Sulforaphane and myricetin act synergistically to induce apoptosis in 3T3‑L1 adipocytes

Yao,Anjun; Shen,Yingzhuo; Zhang,Zhuangwei; Zou,Zuquan; Wang,Anshi; Chen,Shiyong; Zhang,Huiqin; Chen,Fen; Zhao,Jinshun; Chen,Zhongming; Shan,Yujuan; Zhang,Xiaohong

doi:10.3892/mmr.2017.8235

Sulforaphane and myricetin act synergistically to induce apoptosis in 3T3‑L1 adipocytes

Authors:
- Anjun Yao
- Yingzhuo Shen
- Zhuangwei Zhang
- Zuquan Zou
- Anshi Wang
- Shiyong Chen
- Huiqin Zhang
- Fen Chen
- Jinshun Zhao
- Zhongming Chen
- Yujuan Shan
- Xiaohong Zhang
View Affiliations

Affiliations: Institute of Preventative Medicine, School of Medicine, Ningbo University, Ningbo, Zhejiang 315211, P.R. China, The Affiliated Hospital of School of Medicine of Ningbo University, Ningbo, Zhejiang 315210, P.R. China, Ningbo Kangning Hospital, Ningbo, Zhejiang 315210, P.R. China, School of Food Science and Engineering, Harbin Institute of Technology, Harbin, Heilongjiang 150090, P.R. China
Published online on: December 11, 2017 https://doi.org/10.3892/mmr.2017.8235
Pages: 2945-2951
Copyright: © Yao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate whether sulforaphane (SFN) and myricetin (Myr) synergistically induce apoptosis in adipocytes. The viability of mature 3T3‑L1 adipocytes treated with 40 µM SFN and/or 100 µM Myr was assessed using an MTT assay. Apoptosis was assessed by Hoechst 33258 nuclear staining, and by detection of single‑stranded DNA using an enzyme‑linked immunosorbent assay. Compared with the effects of each compound alone, the combination of SFN and Myr synergistically reduced cell viability, induced apoptosis, increased pro‑apoptotic Bcl‑2 associated X protein expression, decreased anti‑apoptotic B‑cell lymphoma‑2 expression, enhanced Bcl‑2‑associated death promoter (Bad) translocation from the cytoplasm to the mitochondria, and reduced Bad phosphorylation at Ser112. These effects were accompanied by increased cleavage of caspase 3 and poly‑ADP‑ribose‑polymerase. In addition, combined SFN and Myr treatment significantly decreased the protein expression levels of phosphorylated AKT serine/threonine kinase 1 (Akt) at Ser473, as well as the phosphorylation of the downstream protein ribosomal protein, S6 kinase β‑1. Therefore, SFN plus Myr was a more potent inducer of apoptosis in 3T3‑L1 adipocytes than either compound alone. The results of the present study suggest that the mechanism of SNF/Myr‑induced apoptosis involved activation of the Akt‑mediated mitochondrial apoptotic pathway. This may aid treatment of animal models of obesity and preclinical testing.

