目的 探究脑源性神经营养因子(BDNF)对幼儿脂肪细胞增殖和分化的影响,为儿童肥胖症的治疗提供借鉴。方法 采集正常和肥胖儿童脂肪,real-time qPCR和Western blotting分别检测脂肪组织中BDNF、补体C1q肿瘤坏死因子相关蛋白13(CTRP13)和IL-6 mRNA以及蛋白的表达;分离培养原代前体脂肪细胞,采用不同浓度的BDNF和BDNF siRNA分别处理细胞,MTT和油红O染色分别检测细胞增殖和分化,甘油三酯( TG) 测定试剂盒检测细胞成脂情况,Western blotting 技术检测PPARγ、aP2、CTRP13和IL-6的表达。结果 与正常组相比,肥胖组脂肪组织中IL-6的表达较高,而CTRP13和BDNF的表达较低;BDNF对脂肪细胞的增殖没有影响;BDNF促进CTRP13的表达,抑制脂肪细胞分化和IL-6的表达,而BDNF siRNA则作用相反。结论 肥胖儿童脂肪组织中IL-6表达增高,CTRP13和BDNF表达降低,BDNF可抑制幼儿原代脂肪细胞的分化。
Abstract
Objective To explore the effects of brain derived neurotrophic factor (BDNF) on proliferation and differentiation of child preadipocytes,and provide guidance for the treatment of childhood obesity. Methods Adipose tissue were collected from normal and obese children,real-time qPCR and western blotting were used respectively for measuring the expression of mRNA and protein of BDNF,C1q/TNF-related protein-13(CTRP13) and IL-6;Preadipocytes were isolated from adipose tissue and cultured with different concentrations of BDNF and BDNF siRNA respectively,the level of preadipocytes' proliferation was determined by MTT and the differentiation by oilred O,triglyceride (TG) assay kits was used for detecting the intracellular lipogenesis,the levels of PPARγ,aP2,CTRP13 and IL-6 were determined by western blotting. Results The obesity children showed higher expression of IL-6 in adipose tissue compared to control group,while lower expression of CTRP13 and BDNF in obesity children.BDNF had no effects for proliferation of children preadipocyte; BDNF promoted the expression of CTRP13 in children preadipocytes,while inhabited IL-6 and differentiation of preadipocytes.Moreover,BDNF siRNA showed the opposite effect with BDNF. Conclusion IL-6 was overexpression in obesity children,but BDNF and CTRP13 are lower in obesity children compared to the normal,BDNF can inhibit the differentiation of preadipocytes.
关键词
儿童肥胖症 /
脑源性神经营养因子 /
脂肪细胞分化 /
补体C1q肿瘤坏死因子相关蛋白13 /
白细胞介素6
Key words
obesity of children /
brain derived neurotrophic factor /
adipogenic differentiation /
C1q/TNF-related protein-13 /
interleukin-6
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