Therapeutic effect of Azadirachta indica (L.) flowers ethanolic extract (AIEE) against DMBA induced Breast cancer in Swiss Albino mice
Main Article Content
Abstract
Most often, aberrant cell proliferation and subsequent tumour formation are the causes of cancer development. Globally, millions of people are estimated to die from various types of cancer. Among various types of cancer, breast cancer is the most prevalent type in women throughout the world. It continues to have a high death rate even with different treatment approaches. Because of the complex natural etiology of breast cancer, its treatment is much challenging. Owing to substantial treatment hurdles, natural chemicals are being investigated as possible substitutes. The plant Azadirachta indica (L.); is extensively recognised since ancient times for various therapeutic uses in Ayurveda, Unani, and homeopathic medicine system. Scientifically, the phytoremedial compounds and anticancer properties of this plant have been widely studied in term of its preventive, protective, tumour-suppressive, immunomodulatory, and apoptotic actions against different cancers and their molecular processes. The present study focuses on therapeutic effect of Azadirachta indica (L.) flowers ethanolic extract (AIEE) against DMBA induced breast cancer in Swiss albino mice. Eighteen female mice weighing 30 ± 5 g were divided into two groups. Group-I: control (n=6) and Group-II: (n=12); were orally induced by DMBA (3 mg/ml dissolved in olive oil) at an interval of fourteen days until breast tumours (about 2.0 cm) were developed. Then after, Group-II was further divided into two sub-groups. Sub-Group-I; (n=6) as DMBA-Treated were sacrificed at same time and sub-group-II (n=6) were orally administered with AIEE (100mg/Kg body weight) for 5 weeks, and the last tumour volume were measured. The study results that, the mammary tumour volume significantly (p<0.05) reduced. Moreover, there was significant alteration (p<0.0001) observed at the level of hepatic and renal biomarkers. Conclusively, entire study depicts that AIEE administration possesses anti-proliferative activity by suppressing progression of breast tumour in mice model. AIEE administration also possesses significant (p<0.0001) improvement in the hepato-renal parameters. Therefore, Azadirachta indica flowers can be targeted as a novel and safe anti-cancer drug against breast cancer.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Abdel Moneim, A. E., Othman, M. S., & Aref, A. M. (2014). Azadirachta indica attenuates cisplatin-induced nephrotoxicity and oxidative stress. BioMed research international, 2014, 647131. https://doi.org/10.1155/2014/647131.
Abdel-Gaber, R., Hawsah, M. A., Al-Otaibi, T., Alojayri, G., Al-Shaebi, E. M., Mohammed, O. B., Elkhadragy, M. F., Al-Quraishy, S., & Dkhil, M. A. (2023). Biosynthesized selenium nanoparticles to rescue coccidiosis-mediated oxidative stress, apoptosis and inflammation in the jejunum of mice. Frontiers in immunology, 14, 1139899. https://doi.org/10.3389/fimmu.2023.1139899.
Akhouri, V., Kumari, M., & Kumar, A. (2020). Therapeutic effect of Aegle marmelos fruit extract against DMBA induced breast cancer in rats. Scientific reports, 10(1), 18016.
Ali, A. S., Nazar, M. E., Mustafa, R. M., Hussein, S., Qurbani, K., & Ahmed, S. K. (2024). Impact of heavy metals on breast cancer. World Academy of Sciences Journal, 6(1), 1-12.
Arnold, M., Morgan, E., Rumgay, H., Mafra, A., Singh, D., Laversanne, M., ... & Soerjomataram, I. (2022). Current and future burden of breast cancer: Global statistics for 2020 and 2040. The Breast, 66, 15-23.
Arora, N., Bansal, M. P., & Koul, A. (2013). Azadirachta indica acts as a pro-oxidant and modulates cell cycle associated proteins during DMBA/TPA induced skin carcinogenesis in mice. Cell biochemistry and function, 31(5), 385–394. https://doi.org/10.1002/cbf.2909.
Arumugam, A., Subramani, R., Nandy, S., Powell, S., Velazquez, M., Orozco, A., Galvez, A., & Lakshmanaswamy, R. (2016). Desacetyl nimbinene inhibits breast cancer growth and metastasis through reactive oxygen species mediated mechanisms. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 37(5), 6527–6537. https://doi.org/10.1007/s13277-015-4468-x.
Berthelot, M.P.E. (1859). Berthelot's Reaction Mechanism. Report de Chimie Applique, 2884.
