ORIGINAL RESEARCH ARTICLE

Assessment of antibacterial, anti-inflammatory, and anti-cancer activities of Melia azedarach L. leaf extract

Mahendra Chikkamadaiah1 Dwaraknath Venkatesha2 Abhirami Dilkalal1 Ravindra Kembalu Narayana3 Venkatesh Hosur Narayanappa4 Satish Anandan5*
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1 Department of Studies in Botany, Bengaluru City University, Central College Campus, Bangalore, Karnataka, India
2 Department of Studies in Environmental Science, Tumkur University, Tumkur, Karnataka, India
3 Department of Studies in Botany, Karnataka State Open University, Mysuru, Karnataka, India
4 Research and Development, Miklens Bio-Private Limited, Bengaluru, Karnataka, India
5 Department of Clinical Nutrition and Dietetics, Sri Devaraj Urs Academy of Higher Education and Research, Tamaka, Kolar, India
CP 2024, 6(1), 2763
Submitted: 17 January 2024 | Accepted: 30 April 2024 | Published: 21 May 2024
© 2024 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

The purpose of the present study was to evaluate the antibacterial, anti-inflammatory, and cytotoxic properties of Melia azedarach leaf. The phytoconstituents found in the leaves were extracted with four different solvent systems based on their polarity index. Antimicrobial activity of selected plant extracts was evaluated using disc diffusion and micro-broth dilution methods against four different pathogenic bacterial strains. The ethanolic extract of M. azedarach leaf showed a significant inhibitory effect on the growth of Escherichia coli (15.07 mm) and Staphylococcus aureus (18.23 mm) (P < 0.005), followed by Bacillus subtilis and Salmonella typhi. Further, the mechanism of action was confirmed by live/dead cells analysis, which revealed the death of E. coli and S. aureus upon treatment with ethanolic extract. In vivo anti-inflammatory test conducted using carrageenan-induced rat paw edema model revealed that the 300 mg/kg ethanolic extract exhibited a significant anti-inflammatory activity of 51.78% compared with that of standard drug diclofenac (P < 0.001). Further, the cytotoxicity of ethanolic extract against human hepatocarcinoma cell lines (HepG2) was evaluated by MTT assay, and the findings showed a moderate level of toxicity against the HepG2 cell line with an IC50 value of 540.00 ± 0.6 μg/mL compared to doxorubicin. HepG2 cells treated with doxorubicin (2 μg/mL) and ethanolic extract showed a 2.33- and 1.35-fold increase in p53 gene expression, respectively. Apoptosis activity was measured in terms of DNA laddering, which indicated the late stage of apoptosis. The results showed that the extract-treated cell lines induced DNA fragmentation, which was found to be a potent anti-cancer mechanism in HepG2 cells. This study corroborated that the leaf extract of M. azedarach is a good source of active phytochemicals, with promising biological activities.

Keywords
Anti-inflammatory
Apoptosis
Carrageenan
HepG2
MTT assay
p53 gene
RT-PCR
Funding
The study is supported by the University Grant Commission, Major Research Project (No.: F.No.37- 460/2009[SR]).
References
  1. Pan SY, Litscher G, Gao SH, et al. Historical perspective of traditional indigenous medical practices: The current renaissance and conservation of herbal resources. Evid Based Complement Alternat Med. 2014;2014:525340. doi: 10.1155/2014/525340

 

  1. Pferschy-Wenzig EM, Bauer R. The relevance of pharmacognosy in pharmacological research on herbal medicinal products. Epilepsy Behav. 2015;52:344-362. doi: 10.1016/j.yebeh.2015.05.037

 

  1. Alamgir ANM. Therapeutic Use of Medicinal Plants and their Extracts: Volume 1. United States: Springer International Publishing AG; 2017. doi: 10.1007/978-3-319-92387-1

 

  1. Ferlay J, Colombet M, Soerjomataram I, et al. Cancer statistics for the year 2020: An overview. Int J Cancer. 2021;149(4):778-789. doi: 10.1002/ijc.33588

 

  1. Parveen B, Parveen A, Parveen R, Ahmad S, Ahmad M, Iqbal M. Challenges and opportunities for traditional herbal medicine today with special reference to its status in India. Ann Phytomed. 2020;9(2):97-112. doi: 10.21276/ap.2020.9.2.8

 

  1. Sen A, Batra A. Melia azedarach L.-A paradise tree. J Funct Environ Bot. 2011;1(1):59-69.

 

  1. Carpinella MC, Defagó MT, Valladares G, Palacios SM. Role of Melia azedarach L. (Meliaceae) for the control of insects and Acari: Present status and future prospects. Adv Phytomed. 2006;3:81-123. doi: 10.1016/S1572-557X(06)03005-4

 

  1. Angamuthu D, Purushothaman I, Kothandan S, Swaminathan R. Antiviral study on Punica granatum L., Momordica charantia L., Andrographis paniculata Nees, and Melia azedarach L., to human herpes virus-3. Eur J Integr Med. 2019;28:98-108. doi: 10.1016/j.eujim.2019.04.008

 

  1. Yadav R, Kashaw V, Dudhe R. Standardization and anti-inflammatory potential of Amoora rohituka and Melia azedarach belongs to family Meliaceae. J Adv Sci Res. 2018;9(2):19-25.

