BACKGROUND To investigate the potential of Catharanthus roseus leaf aqueous crude extract (CRACE) as a regulator of adipocyte development and function. METHODS 3T3-L1 adipogenesis model was used to investigate the effect of CRACE on adipogenesis. 3T3-L1 preadipocytes (for adipogenic differentiation) and mature 3T3-L1 adipocytes (for adipocyte function) were treated with non-toxic doses of CRACE. The outcomes were corroborated by intracellular lipid accumulation, expression of pro-and anti-adipogenic effector molecules. To investigate CRACE mediated lipolysis, cAMP accumulation, glycerol release and phosphorylation of key effector molecules were tested in treated mature adipocytes. Finally, the extract was fractionated to identify the active molecule/s in the extract. RESULTS CRACE significantly reduced adipocyte differentiation by modulating PPARγ expression. At early stage CRACE directly targeted Lipin1 expression and consequently impacted KLF7, subsequently expression of GATA2, CEBPα, SREBP1c were targeted, with PPARγ expression, particularly curtailed. While CRACE significantly reduced several lipogenic genes like FAS and GPD1 in mature adipocytes, concomitantly, it greatly increased lipolysis resulting in decreased lipid accumulation in mature adipocytes. The increase in lipolysis was due to decreased Akt activation, increased cAMP level, and PKA activity. The fractionation of CRACE allowed identification of two fractions with potent anti-adipogenic activity. Both the fractions contained 1α, 25-dihydroxy Vitamin D3 as major component. CONCLUSIONS 1α, 25-dihydroxy Vitamin D3 containing CRACE can be developed into an effective anti-obesity formulation that decreases adipogenesis and increases lipid catabolism.

中文翻译：

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长春花叶水提取物中含有1α，25-二羟基维生素D3的组分抑制3T3-L1细胞中的前脂肪细胞分化并诱导脂解。

背景技术研究长春花叶片含水粗提取物（CRACE）作为脂肪细胞发育和功能调节剂的潜力。方法采用3T3-L1脂肪形成模型研究CRACE对脂肪形成的影响。用无毒剂量的CRACE处理3T3-L1前脂肪细胞（用于脂肪分化）和成熟的3T3-L1脂肪细胞（用于脂肪细胞功能）。细胞内脂质蓄积，促脂肪和促脂肪形成的效应分子的表达证实了这一结果。为了研究CRACE介导的脂解作用，在处理过的成熟脂肪细胞中测试了cAMP的积累，甘油的释放和关键效应分子的磷酸化。最后，将提取物分级分离以鉴定提取物中的活性分子。结果CRACE通过调节PPARγ表达显着降低了脂肪细胞的分化。在早期阶段，CRACE直接靶向Lipin1表达，进而影响KLF7，随后靶向GATA2，CEBPα，SREBP1c的表达，尤其是减少了PPARγ的表达。虽然CRACE显着减少了成熟脂肪细胞中的一些脂肪生成基因，例如FAS和GPD1，但同时，它大大增加了脂解作用，从而导致了成熟脂肪细胞中脂质的积累减少。脂肪分解的增加是由于Akt激活降低，cAMP水平升高和PKA活性降低所致。CRACE的分馏可鉴定出具有有效抗脂肪形成活性的两个馏分。两种级分均包含1α，25-二羟基维生素D 3作为主要成分。结论1α，