Bhanwra, S., Singh, J., & Khosla, P. (2000). Effect of Azadirachta indica (Neem) leaf aqueous extract on paracetamol-induced liver damage in rats. Indian journal of physiology and pharmacology, 44(1), 64–68.
Bharati, S., Rishi, P., & Koul, A. (2014). Azadirachta indica modulates electrical properties and type of cell death in NDEA-induced hepatic tumors. Cell biochemistry and biophysics, 70(1), 383–390. https://doi.org/10.1007/s12013-014-9923-6.
Bones RW, Tausky HH (1945) Colorimetric determination of creatinine by the Jaffe reaction. J BiolChem 158:581–591.
Brown, A., Kumar, S., & Tchounwou, P. B. (2019). Cisplatin-based chemotherapy of human cancers. Journal of cancer science & therapy, 11(4).
Chaube, S. K., Shrivastav, T. G., Tiwari, M., Prasad, S., Tripathi, A., & Pandey, A. K. (2014). Neem (Azadirachta indica L.) leaf extract deteriorates oocyte quality by inducing ROS-mediated apoptosis in mammals. SpringerPlus, 3, 464. https://doi.org/10.1186/2193-1801-3-464.
Coffetti, G., Moraschi, M., Facchetti, G., & Rimoldi, I. (2023). The challenging treatment of cisplatin-resistant tumors: State of the art and future perspectives. Molecules, 28(8), 3407.
El-Beltagy, A. E. B. M., Saleh, A. M. B., Attaallah, A., & Gahnem, R. A. (2021). Therapeutic role of Azadirachta indica leaves ethanolic extract against diabetic nephropathy in rats neonatally induced by streptozotocin. Ultrastructural pathology, 45(6), 391–406. https://doi.org/10.1080/01913123.2021.1988015.
Elumalai, P., Ezhilarasan, D., & Raghunandhakumar, S. (2022). Molecular Targets of Nimbolide for Anti-Cancer Therapy: An Updated Review. Journal of environmental pathology, toxicology and oncology : official organ of the International Society for Environmental Toxicology and Cancer, 41(2), 69–88. https://doi.org/10.1615/JEnvironPatholToxicolOncol.2021040263.
Fawcett JK, Scott JE (1960) A rapid and precise method for the determination of urea. J ChemPathol 13:156–159.
Follain, G., Herrmann, D., Harlepp, S., Hyenne, V., Osmani, N., Warren, S. C., ... & Goetz, J. G. (2020). Fluids and their mechanics in tumour transit: shaping metastasis. Nature Reviews Cancer, 20(2), 107-124.
Fossati P, Prencipe L (1980). Enzymatic colorimetric method of the determination of uric acid in serum. ClinChem 26:227–231.
Gan, H., Zhang, Y., Zhou, Q., Zheng, L., Xie, X., Veeraraghavan, V. P., & Mohan, S. K. (2019). Zingerone induced caspase‐dependent apoptosis in MCF‐7 cells and prevents 7, 12‐dimethylbenz (a) anthracene‐induced mammary carcinogenesis in experimental rats. Journal of biochemical and molecular toxicology, 33(10), e22387.
Hamza, A. A., Khasawneh, M. A., Elwy, H. M., Hassanin, S. O., Elhabal, S. F., & Fawzi, N. M. (2022). Salvadora persica attenuates DMBA-induced mammary cancer through downregulation oxidative stress, estrogen receptor expression and proliferation and augmenting apoptosis. Biomedicine & Pharmacotherapy, 147, 112666.
Haque, E., Mandal, I., Pal, S., & Baral, R. (2006). Prophylactic dose of neem (Azadirachta indica) leaf preparation restricting murine tumor growth is nontoxic, hematostimulatory and immunostimulatory. Immunopharmacology and immunotoxicology, 28(1), 33–50. https://doi.org/10.1080/08923970600623632.
Hsieh, Y. H., Lee, C. H., Chen, H. Y., Hsieh, S. C., Lin, C. L., & Tsai, J. P. (2015). Induction of cell cycle arrest, DNA damage, and apoptosis by nimbolide in human renal cell carcinoma cells. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 36(10), 7539–7547. https://doi.org/10.1007/s13277-015-3477-0.
Hua, H., Zhang, H., Kong, Q., & Jiang, Y. (2018). Mechanisms for estrogen receptor expression in human cancer. Experimental hematology & oncology, 7(1), 1-11.
