular Preadipocyte[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 976-986.

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