View Figures

View References

1	World Health Organization, . Fact sheet no 311. 2015, http://www.who.int/mediacentre/factsheets/fs311/en/print.htmlDec. 4–2015
2	Jo J, Gavrilova O, Pack S, Jou W, Mullen S, Sumner AE, Cushman SW and Periwal V: Hypertrophy and/or Hyperplasia: Dynamics of adipose tissue growth. PLoS Comput Biol. 5:e10003242009. View Article : Google Scholar : PubMed/NCBI
3	Prins JB, Walker NI, Winterford CM and Cameron DP: Human adipocyte apoptosis occurs in malignancy. Biochem Biophys Res Commun. 205:625–630. 1994. View Article : Google Scholar : PubMed/NCBI
4	Prins JB, Walker NI, Winterford CM and Cameron DP: Apoptosis of human adipocytes in vitro. Biochem Biophys Res Commun. 201:500–507. 1994. View Article : Google Scholar : PubMed/NCBI
5	Lee YJ and Lee SH: Sulforaphane induces antioxidative and antiproliferative responses by generating reactive oxygen species in human bronchial epithelial BEAS-2B cells. J Korean Med Sci. 26:1474–1482. 2011. View Article : Google Scholar : PubMed/NCBI
6	Juengel E, Maxeiner S, Rutz J, Justin S, Roos F, Khoder W, Tsaur I, Nelson K, Bechstein WO, Haferkamp A and Blaheta RA: Sulforaphane inhibits proliferation and invasive activity of everolimus-resistant kidney cancer cells in vitro. Oncotarget. 7:85208–85219. 2016.PubMed/NCBI
7	Shehatou GS and Suddek GM: Sulforaphane attenuates the development of atherosclerosis and improves endothelial dysfunction in hypercholesterolemic rabbits. Exp Biol Med (Maywood). 241:426–436. 2016. View Article : Google Scholar : PubMed/NCBI
8	Li Y, Zhang T, Korkaya H, Liu S, Lee HF, Newman B, Yu Y, Clouthier SG, Schwartz SJ, Wicha MS and Sun D: Sulforaphane, a dietary component of broccoli/broccoli sprouts, inhibits breast cancer stem cells. Clin Cancer Res. 16:2580–2590. 2010. View Article : Google Scholar : PubMed/NCBI
9	Sarkar R, Mukherjee S, Biswas J and Roy M: Sulphoraphane, a naturally occurring isothiocyanate induces apoptosis in breast cancer cells by targeting heat shock proteins. Biochem Biophys Res Commun. 427:80–85. 2012. View Article : Google Scholar : PubMed/NCBI
10	Choi KM, Lee YS, Kim W, Kim SJ, Shin KO, Yu JY, Lee MK, Lee YM, Hong JT, Yun YP and Yoo HS: Sulforaphane attenuates obesity by inhibiting adipogenesis and activating the AMPK pathway in obese mice. J Nutr Biochem. 25:201–207. 2014. View Article : Google Scholar : PubMed/NCBI
11	Lee JH, Moon MH, Jeong JK, Park YG, Lee YJ, Seol JW and Park SY: Sulforaphane induced adipolysis via hormone sensitive lipase activation, regulated by AMPK signaling pathway. Biochem Biophys Res Commun. 426:492–497. 2012. View Article : Google Scholar : PubMed/NCBI
12	Choi KM, Lee YS, Sin DM, Lee S, Lee MK, Lee YM, Hong JT, Yun YP and Yoo HS: Sulforaphane inhibits mitotic clonal expansion during adipogenesis through cell cycle arrest. Obesity (Silver Spring). 20:1365–1371. 2012. View Article : Google Scholar : PubMed/NCBI
13	Yao A, Shen Y, Wang A, Chen S, Zhang H, Chen F, Chen Z, Wei H, Zou Z, Shan Y and Zhang X: Sulforaphane induces apoptosis in adipocytes via Akt/p70s6k1/Bad inhibition and ERK activation. Biochem Biophys Res Commun. 465:696–701. 2015. View Article : Google Scholar : PubMed/NCBI
14	Ong KC and Khoo HE: Biological effects of myricetin. Gen Pharmacol. 29:121–126. 1997. View Article : Google Scholar : PubMed/NCBI
15	Ross JA and Kasum CM: Dietary flavonoids: Bioavailability, metabolic effects, and safety. Annu Rev Nutr. 22:19–34. 2002. View Article : Google Scholar : PubMed/NCBI
16	Weng CJ and Yen GC: Flavonoids, a ubiquitous dietary phenolic subclass, exert extensive in vitro anti-invasive and in vivo anti-metastatic activities. Cancer Metastasis Rev. 31:323–351. 2012. View Article : Google Scholar : PubMed/NCBI
17	Sun F, Zheng XY, Ye J, Wu TT, Wang J and Chen W: Potential anticancer activity of myricetin in human T24 bladder cancer cells both in vitro and in vivo. Nutr Cancer. 64:599–606. 2012. View Article : Google Scholar : PubMed/NCBI
18	Phillips PA, Sangwan V, Borja-Cacho D, Dudeja V, Vickers SM and Saluja AK: Myricetin induces pancreatic cancer cell death via the induction of apoptosis and inhibition of the phosphatidylinositol 3-kinase (PI3K) signaling pathway. Cancer Lett. 308:181–188. 2011. View Article : Google Scholar : PubMed/NCBI
