Journal List > J Nutr Health > v.52(1) > 1117285

J Nutr Health. 2019 Feb;52(1):17-25. Korean.
Published online Feb 28, 2019.
© 2019 The Korean Nutrition Society
Effects of Lonicera caerulea extract on adipocyte differentiation and adipogenesis in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs)
Miey Park,1 Changho Lee,2 and Hae-Jeung Lee1
1Department of Food and Nutrition, Gachon University, Seongnam, Gyeonggi 13120, Korea.
2Research Group of Functional Food Materials, Korea Food Research Institute, Wanju, Jeonbuk 55365, Korea.

To whom correspondence should be addressed. tel: +82-31-750-5968, Email: ,Email:
Received Jan 09, 2019; Revised Jan 18, 2019; Accepted Feb 09, 2019.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.



Obesity is a major health problem of global significance because it is clearly associated with an increased risk of health problems, such as nonalcoholic fatty liver disease (NAFLD), diabetes, cardiovascular diseases, and cancer. Lonicera caerulea (LC) originates from high mountains or wet areas and has been used as a traditional medicine in northern Russia, China, and Japan. LC contains a range of bioactive constituents, such as vitamins, minerals, and polyphenols. This study examined the anti-obesity effects of LC during differentiation in preadipocytes.


The cell viability assay was performed after the differentiation of 3T3-L1 cells for 7 days. Oil Red O staining was used to visualize the changes in lipid droplets in 3T3-L1 cells and mouse adipose-derived stem cells (MADSCs). The mRNA expression of obesity-related genes was determined by quantitative real-time PCR.


According to the results of Oil Red O staining, the lipid levels and size of lipid droplets in the adipocytes were reduced and the LC extract (LCE, 0.25–1 mg/mL) markedly inhibited adipogenesis in a dose-dependent manner. The treatment of LCE also decreased the mRNA expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα), and sterol regulatory element binding protein 1 (SREBP1) in 3T3-L1 cells. Western blot analysis showed that the PPARγ, C/EBPα, and SREBP1 protein levels in both 3T3-L1 and MADSC were reduced in a dose-dependent manner.


These results suggest that LCE can inhibit adipogenic differentiation through the regulation of adipogenesis-related markers.

Keywords: Lonicera caerulea; preadipocyte; anti-obesity; lipid accumulation


Fig. 1
Effect of Lonicera caerulea extract (LCE) on 3T3-L1 preadipocytes proliferation. 3T3-L1 preadipocyte cells were treated with the indicated concentrations of LCE for 24, 48, and 72hr and cell viability was estimated by a CCK-8 assay. All experiments were repeated at least three times and data represent means ± SD. ** p < 0.01, *** p < 0.001 vs. Con.
Click for larger image

Fig. 2
Lipid accumulation was determined by Oil Red O staining. LCE treated 3T3-L1 adipocytes (A) and lipid levels in 3T3-L1 adipocytes (B). Original magnification 200X. Data represent means ± SD. ** p < 0.01, *** p < 0.001 vs. Con.
Click for larger image

Fig. 3
Lipid accumulation was determined by Oil Red O staining. LCE treated mouse adipose-derived stem cell (MADSC) adipocytes (A) and lipid levels in MADSC adipocytes (B). Original magnification 200X. Data represent means ± SD. *** p < 0.001 vs. Con.
Click for larger image

Fig. 4
Effects of LCE on the expression of genes associated with adipogenesis in 3T3-L1 cells. The expression of Pparγ (A), C/ebpα (B), and Srebp1 (C) were quantified by real-time PCR and normalized by β-actin as an internal control. (D) Western blot analysis of PPARγ, C/EBPα, and SREBP1 protein. Data represent means ± SD. * p < 0.05, ** p < 0.01 vs. Con.
Click for larger image

Fig. 5
Effects of LCE on the expression of protein associated with adipogenesis in MADSCs. The protein expression of PPARγ (A), C/EBPα (B), and SREBP1 (C) were normalized by β-actin as an internal control. (D) PPARγ, C/EBPα, and SREBP1 protein levels by Immunoblot analysis. Data represent means ± SD. * p < 0.05, ** p < 0.01 vs. Con.
Click for larger image


Table 1
Sequences of primers for quantitative real-time PCR in this research
Click for larger image


This work was supported by a grant ‘Cooperative Research Program for Agriculture Science and Technology Development funded by Rural Development Administration (Project No. PJ01227802) and Korea Food Research Institute (grant E0150302-04), Republic of Korea’.

