| 蒋 硕,陈思远,孟轩羽,刘迪晖,梁小弟.诱导小鼠白色脂肪来源的SVF分化为米色脂肪细胞的方法初探[J].,2023,(10):1822-1828 |
| 诱导小鼠白色脂肪来源的SVF分化为米色脂肪细胞的方法初探 |
| The Method of Inducing White Adipose-derived SVF to Differentiate into Beige Adipocytes |
| 投稿时间：2023-01-09 修订日期：2023-02-05 |
| DOI：10.13241/j.cnki.pmb.2023.10.004 |
| 中文关键词: SVF细胞 米色脂肪细胞 UCP1 PRDM16 鸡尾酒诱导法 |
| 英文关键词: SVF cells Beige fat cells UCP1 PRDM16 Cocktail inducement |
| 基金项目:国家自然科学基金地区项目（81760162）；2017年度高校科研计划面上项目（XJEDU2017M016）；新疆维吾尔自治区研究生科研创新项目（XJ2022G184） |
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| 中文摘要: |
| 摘要 目的：探究将小鼠白色脂肪来源的SVF细胞诱导为米色脂肪细胞的方法，为米色脂肪的研究提供细胞模型。方法：分离培养小鼠脂肪组织来源的SVF细胞，观察其细胞形态及生长特性，分别用鸡尾酒法和米色脂肪诱导法处理细胞，在诱导的第0、2、4、6、8、10天收取细胞样品进行Western blot和RT-qPCR实验，油红O染色观察不同时间点培养皿中的脂质生成情况，并对针对脂滴大小定量。结果：实验分离的SVF细胞随着接种时间的延长逐渐呈长梭形，在+接种120 h后有呈螺旋式生长的趋势。Western blot和RT-qPCR结果显示两种诱导方法皆使过氧化物酶体增殖物受体γ（Peroxisomeproliferator activated receptor γ，PPARγ）和CCAAT-增强子结合蛋白（CCAAT/enhancer binding proteins α，C/EBPα）基因表达升高（P<0.05），油红O染色结果显示，在两种方法的诱导下，脂质生成水平无差异，但与鸡尾酒法相比，米色脂肪诱导法的脂肪细胞脂滴面积明显减少（P<0.05），并且小体积脂滴所占百分比较高。Western blot和RT-qPCR结果显示，与鸡尾酒法相比，米色脂肪诱导法使解偶联蛋白1（Uncoupling protein 1，UCP1）和含PR结构域蛋白16（PRD1-BF1-RIZ1 homologous domain containing 16，PRDM16）基因表达升高（P<0.05）。结论：与鸡尾酒法相比，该方法能够成功诱导SVF细胞向米色脂肪细胞分化，脂滴具备米色脂肪特征，并表达米色脂肪标记基因。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the method of inducing mice SVF cells derived from white fat into beige fat cells, and to provide a cell model for the study of beige fat. Methods: SVF cells derived from mouse adipose tissue were isolated and cultured, and their cell morphology and growth characteristics were observed. The cells were treated with cocktail method and beige fat induction method, respectively, and cell samples were collected at the 0, 2, 4, 6, 8 and 10 days of induction for Western blot and RT-qPCR. Lipid production in the petri dishes at different time points was observed by oil red O staining, and lipid drop size was quantified. Results: The isolated SVF cells showed a long spindle shape gradually, and a spiral growth trend after 120 h inoculation. Western blot and RT-qPCR results indicated that Peroxisome proliferator activated receptor γ (PPARγ) could be induced by both methods. The gene expressions of PPARγ and CCAAT/enhancer binding proteins α (C/ EBP-α) were increased (P<0.05). Oil red O staining showed no difference in lipid production levels under the induction of the two methods, but compared with that in the cocktail method, Fat droplet area of fat cells induced by beige fat decreased significantly(P<0.05), and the percentage of small fat droplet was higher. Western blot and RT-qPCR results showed that compared with that by the cocktail method, the beige fat induction method enabled the uncoupling protein 1 (UCP1) and an increased gene expression of PRD1-BF1-RIZ1 homologous domain containing 16 (PRDM16) (P<0.05). Conclusion: Compared with the cocktail method, this method can induce SVF cells to differentiate into beige adipocytes successfully. The adipocytes have the characteristics of beige adipocytes and express the beige adipocyte marker genes. |
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