Effects of lncFAM200B on the lipid deposition in intramuscular preadipocytes of yak.
Effects of lncFAM200B on the lipid deposition in intramuscular preadipocytes of yak.
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DOI：
10.3864/j.issn.0578-1752.2022.13.014
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发表时间：
2022-01-01
影响因子：
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通讯作者：
Zhong, J. C.
中科院分区：
文献类型：
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作者：
Ran, Hong-biao;Zhao, Li-ling;Zhong, J. C.
目的：本研究旨在分析lncFAM 200 B对牦牛肌内前脂肪细胞脂质沉积的影响，为进一步的机制研究奠定基础。方法：取牦牛背最长肌组织，分离肌内前脂肪细胞。利用腺病毒介导的过表达技术实现lncFAM 200 B的过表达，并利用siRNA干扰技术分析lncFAM 200 B的功能。采用实时荧光定量PCR（RT-qPCR）检测脂肪分化标志物PPARgamma、C/EBPalpha和AP 2的mRNA表达水平以及lncFAM 200 B潜在靶基因SIRT 1和PTEN的mRNA表达水平。油红O染色、三酰甘油（TAG）测定和CCK-8测定法检测细胞内脂滴沉积和前脂肪细胞增殖。测试结果：过表达lncFAM 200 B不仅显著增加了脂肪分化相关基因（C/EBPalpha和AP 2，P < 0.05）的表达水平，而且在诱导分化过程中增加了细胞内大脂滴的沉积。相反，lncFAM 200 B干扰降低了PPARgamma、C/EBPalpha和AP 2的表达（P < 0.05），以及脂滴沉积。另外，lncFAM 200 B过表达4d后，三酰甘油含量显著高于对照组（P < 0.05），而siRNA干扰6d后，三酰甘油含量显著低于对照组（P < 0.05）。在lncFAM 200 B过表达过程中，SIRT 1表达水平随时间呈先降低后升高的趋势，而PTEN表达水平随时间呈先降低后升高的趋势，而在lncFAM 200 B干扰过程中，两种基因表达水平随时间呈先降低后升高的趋势。CCK-8实验结果显示，无论是过表达组还是干扰组，72 h后细胞增殖活性均有显著性差异。结论：lncFAM 200 B可以通过影响脂肪分化标志物C/EBPalpha和AP 2的表达来调控牦牛脂肪分化，进而影响三酰甘油含量和脂滴沉积，但具体机制有待进一步研究。
Objective: The aim of this study was to analyze the effects of lncFAM200B in the lipid deposition in intramuscular preadipocytes of yak, which laid a foundation for further mechanism research. Method: The longissimus dorsi muscle tissue of yak was collected and used to separate intramuscular preadipocytes. The adenovirus mediated overexpression technology was used to realize the overexpression of lncFAM200B, and the siRNA interference technology was used to analyze the function of lncFAM200B. The mRNA expression level of fat differentiation marker (PPARgamma, C/EBPalpha and AP2) and the potential target (SIRT1 and PTEN) genes of lncFAM200B were detected via real-time fluorescent quantitative PCR (RT-qPCR). Oil red O staining, triacylglycerol (TAG) determination and CCK-8 determination methods were used to detect intracellular lipid droplet deposition and preadipocyte proliferation. Result: The overexpression of lncFAM200B not only significantly increased the fat differentiation genes (C/EBPalpha and AP2, P < 0.05) expression level, but also increased the lipid droplets deposition with large lipid droplets in the cells during induced differentiation. Conversely, lncFAM200B interference reduced the expression of PPARgamma, C/EBPalpha and AP2 (P < 0.05), and lipid droplet deposition. Furthermore, the triacylglycerol content after lncFAM200B overexpression 4 days was significantly higher than that of the control group (P < 0.05), but lower after siRNA interference 6 days. Moreover, the SIRT1 levels increased with time was first decreased and then increased trend, and the PTEN had the opposite trend during lncFAM200B overexpression, and the opposite results of the two genes obtained during lncFAM200B interference. In addition, the results of CCK-8 experiment showed that there was a significant difference in cell proliferation activity both in overexpression or interference group after 72 hours. Conclusion: lncFAM200B could regulate yak adipose differentiation by influencing the expression of fat differentiation marker (C/EBPalpha and AP2), and further affect the triacylglycerol content and the lipid droplet deposition, but the detail mechanism need to be further research efforts.