摘要
背景:脂肪细胞分化是一个复杂的生物学过程,涉及前脂肪细胞向成熟脂肪细胞的转化,这一过程在肥胖和相关代谢性疾病的发生发展中发挥关键作用。近年来,C1q/肿瘤坏死因子相关蛋白家族作为脂肪因子新成员,已成为代谢调控领域的研究热点。目的:探究C1q肿瘤坏死因子相关蛋白4对3T3-L1前脂肪细胞分化的作用及机制。方法:采用CCK-8法测定不同质量浓度C1q肿瘤坏死因子相关蛋白4对3T3-L1前脂肪细胞活性的影响,选择安全浓度进行干预。将3T3-L1前脂肪细胞分为对照组、诱导剂组、 C1q肿瘤坏死因子相关蛋白4组,首先将3T3-L1前脂肪细胞进行接触抑制2 d,随后加入成脂诱导剂和C1q肿瘤坏死因子相关蛋白4处理。分化第10天,采用油红O染色观察脂滴形成程度,采用RT-qPCR和Western blot检测CCAAT/增强子结合蛋白α、过氧化物酶体增殖物激活受体γ、脂肪酸结合蛋白4基因的mRNA和蛋白表达水平,采用甘油三酯和总胆固醇检测试剂盒测定细胞内甘油三酯和总胆固醇水平。结果与结论:C1q肿瘤坏死因子相关蛋白4对3T3-L1前脂肪细胞无毒性作用的最高安全质量浓度为1 000 ng/mL。与对照组相比,诱导剂组细胞内脂滴形成增加,甘油三酯和总胆固醇水平升高(P<0.05),CCAAT/增强子结合蛋白α、过氧化物酶体增殖物激活受体γ、脂肪酸结合蛋白4的mRNA及蛋白表达水平显著上调(P<0.05)。与诱导剂组相比,C1q肿瘤坏死因子相关蛋白4组细胞内脂滴形成减少,甘油三酯和总胆固醇水平降低(P<0.05);CCAAT/增强子结合蛋白α、过氧化物酶体增殖物激活受体γ、脂肪酸结合蛋白4的mRNA及蛋白表达水平显著下调(P<0.05)。结果表明,C1q肿瘤坏死因子相关蛋白4能抑制前脂肪细胞分化为成熟脂肪细胞,作用机制可能与下调CCAAT/增强子结合蛋白α、过氧化物酶体增殖物激活受体γ、脂肪酸结合蛋白4的表达有关。
BACKGROUND:Adipocyte differentiation is a complex biological process involving the transformation of preadipocytes into mature adipocytes.This process plays a key role in the development and progression of obesity and related metabolic diseases.In recent years,the complement C1q/tumor necrosis factorrelated protein family,as a new member of the adipokine family,has become a research hotspot in the field of metabolic regulation.OBJECTIVE:To investigate the effect of complement 1q/tumor necrosis factor-related protein 4 on the differentiation of 3T3-L1 preadipocytes and its underlying mechanism.METHODS:The CCK-8 assay was used to determine the effects of different concentrations of complement 1q/tumor necrosis factor-related protein 4 on the viability of 3T3-L1 preadipocytes,and a safe concentration was selected for intervention.3T3-L1 preadipocytes were divided into a control group,an inducer group,and a complement 1q/tumor necrosis factor-related protein 4 group.3T3-L1 preadipocytes were first contact-inhibited for 2 days,followed by adipogenic inducer and complement 1q/tumor necrosis factor-related protein 4 treatment.On day 10 of differentiation,lipid droplet formation was observed using Oil Red O staining.The mRNA and protein expression levels of CCAAT/enhancer binding proteinα,peroxisome proliferator-activated receptorγ,and fatty acid binding protein 4 were determined by RT-qPCR and western blot assay.Intracellular triglyceride and total cholesterol levels were measured using triglyceride and total cholesterol detection kits.RESULTS AND CONCLUSION:The safe maximum concentration of complement 1q/tumor necrosis factor-related protein 4 without toxicity to 3T3-L1 preadipocytes was 1000 ng/mL.Compared with the control group,the inducer group showed increased intracellular lipid droplet formation,elevated triglyceride and total cholesterol levels(P<0.05),and significantly upregulated mRNA and protein expression levels of CCAAT/enhancer binding proteinα,peroxisome proliferator-activated receptorγ,and fatty acid binding protein 4(P<0.05).Compared with the inducer group,the complement 1q/tumor necrosis factor-related protein 4 group showed reduced intracellular lipid droplet formation,lower triglyceride and total cholesterol levels(P<0.05).The mRNA and protein expression levels of CCAAT/enhancer binding proteinα,peroxisome proliferator-activated receptorγ,and fatty acid binding protein 4 were significantly downregulated(P<0.05).These results suggest that complement 1q/tumor necrosis factor-related protein 4 inhibits the differentiation of preadipocytes into mature adipocytes,and its mechanism of action may be related to downregulating the expression of CCAAT/enhancer binding proteinα,peroxisome proliferator-activated receptorγ,and fatty acid binding protein 4.
作者
买热艳木·肉孜
汪红平
张翠平
夏娟
申甜
雷涛
鲁郡
高洁
Maireyanmu·Rozi;Wang Hongping;Zhang Cuiping;Xia Juan;Shen Tian;Lei Tao;Lu Jun;Gao Jie(Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China)
出处
《中国组织工程研究》
北大核心
2026年第19期4867-4872,共6页
Chinese Journal of Tissue Engineering Research
基金
普陀区卫生系统科技创新项目(ptkwws202417),项目负责人:高洁
普陀区卫生系统科技创新项目(ptkwws202003),项目负责人:雷涛
普陀区卫生系统科技创新项目(ptkwws202302),项目负责人:鲁郡
普陀区临床特色专科(2024tszk02),项目负责人:鲁郡
普陀区临床特色专科(2020tszk01),项目负责人:雷涛
上海市卫生健康委员会科研项目(202240309),项目负责人:鲁郡。
