Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Identification of anti-adipogenic withanolides from the roots of Indian ginseng (Withania somnifera)

  • Lee, Seoung Rak (School of Pharmacy, Sungkyunkwan University) ;
  • Lee, Bum Soo (School of Pharmacy, Sungkyunkwan University) ;
  • Yu, Jae Sik (School of Pharmacy, Sungkyunkwan University) ;
  • Kang, Heesun (School of Pharmacy, Sungkyunkwan University) ;
  • Yoo, Min Jeong (School of Pharmacy, Sungkyunkwan University) ;
  • Yi, Sang Ah (School of Pharmacy, Sungkyunkwan University) ;
  • Han, Jeung-Whan (School of Pharmacy, Sungkyunkwan University) ;
  • Kim, Sil (School of Pharmacy, Sungkyunkwan University) ;
  • Kim, Jung Kyu (School of Chemical Engineering, Sungkyunkwan University) ;
  • Kim, Jin-Chul (KIST Gangneung Institute of Natural Products, Natural Product Informatics Research Center) ;
  • Kim, Ki Hyun (School of Pharmacy, Sungkyunkwan University)
  • Received : 2021.02.04
  • Accepted : 2021.09.30
  • Published : 2022.05.01
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Abstract

Background: Withania somnifera (Solanaceae), generally known as Indian ginseng, is a medicinal plant that is used in Ayurvedic practice for promoting health and longevity. This study aims to identify the bioactive metabolites from Indian ginseng and elucidate their structures. Methods: Withanolides were purified by chromatographic techniques, including HPLC coupled with LC/MS. Chemical structures of isolated withanolides were clarified by analyzing the spectroscopic data from 1D and 2D NMR, and HR-ESIMS experiment. Absolute configurations of the withanolides were established by the application of NMR chemical shifts and ECD calculations. Anti-adipogenic activities of isolates were evaluated using 3T3-L1 preadipocytes with Oil Red O staining and quantitative real-time PCR (qPCR). Results: Phytochemical examination of the roots of Indian ginseng afforded to the isolation of six withanolides (1-6), including three novel withanolides, withasilolides GeI (1-3). All the six compounds inhibited adipogenesis and suppressed the enlargement of lipid droplets, compared to those of the control. Additionally, the mRNA expression levels of Fabp4 and Adipsin, the adipocyte markers decreased noticeably following treatment with 25 µM of 1-6. The active compounds (1-6) also promoted lipid metabolism by upregulating the expression of the lipolytic genes HSL and ATGL and downregulating the expression of the lipogenic gene SREBP1. Conclusion: The results of our experimental studies suggest that the withasilolides identified herein have anti-adipogenic potential and can be considered for the development of therapeutic strategies against adipogenesis in obesity. Our study also provides a mechanistic rationale for using Indian ginseng as a potential therapeutic agent against obesity and related metabolic diseases.

Keywords

Acknowledgement

This work was supported by a grant from the National Research Foundation of Korea (NRF), funded by the Korean government (MSIT) (grant number: 2019R1A5A2027340 and 2021R1A2C2007937).

