Kruppel样因子15对和盈黑鸡前体脂肪细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2022.07.009

摘要/Abstract

摘要： 旨在明确和盈黑鸡Kruppel样因子15(KLF15)的组织、细胞表达模式，阐明干扰KLF15对和盈黑鸡前体脂肪细胞增殖分化的影响。本研究采用RT-qPCR技术检测KLF15在各组织中表达分布、在腹脂组织的发育性表达规律以及在成脂诱导分化不同时期细胞中的表达水平；干扰KLF15后，采用CCK-8和EdU法检测细胞增殖，利用油红O染色、甘油三酯含量分析以及脂肪细胞分化标志基因(PPARγ、C/EBPa和FAS)的检测，分别从细胞形态学和分子生物学等角度明确干扰KLF15对和盈黑鸡前体脂肪细胞成脂分化的影响。研究结果表明，KLF15在和盈黑鸡各组织中存在广泛表达，在肝、腿肌和腹脂组织中表达量较高；KLF15在公、母鸡腹脂中分别呈“L”和“U”型表达趋势，表达高峰均出现在1日龄；KLF15 mRNA在细胞分化前期缓慢升高，表达高峰出现在细胞分化后期；细胞增殖检测发现，CCK-8和EdU结果趋势一致，干扰KLF15极显著抑制了前体脂肪细胞的增殖(P＜0.01)；明显减少了前体脂肪细胞的脂滴积聚，极显著降低了甘油三酯含量(P＜0.01)，且脂肪细胞分化标志基因PPARγ和C/EBPa mRNA水平极显著下调(P＜0.01)，FAS mRNA水平显著下调(P＜0.05)。本研究揭示了和盈黑鸡KLF15的表达模式，验证了干扰KLF15能够抑制前体脂肪细胞的增殖分化，推测KLF15是前体脂肪细胞的负调控因子，可能是通过调控PPARγ、C/EBPa和FAS等分化关键基因的表达来实现的。

关键词: 和盈黑鸡, KLF15, 前体脂肪细胞, 干扰, 细胞增殖与分化

Abstract: The purpose of this study was to clarify the tissue and cell expression pattern of KLF15 in Heying Black chickens and to elucidate the effects of interfering KLF15 on the proliferation and differentiation of preadipocytes of Heying Black chickens. RT-qPCR was used to detect the expression of KLF15 in various tissues, the developmental expression pattern in abdominal fat tissues and the expression level in cells at different stages of adipogenic induction differentiation. After interfering with KLF15, cell proliferation was detected by CCK-8 and EdU, triglyceride content was analyzed, Oil red O staining was carried out, and marker genes for adipocyte differen-tiation were detected (PPARγ, C/EBPa and FAS). The effects of interfering with KLF15 on adipogenic differentiation of Heying Black chicken preadipocytes were investigated from morphological and molecular biological perspectives. The results showed that KLF15 was widely expressed in all detected tissues of Heying Black chickens, with higher expression levels in liver, leg muscle and abdominal fat tissues. KLF15 showed "L" and "U" shaped expression trend in abdominal fat of males and females, respectively, and the expression peak appeared at one day of age. KLF15 mRNA increased slowly in the early stage of cell differentiation, and its expression peak occurred in the late stage of cell differentiation. Cell proliferation detection showed that the results of CCK-8 and EdU exhibited the same trend, and interfering with KLF15 highly significantly inhibited the proliferation of preadipocytes (P<0.01), obviously reduced the accumulation of lipid droplets in preadipocytes, extremely significantly reduced triglyceride content (P<0.01). In addition, the mRNA levels of PPARγ and C/EBPa were extremely significantly down-regulated (P<0.01), and FAS mRNA levels were significantly down-regulated (P<0.05). This study revealed the expression pattern of KLF15 in Heying Black chickens and verified that interfering with KLF15 could inhibit the proliferation and differentiation of preadipocytes. It is speculated that KLF15 is a negative regulator of preadipocytes, it possibly regulate the differentiation of preadipocytes through regulating the expression of PPARγ, C/EBPa and FAS genes.

Key words: Heying Black chicken, KLF15, preadipocytes, interference, cell proliferation and differentiation

中图分类号:

S831.2

陈兰, 张涛, 丁浩, 谢恺舟, 张跟喜, 王金玉. Kruppel样因子15对和盈黑鸡前体脂肪细胞增殖分化的影响[J]. 畜牧兽医学报, 2022, 53(7): 2118-2129.

CHEN Lan, ZHANG Tao, DING Hao, XIE Kaizhou, ZHANG Genxi, WANG Jinyu. Effects of Kruppel-like Factor 15 Gene on Proliferation and Differentiation of Preadipocytes of Heying Black Chickens[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2118-2129.

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