Yonsei Med J.  2018 Jan;59(1):85-91. 10.3349/ymj.2018.59.1.85.

Adipogenic and Lipolytic Effects of Ascorbic Acid in Ovariectomized Rats

• Kim B
• Choi KM
• Yim HS
• Park HT
• Yim JH
• Lee MG

• KMID: 2418853
• DOI: http://doi.org/10.3349/ymj.2018.59.1.85

Abstract

PURPOSE
Ascorbic acid has been reported to have an adipogenic effect on 3T3-L1 preadipocytes, while evidence also suggests that ascorbic acid reduces body weight in humans. In this study, we tested the effects of ascorbic acid on adipogenesis and the balance of lipid accumulation in ovariectomized rats, in addition to long-term culture of differentiated 3T3-L1 adipocytes.
MATERIALS AND METHODS
Murine 3T3-L1 fibroblasts and ovariectomized rats were treated with ascorbic acid at various time points. In vitro adipogenesis was analyzed by Oil Red O staining, and in vivo body fat was measured by a body composition analyzer using nuclear magnetic resonance.
RESULTS
When ascorbic acid was applied during an early time point in 3T3-L1 preadipocyte differentiation and after bilateral ovariectomy (OVX) in rats, adipogenesis and fat mass gain significantly increased, respectively. However, lipid accumulation in well-differentiated 3T3-L1 adipocytes showed a significant reduction when ascorbic acid was applied after differentiation (10 days after induction). Also, oral ascorbic acid administration 4 weeks after OVX in rats significantly reduced both body weight and subcutaneous fat layer. In comparison to the results of ascorbic acid, which is a well-known cofactor for an enzyme of collagen synthesis, and the antioxidant ramalin, a potent antioxidant but not a cofactor, showed only a lipolytic effect in well-differentiated 3T3-L1 adipocytes, not an adipogenic effect.
CONCLUSION
Taking these results into account, we concluded that ascorbic acid has both an adipogenic effect as a cofactor of an enzymatic process and a lipolytic effect as an antioxidant.

Keyword

Ascorbic acid; obesity; ovariectomy; adipogenesis; lipolysis; 3T3-L1 cell

MeSH Terms

3T3-L1 Cells
Adipocytes/drug effects/metabolism
Adipogenesis/*drug effects
Animals
Antioxidants/pharmacology
Ascorbic Acid/*pharmacology
Body Composition/drug effects
Body Weight/drug effects
Cell Differentiation/drug effects
Female
Fibroblasts/drug effects/metabolism
Lipolysis/*drug effects
Mice
*Ovariectomy
Rats, Sprague-Dawley
Antioxidants
Ascorbic Acid

Figure

• 

  Fig. 1 Long-term effect of ASC on adipogenesis of 3T3-L1 mouse preadipocytes. (A) Graphical demonstration of long-term adipogenesis with or without ASC by ORO staining. Adipogenic induction on Day 0 and ASC treatment during the early period (Day -2, Day 0, and Day 2). (B) Effect of ASC on long-term adipogenesis depending on the treatment period. ORO staining was measured on Day 14. ASC treatment during the early period, the late period (Day 8, Day 10, and Day 12), or both (n=9). (C) Western blotting analysis of ASC treated adipocytes on Day 14 against collagens, α1(I) and α1(VI), and adipogenic markers, PPARγ and CEBPα. ASC treatment during the early period or both early and late periods. (D) Immunocytochemical features of ASC treated adipocytes against type I collagen at Day 14. ASC treatment during the early period, or both early and late periods. Arrow indicates the area of unstained cells. *p<0.05, **p<0.01. ASC, ascorbic acid; ORO, Oil Red O; OD, optical density.

• 

  Fig. 2 Inhibitory effect of the antioxidant ramalin on adipogenesis of 3T3-L1 mouse preadipocytes. Adipogenesis was induced with or without ASC during the early period (Day -2, Day 0, and Day 2). Ramalin treatment during the early or late period (Day 8, Day 10, and Day 12). (A) ORO staining picture of ramalin treated adipocytes on Day 14. (B) Graphical demonstration of adipogenesis in ramalin-treated adipocytes with ORO staining on Day 14 (n=9). *p<0.05, **p<0.01. ASC, ascorbic acid; ORO, Oil Red O; OD, optical density.

• 

  Fig. 3 Body composition analysis of ovariectomized rats with or without ASC injection. ASC injection during the early period (Week 1 and 2), the late period (Week 5 and 6), or both after OVX (n=8). (A) Body weight difference 6 weeks after OVX. (B) Body fat difference 6 weeks after OVX. *p<0.05, **p<0.01. ASC, ascorbic acid; OVX, ovariectomy.

• 

  Fig. 4 Histological evaluation of ovariectomized rats with or without ASC injection. ASC injection during the early period (Week 1 and 2), late period (Week 5 and 6), or both after OVX. H&E staining was performed with subcutaneous fat and visceral fat. Arrows indicate subcutaneous fat layer. ASC, ascorbic acid; OVX, ovariectomy; H&E, hematoxylin and eosin.

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