山羊KLF16对肌内前体脂肪细胞分化的影响

Effect of KLF16on the differentiation of goat intramuscular preadipocytes

为分析山羊KLF16对肌内前体脂肪细胞分化的调控作用,利用RT-PCR克隆得到山羊KLF16基因序列,利用生物信息学分析山羊KLF16基因序列特征,通过过表达、油红O染色和qPCR等方法从形态学及分子水平分析过表达山羊KLF16后肌内脂肪细胞脂滴积聚及分化标志基因表达水平的变化。结果表明:1)克隆得到包含CDS区的山羊KLF16基因序列986bp,编码251个氨基酸,具有典型锌指结构;2)KLF16在山羊各组织广泛表达,其中腹部皮下脂肪组织表达量极显著高于其他组织(P<0.01);3)过表达山羊KLF16与对照组相比脂肪细胞脂滴积聚减少,分化标志基因PPARγ和PPARα极显著下调(P<0.01),C/EBPβ显著下调(P<0.05);4)过表达山羊KLF16后KLFs部分成员发生显著的上调或下调,相关性分析结合转录因子结合位点预测结果推测KLF16可能通过拮抗KLF8来发挥其对分化的调控作用。综上,山羊KLF16可能通过拮抗KLF8抑制分化标志基因PPARγ、PPARα和C/EBPβ的表达,从而抑制肌内前体脂肪细胞分化,为揭示山羊KLF16调控脂肪细胞分化提供重要的数据支持。

In order to analyze the regulation of goat KLF16 on the differentiation of intramuscular precursor adipocytes,the goat KLF16 gene sequence was cloned by RT-PCR,and the characteristics of goat KLF16 gene sequence were analyzed by bioinformatics.Through overexpression,oil red O staining,qPCR and bioinformatics analysis,the changes in lipid droplet accumulation and differentiation marker gene expression levels in intramuscular adipocytes after overexpression of goat KLF16 were clarified from the morphological and molecular levels.he results showed that:1)The cloned goat KLF16 gene sequence containing the CDS region was 986 bp encoding 251 amino acids with a typical zinc finger structure;2)KLF16 was ubiquitous expressed in goat tissues,and the expression of abdominal subcutaneous fat was significantly higher(P<0.01);3)Compared with the control group,the overexpression of goat KLF16 decreased the accumulation of lipid droplets in adipocytes.The marker genes PPARγ and PPARα were extremely significantly down-regulated(P<0.01),and C/EBPβ was significantly down-regulated(P<0.05);4)Some members of KLFs were significantly up-regulated or down-regulated after overexpression of goat KLF16.Correlation analysis combined with the prediction of transcription factor binding sites speculated that KLF16 may regulate goat intramuscular adipocyte differentiation by antagonizing KLF8.In summary,goat KLF16 may inhibit the differentiation marker gene PPARγ,PPARαand C/EBPβ by antagonizing KLF8.This study provided important data support for revealing the KLF16 regulating adipocyte differentiation in goats.