19	Datta SR, Brunet A and Greenberg ME: Cellular survival: A play in three Akts. Genes Dev. 13:2905–2927. 1999. View Article : Google Scholar : PubMed/NCBI
20	Bonni A, Brunet A, West AE, Datta SR, Takasu MA and Greenberg ME: Cell survival promoted by the Ras-MAPK signaling pathway by transcription-dependent and -independent mechanisms. Science. 286:1358–1362. 1999. View Article : Google Scholar : PubMed/NCBI
21	Chang CJ, Tzeng TF, Liou SS, Chang YS and Liu IM: Myricetin increases hepatic peroxisome proliferator-activated receptor α protein expression and decreases plasma lipids and adiposity in rats. Evid Based Complement Alternat Med. 2012:7871522012. View Article : Google Scholar : PubMed/NCBI
22	Wang Q, Wang ST, Yang X, You PP and Zhang W: Myricetin suppresses differentiation of 3 T3-L1 preadipocytes and enhances lipolysis in adipocytes. Nut Rese. 35:317–327. 2015. View Article : Google Scholar
23	Yang JY, Della-Fera MA, Rayalam S and Baile CA: Enhanced effects of xanthohumol plus honokiol on apoptosis in 3T3-L1 adipocytes. Obesity (Silver Spring, Md). 16:1232–1238. 2008. View Article : Google Scholar : PubMed/NCBI
24	Yang JY, Della-Fera MA, Rayalam S, Ambati S, Hartzell DL, Park HJ and Baile CA: Enhanced inhibition of adipogenesis and induction of apoptosis in 3T3-L1 adipocytes with combinations of resveratrol and quercetin. Life Sci. 82:1032–1039. 2008. View Article : Google Scholar : PubMed/NCBI
25	Rayalam S, Yang JY, Della-Fera MA, Park HJ, Ambati S and Baile CA: Anti-obesity effects of xanthohumol plus guggulsterone in 3T3-L1 adipocytes. J Med Food. 12:846–853. 2009. View Article : Google Scholar : PubMed/NCBI
26	Baile CA, Yang JY, Rayalam S, Hartzell DL, Lai CY, Andersen C and Della-Fera MA: Effect of resveratrol on fat mobilization. Ann N Y Acad Sci. 1215:40–47. 2011. View Article : Google Scholar : PubMed/NCBI
27	Porter AG and Jänicke RU: Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 6:99–104. 1999. View Article : Google Scholar : PubMed/NCBI
28	Rayalam S, Della-Fera MA and Baile CA: Phytochemicals and regulation of the adipocyte life cycle. J Nutr Biochem. 19:717–726. 2008. View Article : Google Scholar : PubMed/NCBI
29	Rayalam S, Della-Fera MA, Yang JY, Park HJ, Ambati S and Baile CA: Resveratrol potentiates genistein's antiadipogenic and proapoptotic effects in 3T3-L1 adipocytes. J Nutr. 137:2668–2673. 2007.PubMed/NCBI
30	Vander Heiden MG and Thompson CB: Bcl-2 proteins: Regulators of apoptosis or of mitochondrial homeostasis? Nat Cell Biol. 1:E209–E216. 1999. View Article : Google Scholar : PubMed/NCBI
31	Siddiqui WA, Ahad A and Ahsan H: The mystery of BCL2 family: Bcl-2 proteins and apoptosis: An update. Arch Toxicol. 89:289–317. 2015. View Article : Google Scholar : PubMed/NCBI
32	Hsu CL and Yen GC: Effects of capsaicin on induction of apoptosis and inhibition of adipogenesis in 3T3-L1 cells. J Agric Food Chem. 55:1730–1736. 2007. View Article : Google Scholar : PubMed/NCBI
33	Zhu L, Han MB, Gao Y, Wang H, Dai L, Wen Y and Na LX: Curcumin triggers apoptosis via upregulation of Bax/Bcl-2 ratio and caspase activation in SW872 human adipocytes. Mol Med Rep. 12:1151–1156. 2015. View Article : Google Scholar : PubMed/NCBI
34	Agarwal A, Mahfouz RZ, Sharma RK, Sarkar O, Mangrola D and Mathur PP: Potential biological role of poly (ADP-ribose) polymerase (PARP) in male gametes. Reprod Biol Endocrinol. 7:1432009. View Article : Google Scholar : PubMed/NCBI
35	Jakubíková J, Sedlák J, Mithen R and Bao Y: Role of PI3K/Akt and MEK/ERK signaling pathways in sulforaphane- and erucin-induced phase II enzymes and MRP2 transcription, G2/M arrest and cell death in Caco-2 cells. Biochem Pharmacol. 69:1543–1552. 2005. View Article : Google Scholar : PubMed/NCBI
36	Zhang XH, Chen SY, Tang L, Shen YZ, Luo L, Xu CW, Liu Q and Li D: Myricetin induces apoptosis in HepG2 cells through Akt/p70S6K/bad signaling and mitochondrial apoptotic pathway. Anticancer Agents Med Chem. 13:1575–1581. 2013. View Article : Google Scholar : PubMed/NCBI