References

  1. Jayaprakasam B, Zhang Y, Seeram NP, Nair MG. Growth inhibition of human tumor cell lines by withanolides from Withania somnifera leaves. Life Sci 2003;74:125-32. https://doi.org/10.1016/j.lfs.2003.07.007
  2. Choudhary MI, Yousuf S, Nawaz SA, Ahmed S, Atta R. Cholinesterase inhibiting withanolides from Withania somnifera. Chem Pharm Bull 2004;52:1358-61. https://doi.org/10.1248/cpb.52.1358
  3. Mary NK, Babu BH, Padikkala J. Antiatherogenic effect of Caps HT2, a herbal Ayurvedic medicine formulation. Phytomedicine 2003;10:474-82. https://doi.org/10.1078/094471103322331412
  4. Gupta GL, Rana AC. PHCOG MAG.: plant review Withania somnifera (Ashwagandha): a review. Phcog Rev 2007;1:129-36.
  5. Srivastava A, Gupta AK, Shanker K, Gupta MM, Mishra R, Lal RK. Genetic variability, associations, and path analysis of chemical and morphological traits in Indian ginseng [Withania somnifera (L.) Dunal] for selection of higher yielding genotypes. J Ginseng Res 2018;42:128-64.
  6. Ganguly B, Kumar N, Ahmad AH, Rastogi SK. Influence of phytochemical composition on in vitro antioxidant and reducing activities of Indian ginseng [Withania somnifera (L.) Dunal] root extracts. J Ginseng Res 2018;42:463-9. https://doi.org/10.1016/j.jgr.2017.05.002
  7. Singh S, Kumar S. Withania somnifera. The Indian ginseng ashwagandha. Lucknow, India: Central Institute of Medicinal and Aromatic Plants; 1998.
  8. Atta-ur-Rahman YM, Gul W, Qureshi S, Choudhary MI, Voelter W, Hoff A, Jens F, Naz A. Cholinesterase inhibiting withanolides from Withania somnifera. Heterocycles 1998;48:1801-11. https://doi.org/10.3987/COM-98-8208
  9. Bhattacharya SK, Satyan KS, Chakrabarti A. Effect of Trasina, an Ayurvedic herbal formulation, on pancreatic islet superoxide dismutase activity in hyperglycaemic rats. Indian J Exp Biol 1997;35:297-9.
  10. Dhuley JN. Effect of ashwagandha on lipid peroxidation in stress-induced animals. J Ethnopharmacol 1998;60:173-8. https://doi.org/10.1016/S0378-8741(97)00151-7
  11. Bhattacharya A, Ramanathan M, Ghosal S, Bhattacharya SK. Effect of Withania somnifera glycowithanolides on iron-induced hepatotoxicity in rats. Phytother Res 2000;14:568-70. https://doi.org/10.1002/1099-1573(200011)14:7<568::AID-PTR663>3.0.CO;2-Q
  12. Kulkarni SK, Verma A. Prevention of development of tolerance and dependence to opiate in mice by BR-16A (Mentat ®), a herbal psychotropic preparation. Indian J Exp Biol 1992;30:885-8.
  13. Ganzera M, Choudhary MI, Khan IA. Quantitative HPLC analysis of withanolides in Withania somnifera. Fitoterapia 2003;74:68-76. https://doi.org/10.1016/S0367-326X(02)00325-8
  14. Elsakka M, Grigorescu E, Stanescu U, Stanescu U, Dorneanu V. New data referring to chemistry of Withania somnifera species. Rev Med-Chir Soc Med Nat Iasi 1990;94:385-7.
  15. Matsuda H, Murakami T, Kishi A, Yoshikawa M. Structures of withanosides I, II, III, IV, V, VI and VII new withanolide glycosides from the roots of Indian Withania somnifera D and inhibitory activity for tachyphylaxis to clonidine in isolated guineapig ileum. Bioorg Med Chem 2001;96:1499-507.
  16. Davis L, Kuttan G. Immunomodulatory activity of Withania somnifera. J Ethnopharmacol 2000;71:193-200. https://doi.org/10.1016/S0378-8741(99)00206-8
  17. Park HJ, Rayalam S, Della-Fera MA, Ambati S, Yang JY, Baile CA. Withaferin A induces apoptosis and inhibits adipogenesis in 3T3-L1 adipocytes. Biofactors 2008;33:137-48. https://doi.org/10.1002/biof.5520330206
  18. Lee S, Ryoo R, Choi JH, Kim JH, Kim SH, Kim KH. Trichothecene and tremulane sesquiterpenes from a hallucinogenic mushroom Gymnopilus junonius and their cytotoxicity. Arch Pharm Res (Seoul) 2020;43:214-23. https://doi.org/10.1007/s12272-020-01213-6