Huang, R., Chen, H., Liang, J., Li, Y., Yang, J., Luo, C., ... & Xie, X. (2021). Dual role of reactive oxygen species and their application in cancer therapy. Journal of Cancer, 12(18), 5543.
Hussain, S., Naseer, S., Waqas, M., Ashraf, S., Saddiqa, A., Ashraf, Y., ... & Javed, K. (2020). Factors Affecting the Rate of Breast Cancer; Role of Heavy Metals. International Journal of Economic and Environmental Geology, 11(3), 57-64.
Kandhare, A. D., Mukherjee, A. A., & Bodhankar, S. L. (2017). Neuroprotective effect of Azadirachta indica standardized extract in partial sciatic nerve injury in rats: Evidence from anti-inflammatory, antioxidant and anti-apoptotic studies. EXCLI journal, 16, 546–565. https://doi.org/10.17179/excli2017-161.
Kashyap, A. K., & Dubey, S. K. (2022). Molecular mechanisms in cancer development. In Understanding Cancer (pp. 79-90). Academic Press.
Kim, W. H., Song, H. O., Jin, C. M., Hur, J. M., Lee, H. S., Jin, H. Y., Kim, S. Y., & Park, H. (2012). The Methanol Extract of Azadirachta indica A. Juss Leaf Protects Mice Against Lethal Endotoxemia and Sepsis. Biomolecules & therapeutics, 20(1), 96–103. https://doi.org/10.4062/biomolther.2012.20.1.096.
Kind, P., & King, E. (1954). Estimation of plasma phosphatase by determination of hydrolysed phenol with amino-antipyrine. Journal of clinical Pathology, 7(4), 322.
Koul, A., Mohan, V., & Bharati, S. (2014). Azadirachta indica mitigates DMBA-induced hepatotoxicity: a biochemical and radiometric study. Indian journal of biochemistry & biophysics, 51(1), 37–45.
Lee, J. W., Ryu, H. W., Park, S. Y., Park, H. A., Kwon, O. K., Yuk, H. J., Shrestha, K. K., Park, M., Kim, J. H., Lee, S., Oh, S. R., & Ahn, K. S. (2017). Protective effects of neem (Azadirachta indica A. Juss.) leaf extract against cigarette smoke- and lipopolysaccharide-induced pulmonary inflammation. International journal of molecular medicine, 40(6), 1932–1940. https://doi.org/10.3892/ijmm.2017.3178.
Lei, S., Zheng, R., Zhang, S., Wang, S., Chen, R., Sun, K., ... & Wei, W. (2021). Global patterns of breast cancer incidence and mortality: A population‐based cancer registry data analysis from 2000 to 2020. Cancer Communications, 41(11), 1183-1194.
Martin, L., Zhang, Y., First, O., Mustieles, V., Dodson, R., Rosa, G., ... & Messerlian, C. (2022). Lifestyle interventions to reduce endocrine-disrupting phthalate and phenol exposures among reproductive age men and women: A review and future steps. Environment international, 107576.
Mitra, S., & Dash, R. (2018). Natural products for the management and prevention of breast cancer. Evidence-based complementary and alternative medicine, 2018.
Moga, M. A., Bălan, A., Anastasiu, C. V., Dimienescu, O. G., Neculoiu, C. D., & Gavriș, C. (2018). An overview on the anticancer activity of Azadirachta indica (Neem) in gynecological cancers. International journal of molecular sciences, 19(12), 3898.
Mukherjee, N., Saini, P., Mukherjee, S., Roy, P., & Sinha Babu, S. P. (2014). In vitro antifilarial activity of Azadirachta indica aqueous extract through reactive oxygen species enhancement. Asian Pacific journal of tropical medicine, 7(11), 841–848. https://doi.org/10.1016/S1995-7645(14)60147-4.
Omóbòwálé, T. O., Oyagbemi, A. A., Alaba, B. A., Ola-Davies, O. E., Adejumobi, O. A., Asenuga, E. R., Ajibade, T. O., Adedapo, A. A., & Yakubu, M. A. (2018). Ameliorative effect of Azadirachta indica on sodium fluoride-induced hypertension through improvement of antioxidant defence system and upregulation of extracellular signal regulated kinase 1/2 signaling. Journal of basic and clinical physiology and pharmacology, 29(2), 155–164. https://doi.org/10.1515/jbcpp-2017-0029.
