Phytochemical characterization, biochemical profiling and evaluation of anticancer potential of methanolic extract of Withania somnifera stem

Aditi Srivastava; Rumana Ahmad; Sahabjada Siddiqui; Sudhir Mehrotra; Mohsin A. Khan

doi:10.1515/jcim-2024-0185

Article

Phytochemical characterization, biochemical profiling and evaluation of anticancer potential of methanolic extract of Withania somnifera stem

Aditi Srivastava , Rumana Ahmad , Sahabjada Siddiqui , Sudhir Mehrotra and Mohsin A. Khan

Published/Copyright: August 20, 2024

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From the journal Journal of Complementary and Integrative Medicine Volume 22 Issue 2

Abstract

Objectives

Phytotherapy employs phytoconstituents/phytomedicines derived from plants for treating and preventing illnesses. Withania somnifera is known in the Indian Ayurveda Pharmacopoeia for its medicinal applications and pharmacological properties. In this study, we examined the biological activity spectrum of Withania somnifera methanolic extract of stem (WSME), which is valued as a “Rasayana” due to its extensive range of medicinal uses.

Methods

WSME was subjected to TPC and TFC quantification and bioactive components were characterized using LC-MS. Its antioxidant potential was gauged by DPPH and H₂O₂ radical scavenging assays, while antibacterial efficacy was assessed against S. aureus and E. coli using disc diffusion assay. In vitro anticancer activity was evaluated against human breast cancer MDA-MB-231 cells while toxicity was evaluated against normal Vero cells using MTT assay.

Results

WSME, rich in Withaferin A, showed TPC of 4.73 ± 0.15 mg GAE/g and TFC of 94.94 ± 6.15 mg QE/g dry weight of extract. It exhibited significant antioxidant activity (43.28 and 66.8 % inhibition at 1,000 μg/mL using DPPH and H₂O₂ assays, respectively) and mild antibacterial effects against S. aureus (300–500 mg/mL). WSME induced cell death in MDA-MB-231 cells and significantly inhibited their growth (IC₅₀: 66 μg/mL, P value<0.05) without affecting normal Vero cells in the studied range of 25–400 μg/mL (IC₅₀: 6.09 mg/mL, P value>0.05).

Conclusions

The present study lends support to further testing of WSME against other cancer cell lines and animal models of cancer. These preclinical studies would provide further validation to its prospective use as an adjunct in human breast cancer therapy.

Keywords: Withania somnifera ; withanolides; cytotoxicity; breast cancer; antioxidant

Corresponding author: Rumana Ahmad, Associate Professor, Department of Biochemistry, Era’s Lucknow Medical College and Hospital, Era University, Sarfarazganj, Hardoi Road, Lucknow, 226003, UP, India, E-mail: rahmad@erauniversity.in

Funding source: Era Educational Trust, Era's Lucknow Medical College and Hospital, Era University

Award Identifier / Grant number: ELMC/R_Cell/I-180/2021

Acknowledgments

The authors gratefully acknowledge Dr. Meraj Ahmad, Professor, Dept. of Sociology, University of Lucknow, Lucknow, India, for being the inspiration and motivation behind the present work in view of his exceptional work and contribution in the area of social sciences, social work, humanities and public health.

Ethical approval: Not applicable.
Informed consent: Not applicable.
Author contributions: AS drafted the manuscript whereas RA planned, conceptualized and supervised the study as well as co-drafted, edited and revised the manuscript. AS carried out the in vitro biological activity experiments. SS, SM and MAK provided technical support as well as critically reviewed the manuscript.
Competing interests: Authors state no conflict of interest.
Research funding: Financial aid in the form of intramural research grant (vide ELMC/R_Cell/I-180/2021) from Era Educational Trust, Era’s Lucknow Medical College and Hospital, Era University, to RA and AS is gratefully acknowledged.

References

1. Siegel, RL, Giaquinto, AN, Jemal, A. Cancer statistics, 2024. Ca - Cancer J Clin 2024;74:12–49. https://doi.org/10.3322/caac.21820.Search in Google Scholar PubMed

2. Shiva, MP. Inventory of forestry resources for sustainable management and biodiversity conservation. New Delhi: Indus Publishing Company; 1996.Search in Google Scholar

3. Singh, N, Bhalla, M, de Jager, P, Gilca, M. An overview on Ashwagandha: a Rasayana (rejuvenator) of Ayurveda. Afr J Tradit, Complementary Altern Med 2011;8. https://doi.org/10.4314/ajtcam.v8i5s.9.Search in Google Scholar

