1α, 25-dihydroxy Vitamin D3 containing fractions of Catharanthus roseus leaf aqueous extract inhibit preadipocyte differentiation and induce lipolysis in 3T3-L1 cells

Abstract

BackgroundTo investigate the potential of Catharanthus roseus leaf aqueous crude extract (CRACE) as a regulator of adipocyte development and function.Methods3T3-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.ResultsCRACE 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.Conclusions1α, 25-dihydroxy Vitamin D3 containing CRACE can be developed into an effective anti-obesity formulation that decreases adipogenesis and increases lipid catabolism.

In vivo and in vitro effects of chronical exposure to nonylphenol on lipid metabolism

“…The exposure dose increased, the red color became deeper, and the fat cells were more densely distributed; WB results showed the protein expressions of CEBPα, FAS, SREBP1, and PPARγ increased in adipocytes. 3T3-L1 pre-adipocyte in vitro experiments are often used to evaluate the interference of environmental chemicals on adipogenesis, and provide a basis for environmental substances that can affect lipid metabolism in the body [5,10,24]. Studies have shown that [11,17]: 4-NP increased PPARγ activity at 1 μM, and significantly promoted triglyceride accumulation in 3T3-L1 adipocytes at 10 μM.…”

Mechanism of action of Indonesian medicinal plants in inhibiting 3T3-L1 adipocyte differentiation: A review

“…In the early stages of differentiation, there was a decrease in the level of stimulatory Lipin1 and an increase in the level of the inhibitory Kruppel-like factor (KLF7). Meanwhile, at the final stage of differentiation, there was a decrease in levels of C/EBPα, Lipin1, and other similar positive effectors such as fatty acid synthase (FAS) and sterol regulatory element binding protein 1c (SREBP1c) in the extract-treated cells (Borah et al, 2019). O n l i n e F i r s t…”

“…This plant is most commonly found in Java, Sulawesi, Kalimantan, and Sumatra islands. It has various benefits, especially as a medicinal plant, where various properties were identified (Borah et al, 2019;Nejat et al, 2015). Catharanthus roseus is known to have anti-diabetic, anti-microbial, anti-mutagenic, anti-mitotic, antioxidant, and anticancer effects (Nejat et al, 2015).…”

“…It contains many alkaloids and phenolics, which are the main components of this plant, but others are also found, such as alkaloids, flavonoid glycosides, polyphenols, anthocyanins, steroids, and iridoid glucosides (Nejat et al, 2015). Based on research from Borah et al (2019) the extract from the leaves of the plant can inhibit the adipogenesis process in 3T3-L1 cells.…”

“…Catharanthus roseus leaf extract was able to inhibit the expression of PPARγ1 and PPARγ2 messenger-RNA and protein. This was evidenced by the decreased levels of adipocyte markers affected by PPARγ, such as perilipin 1 (PLN1), fatty acid binding protein 4 (FABP4), lipoprotein lipase (LPL), adipose triglyceride O n l i n e F i r s t lipase (ATGL), glucose transporter 4 (GLUT4), and adiponectin (Borah et al, 2019). Various transcription factors, both stimulatory and inhibitory for the adipogenesis process, were influenced by leaf extracts.…”