Korean J Physiol Pharmacol.  2021 Jul;25(4):333-339. 10.4196/kjpp.2021.25.4.333.

The beneficial effect of glycerophosphocholine to local fat accumulation: a comparative study with phosphatidylcholine and aminophylline

• Kim GW
• Chung SH

• KMID: 2516872
• DOI: http://doi.org/10.4196/kjpp.2021.25.4.333

Abstract

Injection lipolysis or mesotherapy gained popularity for local fat dissolve as an alternative to surgical liposuction. Phosphatidylcholine (PPC) and aminophylline (AMPL) are commonly used compounds for mesotherapy, but their efficacy and safety as lipolytic agents have been controversial. Glycerophosphocholine (GPC) is a choline precursor structurally similar to PPC, and thus introduced in aesthetics as an alternative for PPC. This study aimed to evaluate the effects of GPC on adipocytes differentiation and lipolysis and compared those effects with PPC and AMPL using in vitro and in vivo models. Adipogenesis in 3T3-L1 was measured by Oil Red O staining. Lipolysis was assessed by measuring the amount of glycerol released in the culture media. To evaluate the lipolytic activity of GPC on a physiological condition, GPC was subcutaneously injected to one side of inguinal fat pads for 3 days. Lipolytic activity of GPC was assessed by hematoxylin and eosin staining in adipose tissue. GPC significantly suppressed adipocyte differentiation of 3T3-L1 in a concentration-dependent manner (22.3% inhibition at 4 mM of GPC compared to control). Moreover, when lipolysis was assessed by glycerol release in 3T3-L1 adipocytes, 6 mM of GPC stimulated glycerol release by two-fold over control. Subcutaneous injection of GPC into the inguinal fat pad of mice significantly reduced the mass of fat pad and the size of adipocytes of injected site, and these effects of GPC were more prominent over PPC and AMPL. Taken together, these results suggest that GPC is the potential therapeutic agent as a local fat reducer.

Keyword

Aminophylline; Choline alfoscerate; Glycerophosphocholine; Lipolysis; Mesotherapy; Phosphatidylcholine

Figure

• 

  Fig. 1 Effect of GPC on 3T3-L1 adipocyte viability. 3T3-L1 adipocytes were incubated with (A) GPC, (B) PPC and (C) AMPL at indicated concentrations for 96 h. Cell viability was determined by MTS incorporation-based cell proliferation assay. Assays were performed in triplicate for each treatment. (D) Chemical structure of experimental compounds is shown. GPC, glycerophosphocholine; PPC, phosphatidylcholine; AMPL, aminophylline. **p < 0.01 vs. control.

• 

  Fig. 2 Effect of GPC on lipid accumulation and lipolysis in 3T3-L1 adipocytes. (A) Representative photomicrographs show lipid accumulation in the cell treated with GPC. 3T3-L1 cells were induced to differentiate with adipogenic cocktail containing IBMX, dexamethasone and insulin (MDI) in the presence or absence of GPC, PPC or AMPL. Lipid accumulation was visualized with Oil Red O staining. Scale = 100 μm (B) Intracellular lipid content in differentiated adipocytes concomitantly treated with different compounds was quantified by elution of Oil Red O stain. ***p < 0.001 vs. MDI. (C) Differentiated 3T3-L1 adipocytes were incubated for 3 h in the absence of presence of GPC, PPC or AMPL. Lipolysis was quantified by glycerol release in incubation medium. Isoproterenol (Isop) was used as a positive control. *p < 0.05, **p < 0.01, ***p < 0.001 vs. non-treated cells. GPC, glycerophosphocholine; PPC, phosphatidylcholine; AMPL, aminophylline.

• 

  Fig. 3 Effect of GPC on body weight and food intake in DIO mice. (A) Change in body weight of the mice injected with GPC, PPC or AMPL was measured daily. Injection was initiated on day 0 and mice were sacrificed on day 3. (B) Body weight loss after treatment was calculated. (C) Change in food intake of mice injected with GPC, PPC or AMPL was measured daily. (D) Cumulative food intake during injection period was calculated. GPC, glycerophosphocholine; PPC, phosphatidylcholine; AMPL, aminophylline; DIO, diet-induced obesity. *p < 0.05, **p < 0.01, ***p < 0.001 vs. saline control.

• 

  Fig. 4 Effect of GPC on localized fat reduction. (A) Inguinal adipose tissue was dissected and measured after 3 days of GPC, PPC or AMPL injection. *p < 0.05 vs. saline control. (B) Ratio of injected or non-injected mouse inguinal fat pads mass was calculated. **p < 0.01 vs. saline control. (C) Inguinal adipose tissue section was prepared from injection site and stained by H&E. Scale bars indicate 100 μm. (D) Adipocyte area was measured, and the distribution of adipocyte is shown in each size range. GPC, glycerophosphocholine; PPC, phosphatidylcholine; AMPL, aminophylline.

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