4. Husain, I, Ahmad, R, Chandra, A, Raza, ST, Shukla, Y, Mahdi, F. Phytochemical characterization and biological activity evaluation of ethanolic extract of Cinnamomum zeylanicum. J Ethnopharmacol 2018;219:110–6. https://doi.org/10.1016/j.jep.2018.02.001.Search in Google Scholar PubMed

5. Srivastava, AN, Ahmad, R, Khan, MA. Evaluation and comparison of the in vitro cytotoxic activity of Withania somnifera methanolic and ethanolic extracts against MDA-MB-231 and vero cell lines. Sci Pharm 2015;84:41–59. https://doi.org/10.3797/scipharm.1507-13.Search in Google Scholar PubMed PubMed Central

6. Singleton, VL, Orthofor, R, Raventos, RML. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin–Ciocaltau reagent. Methods Enzymol 1999;299:152–78.10.1016/S0076-6879(99)99017-1Search in Google Scholar

7. Zhishen, J, Mengcheng, T, Jianming, W. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem 1999;64:555–9. https://doi.org/10.1016/s0308-8146(98)00102-2.Search in Google Scholar

8. Ali, BM, Boothapandi, M, Nasar, AS. Nitric oxide, DPPH and hydrogen peroxide radical scavenging activity of TEMPO terminated polyurethane dendrimers: data supporting antioxidant activity of radical dendrimers. Data Brief 2020;28:104972. https://doi.org/10.1016/j.dib.2019.104972.Search in Google Scholar PubMed PubMed Central

9. Malik, F, Singh, J, Khajuria, A, Suri, KA, Satti, NK, Singh, S, et al.. A standardized root extract of Withania somnifera and its major constituent withanolide-A elicit humoral and cell-mediated immune responses by up regulation of Th1-dominant polarization in BALB/c mice. Life Sci 2007;80:1525–38. https://doi.org/10.1016/j.lfs.2007.01.029.Search in Google Scholar PubMed

10. Hahm, ER, Moura, MB, Kelley, EE, Van Houten, B, Shiva, S, Singh, SV. Withaferin A-induced apoptosis in human breast cancer cells is mediated by reactive oxygen species. PLoS One 2011;6:e23354. https://doi.org/10.1371/journal.pone.0023354.Search in Google Scholar PubMed PubMed Central

11. Sandipan Chatterjee, SC, Shatakshi Srivastava, SS, Asna Khalid, AK, Niharika Singh, NS, Sangwan, RS, Sidhu, OP, et al.. Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts. Phytochemistry 2010;71:1085–94. https://doi.org/10.1016/j.phytochem.2010.04.001.Search in Google Scholar PubMed

12. Yadav, B, Bajaj, A, Saxena, AK. In vitro anticancer activity of the root, stem and leaves of Withania somnifera against various human cancer cell lines. Indian J Pharmaceut Sci 2010;72:659–63. https://doi.org/10.4103/0250-474x.78543.Search in Google Scholar

13. Al-Hindawi, MK, Al-Khafaji, SH, Abdul-Nabi, MH. Anti-granuloma activity of Iraqi Withania somnifera. J Ethnopharmacol 1992;37:113–16. https://doi.org/10.1016/0378-8741(92)90069-4.Search in Google Scholar PubMed

14. Dorman, H, Bachmayer, O, Kosar, M, Hiltunen, R. Antioxidant properties of aqueous extracts from selected Lamiaceae species grown in Turkey. J Agric Food Chem 2004;52:762–70. https://doi.org/10.1021/jf034908v.Search in Google Scholar PubMed

15. Surveswaran, S, Cai, Y, Corke, H, Sun, M. Systematic evaluation of natural phenolic antioxidant from 133 Indian medicinal plants. Food Chem 2007;102:938–53. https://doi.org/10.1016/j.foodchem.2006.06.033.Search in Google Scholar

16. Piluzza, G, Bullitta, S. Correlations between phenolic content and antioxidant properties in twenty-four plant species of traditional ethnoveterinary use in the Mediterranean area. Pharmaceut Biol 2011;49:240–7. https://doi.org/10.3109/13880209.2010.501083.Search in Google Scholar PubMed

17. Paul, RK. In vitro antioxidant activity of Withania somnifera root. Int J Adv Res Comput Sci 2016;3:45–56.10.20431/2349-0403.0303006Search in Google Scholar

Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/jcim-2024-0185).

Received: 2024-05-24

Accepted: 2024-07-06

Published Online: 2024-08-20

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Supplementary Material

Articles in the same Issue

https://doi.org/10.1515/jcim-2024-0185

Keywords for this article

Withania somnifera; withanolides; cytotoxicity; breast cancer; antioxidant