 

  1. Redfern J, Kinninmonth M, Burdass D, Verran J. Using soxhlet ethanol extraction to produce and test plant material (essential oils) for their antimicrobial properties. J Microbiol Biol Educ. 2014;15(1):45-46. doi: 10.1128/jmbe.v15i1.656

 

  1. Harborne AJ. Phytochemical Methods a Guide to Modern Techniques of Plant Analysis. Berlin: Springer Science and Business Media; 1998.

 

  1. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Disk Susceptibility Tests. Wayne, PA: Clinical and Laboratory Standard Institute; 2013.

 

  1. Holland TL, Woods CW, Joyce M. Antibacterial susceptibility testing in the clinical laboratory. Infect Dis Clin North Am. 2009;23(4):757-790. doi: 10.1016/j.idc.2009.06.001

 

  1. Li R, Dhankhar D, Chen J, Cesario TC, Rentzepis PM. Determination of live: Dead bacteria as a function of antibiotic treatment. J Microbiol Methods. 2018;154:73-78. doi: 10.1016/j.mimet.2018.10.010

 

  1. Ecobichon DJ. The Basis of Toxicity Testing. Boca Raton: CRC Press; 1997.

 

  1. Singh M, Kumar V, Singh I, Gauttam V, Kalia AN. Anti-inflammatory activity of aqueous extract of Mirabilis jalapa Linn. Leaves. Pharmacognosy Res. 2010;2(6):364-367. doi: 10.4103/0974-8490.75456

 

  1. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1-2):55-63. doi: 10.1016/0022-1759(83)90303-4

 

  1. Vandghanooni S, Forouharmehr A, Eskandani M, et al. Cytotoxicity and DNA fragmentation properties of butylated hydroxyanisole. DNA Cell Biol. 2013;32(3):98-103. doi: 10.1089/dna.2012.1946

 

  1. Gaggia F, Capece A, Marino G. Antimicrobial and antifungal activity of different extracts from Melia azedarach L. On phytopathogenic bacteria and fungi of agro-food interest. In: Book of Abstracts of the First Symphosium in Horticulturae in Europe, 17-20. Vienna; 2008. p. 41.

 

  1. De CC Pinto N, Campos LM, Evangelista ACS, et al. Antimicrobial Annona muricata L. (Soursop) extract targets the cell membranes of Gram-positive and Gram-negative bacteria. Ind Crops Prod. 2017;107:332-340. doi: 10.1016/j.indcrop.2017.05.054

 

  1. Joux F, Lebaron P. Use of fluorescent probes to assess physiological functions of bacteria at single-cell level. Microbes Infect. 2000;2(12):1523-1535. doi: 10.1016/S1286-4579(00)01307-1

 

  1. Patil KR, Mahajan UB, Unger BS, et al. Animal models of inflammation for screening of anti-inflammatory drugs: Implications for the discovery and development of phytopharmaceuticals. Int J Mol Sci. 2019;20(18):4367. doi: 10.3390/ijms20184367

 

  1. Seibert K, Zhang Y, Leahy K, et al. Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Natl Acad Sci U S A. 1994;91(25):12013-12017. doi: 10.1073/pnas.91.25.1201

 

  1. Akacha M, Lahbib K, Remadi MD, Boughanmi NG. Antibacterial, antifungal and anti-inflammatory activities of Melia azedarach (L.) Ethanolic leaves extract. Bangladesh J Pharmacol. 2016;11(3):666-674. doi: 10.3329/bjp.v11i3.27000

 

  1. Zeng J, Ma RJ, Wang L, et al. Chemical constituents from the leaves of Melia azedarach. Nat Prod Res. 2019;33(19): 2860-2863. doi: 10.1080/14786419.2018.1501690

 

  1. Zhang L, Ismail MM, Rocchetti G, Fayek NM, Lucini L, Saber FR. The untargeted phytochemical profile of three Meliaceae species related to in vitro cytotoxicity and anti-virulence activity against MRSA Isolates. Molecules. 2022;27(2):435. doi: 10.3390/molecules27020435

 

  1. Elmore S. Apoptosis: A review of programmed cell death. Toxicol Pathol. 2007;35(4):495-516. doi: 10.1080/01926230701320337

 

  1. Youle RJ, Strasser A. The BCL-2 protein family: Opposing activities that mediate cell death. Nat Rev Mol Cell Biol. 2008;9(1):47-59. doi: 10.1038/nrm2308

 

  1. Joray MB, Villafañez F, González ML, et al. P53 tumor suppressor is required for efficient execution of the death program following treatment with a cytotoxic limonoid obtained from Melia azedarach. Food Chem Toxicol. 2017;109:888-897. doi: 10.1016/j.fct.2017.04.039

 

  1. Momtazi-Borojeni AA, Behbahani M, Sadeghi-Aliabadi H. Antiproliferative activity and apoptosis induction of crude extract and fractions of Avicennia marina. Iran J Basic Med Sci. 2013;16(11):1203-1208.
Conflict of interest
The authors declare that they have no competing interests.
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