1. Yatsuya H, Li Y, Hilawe EH, Ota A, Wang C, Chiang C, et al. Global trend in overweight and obesity and its association with cardiovascular disease incidence. Circ J 2014;78(12):2807–2818.
2. Kopelman PG. Obesity as a medical problem. Nature 2000;404(6778):635–643.
3. Must A, Spadano J, Coakley EH, Field AE, Colditz G, Dietz WH. The disease burden associated with overweight and pathway. Toxicol Res 2018;34(1):13–21.
4. Bell L, Lamport DJ, Butler LT, Williams CM. A review of the cognitive effects observed in humans following acute supplementation with flavonoids, and their associated mechanisms of action. Nutrients 2015;7(12):10290–10306.
5. Han MH, Kim HJ, Jeong JW, Park C, Kim BW, Choi YH. Inhibition of adipocyte differentiation by anthocyanins isolated from the fruit of Vitis coignetiae pulliat is associated with the activation of AMPK signaling pathway. Toxicol Res 2018;34(1):13–21.
6. Leu SY, Chen YC, Tsai YC, Hung YW, Hsu CH, Lee YM, et al. Raspberry ketone reduced lipid accumulation in 3T3-L1 cells and ovariectomy-induced obesity in wistar rats by regulating autophagy mechanisms. J Agric Food Chem 2017;65(50):10907–10914.
7. Rupasinghe HP, Yu LJ, Bhullar KS, Bors B. Short communication: haskap (Lonicera caerulea): a new berry crop with high antioxidant capacity. Can J Plant Sci 2012;92(7):1311–1317.
8. Kähkönen MP, Hopia AI, Heinonen M. Berry phenolics and their antioxidant activity. J Agric Food Chem 2001;49(8):4076–4082.
9. Wang SY, Lin HS. Antioxidant activity in fruits and leaves of blackberry, raspberry, and strawberry varies with cultivar and developmental stage. J Agric Food Chem 2000;48(2):140–146.
10. Han SJ. Protective efficacies of Aronia melanocarpa (blackberry) on the allyl alcohol-damaged hepatocyte of mice. Korean J Pharmacogn 2013;44(1):91–96.
11. Kim JH, Jo MN, Han JH, Pyo SH, Kim TS, Kim EH, et al. Anti-inflammatory activity of the fruits of blue honeysuckle. Yakhak Hoeji 2016;60(5):235–242.
12. Ko GA, Koh YS, Ryu JY, Kim Cho S. Comparison of proximate compositions, antioxidant, and antiproliferative activities between blueberry and Sageretia thea (Osbeck) M.C. Johnst fruit produced in Jeju Island. J Appl Biol Chem 2017;60(2):161–171.
13. Zhao H, Wang Z, Ma F, Yang X, Cheng C, Yao L. Protective effect of anthocyanin from Lonicera caerulea var. edulis on radiation-induced damage in mice. Int J Mol Sci 2012;13(9):11773–11782.
14. Vostálová J, Galandáková A, Palíková I, Ulrichová J, Doležal D, Lichnovská R, et al. Lonicera caerulea fruits reduce UVA-induced damage in hairless mice. J Photochem Photobiol B 2013;128:1–11.
15. Aune UL, Ruiz L, Kajimura S. Isolation and differentiation of stromal vascular cells to beige/brite cells. J Vis Exp 2013;(73):50191.
16. Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, et al. Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med 2005;54(3):132–141.
17. Harmon AW, Harp JB. Differential effects of flavonoids on 3T3-L1 adipogenesis and lipolysis. Am J Physiol Cell Physiol 2001;280(4):C807–C813.
18. Rosen ED, MacDougald OA. Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol 2006;7(12):885–896.
19. Lee YM, Yoon Y, Yoon H, Park HM, Song S, Yeum KJ. Dietary anthocyanins against obesity and inflammation. Nutrients 2017;9(10):pii: E1089.
20. Azzini E, Giacometti J, Russo GL. Antiobesity effects of anthocyanins in preclinical and clinical studies. Oxid Med Cell Longev 2017;2017:2740364
21. Lee B, Lee M, Lefevre M, Kim HR. Anthocyanins inhibit lipogenesis during adipocyte differentiation of 3T3-L1 preadipocytes. Plant Foods Hum Nutr 2014;69(2):137–141.
