摘要
目的研究海地瓜硫酸软骨素(Acaudina Molpadioideschondroitin sulfate,AM-CHS)对3T3-L1前脂肪细胞增殖和分化的影响,并探讨其作用机制。方法采用传统的鸡尾酒诱导剂诱导分化3T3-L1前脂肪细胞,以MTT法检测AM-CHS对3T3-L1前脂肪细胞及不同分化阶段3T3-L1细胞增殖活性的影响;分别采用油红O染色和甘油三酯(triglycerides,TG)含量测定法评价其对3T3-L1前脂肪细胞分化的影响。采用RT-PCR法检测脂肪细胞中过氧化物酶体增殖体激活受体γ(peroxisome proliferators-activated receptors gamma,PPARγ)、CCAAT增强子结合蛋白α(CCAAT/enhancer bind-ing protein alpha,C/EBPα)、固醇调节元件结合蛋白-1c(ste-rol regulatory element binding protein-1c,SREBP-1c)等分化相关基因的mRNA表达水平。结果 AM-CHS能明显抑制3T3-L1前脂肪细胞和成熟脂肪细胞的增殖,抑制3T3-L1前脂肪细胞的分化过程,以对分化早期的抑制作用最强。RT-PCR结果表明,AM-CHS能明显降低脂肪细胞PPARγ、C/EBPα和SREBP-1c mRNA的表达。结论海地瓜AM-CHS能明显抑制3T3-L1前脂肪细胞的增殖和分化,其作用机制与下调分化相关基因PPARγ、C/EBPα和SREBP-1c的表达有关。
Aim To observe the effect of AM-CHS on the proliferation and differentiation of 3T3-L1 preadipocytes, and to investigate its possible mechanism. Methods After using differentiation cocktail to differentiate 3T3-L1 preadipocytes, the effects of AM-CHS on the proliferation activities of 3T3-L1 cells at different time were determined by MTT assay. Besides, the effect of AM-CHS on the differentiation of 3T3-L1 preadipocytes was carried out by oil red O staining and TG content determination. Expression levels of PPARγ, C/EBPγ and SREBP-1c mRNA were measured by reverse transcription PCR（RT-PCR）. Results AM-CHS inhibited the proliferation of 3T3-L1 preadipoeytes and mature adipocytes, and inhitited the differentiation of 3T3-L1 preadipocytes, in which the stron- gest inhibition appeared in the early state. RT-PCR resuits showed that AM-CHS down-regulated the expression of PPARγ, C/EBPα and SREBP-1 c mRNA, which presented time-dose independence. Conclusion AM-CHS can markedly suppress the proliferation and differentiation of 3T3-L1 preadipocytes, and the underlying mechanisms may be related to down-regulating the expression of PPARγ, C/EBPα and SREBP-1 c mRNA.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2013年第5期708-714,共7页
Chinese Pharmacological Bulletin
基金
"十二五"国家科技支撑计划(No 2012BAD33B07)
海洋公益性行业科研专项(No 201105029)
长江学者和创新团队发展计划资助