  19. Ha JW, Kim J, Kim H, Jang W, Kim KH. Mushrooms: an important source of natural bioactive compounds. Nat Prod Sci 2020;26:118-31. https://doi.org/10.20307/nps.2020.26.2.118
  20. Lee SR, Kang H, Yoo MJ, Yu JS, Lee S, Yi SA, Beemelmanns C, Lee J, Kim KH. Anti-adipogenic pregnane steroid from a Hydractinia-associated fungus, Cladosporium sphaerospermum SW67. Nat Prod Sci 2020;26:230-5. https://doi.org/10.20307/nps.2020.26.3.230
  21. Yu JS, Park M, Pang C, Rashan L, Jung WH, Kim KH. Antifungal phenols from Woodfordia uniflora collected in Oman. J Nat Prod 2020;83:2261-8. https://doi.org/10.1021/acs.jnatprod.0c00395
  22. Kim S, Yu JS, Lee JY, Choi SU, Lee J, Kim KH. Cytotoxic withanolides from the roots of Indian ginseng (Withania somnifera). J Nat Prod 2019;82:765-73. https://doi.org/10.1021/acs.jnatprod.8b00665
  23. Baek SC, Lee S, Kim S, Jo MS, Yu JS, Ko YJ, Cho YC, Kim KH. Withaninsams A and B: phenylpropanoid esters from the roots of Indian ginseng (Withania somnifera). Plants 2019;8:527. https://doi.org/10.3390/plants8120527
  24. Yu JS, Li C, Kwon M, Oh T, Lee TH, Kim DH, Ahn JS, Ko SK, Kim CS, Cao S, Kim KH, Herqueilenone A. A unique rearranged benzoquinone-chromanone from the Hawaiian volcanic soil-associated fungal strain Penicillium herquei FT729. Bioorg Chem 2020;105:104397. https://doi.org/10.1016/j.bioorg.2020.104397
  25. Lee SR, Lee D, Park M, Lee JC, Park HJ, Kang KS, Kim CE, Beemelmanns C, Kim KH. Absolute configuration and corrected NMR assignment of 17-hydroxycyclooctatin, a fused 5-8-5 tricyclic diterpene. J Nat Prod 2020;83:354-61. https://doi.org/10.1021/acs.jnatprod.9b00837
  26. Kuroyanagi M, Shibata K, Umehara K. Cell differentiation inducing steroids from Withania somnifera L.(Dun.). Chem Pharm Bull 1999;47:1646-9. https://doi.org/10.1248/cpb.47.1646
  27. Xu YM, Wijeratne EMK, Brooks AD, Tewary P, Xuan LJ, Wang WQ, Sayers TJ, Gunatilaka AAL. Cytotoxic and other withanolides from aeroponically grown Physalis philadelphica. Phytochemistry 2018;152:174-81. https://doi.org/10.1016/j.phytochem.2018.04.018
  28. Smith SG, Goodman JM. Assigning the stereochemistry of pairs of diastereoisomers using GIAO NMR shift calculation. J Org Chem 2009;74:4597-607. https://doi.org/10.1021/jo900408d
  29. Smith SG, Goodman JM. Assigning stereochemistry to single diastereoisomers by GIAO NMR calculation: the DP4 probability. J Am Chem Soc 2010;132:12946-59. https://doi.org/10.1021/ja105035r
  30. Hehre W, Klunzinger P, Deppmeier B, Driessen A, Uchida N, Hashimoto M, Fukushi E, Takata Y. Efficient protocol for accurately calculating 13C chemical shifts of conformationally flexible natural products: scope, assessment, and limitations. J Nat Prod 2019;82:2299-306. https://doi.org/10.1021/acs.jnatprod.9b00603
  31. Spiegelman BM, Flier JS. Obesity and the regulation of energy balance. Cell 2001;104:531-43. https://doi.org/10.1016/S0092-8674(01)00240-9
  32. Smith U, Kahn BB. Adipose tissue regulates insulin sensitivity: role of adipogenesis, de novo lipogenesis and novel lipids. J Intern Med 2016;280:465-75. https://doi.org/10.1111/joim.12540
  33. Lee K, Seo YJ, Song JH, Chei S, Lee BY. Ginsenoside Rg1 promotes browning by inducing UCP1 expression and mito-chondrial activity in 3T3-L1 and subcutaneous white adipocytes. J Ginseng Res 2019;43:589-99. https://doi.org/10.1016/j.jgr.2018.07.005
  34. Yi SA, Lee J, Park SK, Kim JY, Park JW, Lee MG, Nam KH, Park JH, Oh H, Kim S, Han J, Kim BK, Jo DG, Han JW. Fermented ginseng extract, BST204, disturbs adipogenesis of mesenchymal stem cells through inhibition of S6 kinase 1 signaling. J Ginseng Res 2020;44:58-66. https://doi.org/10.1016/j.jgr.2018.08.002