22. Tontonoz P, Hu E, Spiegelman BM. Regulation of adipocyte gene expression and differentiation by peroxisome proliferator activated receptor γ. Curr Opin Genet Dev 1995;5(5):571–576.
23. Wu Z, Puigserver P, Spiegelman BM. Transcriptional activation of adipogenesis. Curr Opin Cell Biol 1999;11(6):689–694.
24. Oh SW. Recent epidemiological changes in Korean obesity. Korean J Helicobacter Up Gastrointest Res 2017;17(2):62–65.
25. Muir LA, Neeley CK, Meyer KA, Baker NA, Brosius AM, Washabaugh AR, et al. Adipose tissue fibrosis, hypertrophy, and hyperplasia: correlations with diabetes in human obesity. Obesity (Silver Spring) 2016;24(3):597–605.
26. Ailhaud G, Grimaldi P, Négrel R. Cellular and molecular aspects of adipose tissue development. Annu Rev Nutr 1992;12(1):207–233.
27. MacDougald OA, Hwang CS, Fan H, Lane MD. Regulated expression of the obese gene product (leptin) in white adipose tissue and 3T3-L1 adipocytes. Proc Natl Acad Sci U S A 1995;92(20):9034–9037.
28. Yu S, Shin S. Effects of Carthamus tinctorius extract on adipogenic differentiation of mouse bone marrow-derived mesenchymal stromal stem cells. J Int Korean Med 2017;38(1):1–9.
29. Chen G, Li H, Zhao Y, Zhu H, Cai E, Gao Y, et al. Saponins from stems and leaves of Panax ginseng prevent obesity via regulating thermogenesis, lipogenesis and lipolysis in high-fat diet-induced obese C57BL/6 mice. Food Chem Toxicol 2017;106(Pt A):393–403.
30. Zuk PA. The adipose-derived stem cell: looking back and looking ahead. Mol Biol Cell 2010;21(11):1783–1787.
31. Grégoire F, Todoroff G, Hauser N, Remacle C. The stroma-vascular fraction of rat inguinal and epididymal adipose tissue and the adipoconversion of fat cell precursors in primary culture. Biol Cell 1990;69(3):215–222.
32. Pandurangan M, Park J, Kim E. Aspartame downregulates 3T3-L1 differentiation. In Vitro Cell Dev Biol Anim 2014;50(9):851–857.
33. Umek RM, Friedman AD, McKnight SL. CCAAT-enhancer binding protein: a component of a differentiation switch. Science 1991;251(4991):288–292.
34. Cornelius P, MacDougald OA, Lane MD. Regulation of adipocyte development. Annu Rev Nutr 1994;14(1):99–129.
35. Smolik M, Ochmian I, Grajkowski J. Genetic variability of Polish and Russian accessions of cultivated blue honeysuckle (Lonicera caerulea). Genetika 2010;46(8):1079–1085. [doi: 10.1134/s1022795410080077]
36. Plekhanova MN. Blue honeysuckle (Lonicera caerulea L.) - a new commercial berry crop for temperate climate: genetic resources and breeding. Acta Hortic 2000;538(538):159–164.
37. Skupień K, Ochmian I, Grajkowski J. Influence of ripening time on fruit chemical composition of two blue honeysuckle cultigens. J Fruit Ornam Plant Res 2009;17(1):101–111.
38. Joseph SV, Edirisinghe I, Burton-Freeman BM. Berries: anti-inflammatory effects in humans. J Agric Food Chem 2014;62(18):3886–3903.
39. Song Y, Park HJ, Kang SN, Jang SH, Lee SJ, Ko YG, et al. Blueberry peel extracts inhibit adipogenesis in 3T3-L1 cells and reduce high-fat diet-induced obesity. PLoS One 2013;8(7):e69925
40. Tsuda T. Recent progress in anti-obesity and anti-diabetes effect of berries. Antioxidants (Basel) 2016;5(2):pii: E13.
41. Kim HK, Kim JN, Han SN, Nam JH, Na HN, Ha TJ. Black soybean anthocyanins inhibit adipocyte differentiation in 3T3-L1 cells. Nutr Res 2012;32(10):770–777.
42. Khan MI, Shin JH, Shin TS, Kim MY, Cho NJ, Kim JD. Anthocyanins from Cornus kousa ethanolic extract attenuate obesity in association with anti-angiogenic activities in 3T3-L1 cells by down-regulating adipogeneses and lipogenesis. PLoS One 2018;13(12):e0208556