Pooladanda, V., Thatikonda, S., Bale, S., Pattnaik, B., Sigalapalli, D. K., Bathini, N. B., Singh, S. B., & Godugu, C. (2019). Nimbolide protects against endotoxin-induced acute respiratory distress syndrome by inhibiting TNF-α mediated NF-κB and HDAC-3 nuclear translocation. Cell death & disease, 10(2), 81. https://doi.org/10.1038/s41419-018-1247-9.
Rahman, M. F., & Siddiqui, M. K. (2004). Biochemical effects of vepacide (from Azadirachta indica) on Wistar rats during subchronic exposure. Ecotoxicology and environmental safety, 59(3), 332–339. https://doi.org/10.1016/j.ecoenv.2003.07.013.
Rahman, M. F., Siddiqui, M. K., & Jamil, K. (2001). Effects of Vepacide (Azadirachta indica) on aspartate and alanine aminotransferase profiles in a subchronic study with rats. Human & experimental toxicology, 20(5), 243–249. https://doi.org/10.1191/096032701678227730.
Rajan, A., Nadhan, R., Latha, N. R., Krishnan, N., Warrier, A. V., & Srinivas, P. (2021). Deregulated estrogen receptor signaling and DNA damage response in breast tumorigenesis. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 1875(1), 188482.
Reddy, I. V., & Neelima, P. (2022). Neem (Azadirachta indica): A review on medicinal Kalpavriksha. International Journal of Economic Plants, 9(1), 59-63.
Reitman, S. and Frankel, S. (1957). Reitman and Frankel’s methods of estimating S.G.O.T. and S.G.P.T. Am. Journ. Clin. Path., 28: 56-63.
Ruslie, R. H., & Darmadi, D. (2020). Administration of neem (Azadirachta indica A. Juss) leaf extract decreases TNF-α and IL-6 expressions in dextran sodium sulfate-induced colitis in rats. Journal of advanced veterinary and animal research, 7(4), 744–749. https://doi.org/10.5455/javar.2020.g476.
Sahu, B., Sahu, A. K., Chennareddy, S. R., Soni, A., & Naithani, S. C. (2017). Insights on germinability and desiccation tolerance in developing neem seeds (Azadirachta indica): Role of AOS, antioxidative enzymes and dehydrin-like protein. Plant physiology and biochemistry : PPB, 112, 64–73. https://doi.org/10.1016/j.plaphy.2016.12.022.
Saini, A., Kumar, M., Bhatt, S., Saini, V., & Malik, A. (2020). Cancer causes and treatments. International Journal of Pharmaceutical Sciences and Research, 11(7), 3121-3134.
Sarkar, S., Singh, R. P., & Bhattacharya, G. (2021). Exploring the role of Azadirachta indica (neem) and its active compounds in the regulation of biological pathways: an update on molecular approach. 3 Biotech, 11(4), 178.
Seriana, I., Akmal, M., Darusman, D., Wahyuni, S., Khairan, K., & Sugito, S. (2021). Neem Leaf (Azadirachta indica A. Juss) Ethanolic Extract on the Liver and Kidney Function of Rats. The Scientific World Journal, 2021, 7970424. https://doi.org/10.1155/2021/7970424.
Singh, H., Kaur, M., Dhillon, J. S., Batra, M., & Khurana, J. (2017). Neem: a magical herb in endodontics. Stomatological Disease and Science, 1, 50-54.
Srivastava, M. K., & Raizada, R. B. (2007). Lack of toxic effect of technical azadirachtin during postnatal development of rats. Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association, 45(3), 465–471. https://doi.org/10.1016/j.fct.2006.09.010.
Tripathi, A., Shrivastav, T. G., & Chaube, S. K. (2012). Aqueous extract of Azadirachta indica (neem) leaf induces generation of reactive oxygen species and mitochondria-mediated apoptosis in rat oocytes. Journal of assisted reproduction and genetics, 29(1), 15–23. https://doi.org/10.1007/s10815-011-9671-0.
Turner, M. C., Andersen, Z. J., Baccarelli, A., Diver, W. R., Gapstur, S. M., Pope III, C. A., ... & Cohen, A. (2020). Outdoor air pollution and cancer: An overview of the current evidence and public health recommendations. CA: a cancer journal for clinicians, 70(6), 460-479.
Zheng, H., Shao, F., Martin, S., Xu, X., & Deng, C. X. (2017). WEE1 inhibition targets cell cycle checkpoints for triple negative breast cancers to overcome cisplatin resistance. Scientific reports, 7(1), 43517.