Anti-Angiogenesis Screening of Moringa oleifera Pod Extracts by In-Ovo Chorioallantoic Membrane (CAM) Assay
Year 2023,
, 301 - 309, 01.12.2023
Nisha Shri Chengama Raju
,
Tee Siew Ying
Abstract
Moringa oleifera has many therapeutic benefits one out of it is anti-cancer property. Therefore, many researchers have been screening the therapeutic potential of Moringa oleifera. The main objective of this study was to screen and explore the angiogenesis inhibition potential of Moringa oleifera pod extracts. With the aim of screening anti-angiogenic potential, extracts of Moringa oleifera pods were prepared by decoction method. The extracts were subjected to preliminary phytochemical screening to identify the nature of phytochemicals present in the pods. In-ovo chorioallantoic membrane assay was chosen to achieve the objective of the study. Water-soluble extractive value (15.00% w/w) was higher than that of alcohol-soluble extractive value (3.89% w/w), indicating that the Moringa oleifera pods have more water-soluble constituents. Qualitative phytochemical screening revealed presence of flavonoids. Angiogenesis inhibition effect was studied and compared with sunitinib. Statistical analysis revealed highest anti-angiogenesis activity in 100% methanolic extract. Least effect was observed in 50% aqueous extract. Anti-angiogenic potential of 100% methanolic extract was statistically significant when compared with other study groups. It is concluded that Moringa oleifera pods exert anti-angiogenic potential and more intensified and diversified studies are needed to enable a thorough investigation of this plant components in anti-cancer treatment.
References
- 1. Ashraf MA. Phytochemicals as Potential Anticancer Drugs: Time to Ponder Nature’s Bounty. Biomed Res Int. 2020;2020:1–7.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013350/
- 2. Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2019. Cancer J Clin. 2019;69(1):7–34.
https://pubmed.ncbi.nlm.nih.gov/30620402/
- 3. Bhattacharya A, Tiwari P, Sahu PK, Kumar S. A Review of the Phytochemical and Pharmacological Characteristics of Moringa oleifera. J Pharm Bioallied Sci. 2018;10(4):181-91.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266645/
- 4. Ganguly S. Ayurveda and Pharmacy Finding Multivarious Application of Indigenous Herbs and Medicinal Plants for Traditional Therapy. Int J Ayu Pharm Chem. 2014;1(2):1-6.
https://www.researchgate.net/publication/268075751_Ayurveda_and_Pharmacy_finding_multivarious_application_of_indigenous_herbs_and_medicinal_plants_for_traditional_therapy
- 5. Paikra BK, Dhongade HKJ, Gidwani B. Phytochemistry and Pharmacology of Moringa oleifera Lam. J Pharmacopuncture. 2017;20(3):194–200.
https://doi.org/10.3831%2FKPI.2017.20.022
- 6. Al-Asmari AK, Albalawi SM, Athar MT, Khan AQ, Al-Shahrani H, Islam M. Moringa oleifera as an Anti-Cancer Agent against Breast and Colorectal Cancer Cell Lines. PLoS One. 2015;10(8):e0135814.
https://doi.org/10.1371/journal.pone.0135814
- 7. Saini RK, Sivanesan I, Keum YS. Phytochemicals of Moringa oleifera: A Review of Their Nutritional, Therapeutic and Industrial Significance. 3 Biotech. 2016;6(2):203-17.
https://doi.org/10.1007%2Fs13205-016-0526-3
- 8. Johnson IT. Phytochemicals and Cancer. Proc Nutr Soc. 2007:66(2):207–15.
https://doi.org/10.1017/s0029665107005459
- 9. Dandan K, Shaodan P, Jihua L, Yupo C. Study on the Effect of Extraction Process of Moringa Instant Tea on Its Sensory Quality. IOP Conf Ser: Earth Environ Sci. 2018;153(2):184-191.
https://iopscience.iop.org/article/10.1088/1755-1315/153/2/022026/pdf
- 10. Vongsak B, Sithisarn P, Mangmool S, Thongpraditchote S, Wongkrajang Y, Gritsanapan W. Maximizing Total Phenolics, Total Flavonoids Contents and Antioxidant Activity of Moringa oleifera Leaf Extract by the Appropriate Extraction Method. Indcrop. 2013;44:566–71.
http://dx.doi.org/10.1016%2Fj.indcrop.2012.09.021
- 11. Chaudhari RK, Girase NO. Determination of Soluble Extractives and Physico-Chemical Studies of Bark of Sesbania sesban (L) Merr. J Chem Pharm Res. 2015;7(8):657–60.
https://www.jocpr.com/articles/determination-of-soluble-extractives-and-physicochemical-studies-of-bark-of-sesbania-sesban-l-merr.pdf
- 12. Centers for Disease Control and Prevention. Sterilizing Practices - Guideline for Disinfection and Sterilization in Healthcare Facilities [document on the internet]. 2020. Available from: https://www.cdc.gov/infectioncontrol/guidelines/disinfection/sterilization/sterilizing-practices.html
- 13. Deryugina EI, Quigley JP. Chapter 2. Chick Embryo Chorioallantoic Membrane Models to Quantify Angiogenesis Induced by Inflammatory and Tumor Cells or Purified Effector Molecules. Methods Enzymol. 2008;444:21–41.
https://doi.org/10.1016/s0076-6879(08)02802-4
- 14. Kunz P, Schenker A, Sähr H, Lehner B, Fellenberg J. Optimization of the Chicken Chorioallantoic Membrane Assay as Reliable in Vivo Model for the Analysis of Osteosarcoma. PLoS One. 2019;14(4):e0215312.
https://doi.org/10.1371/journal.pone.0215312
- 15. Alexa, Anna. Incubating Part 2 - Setting and Candling. The Pioneer Chicks [document on the internet]. 2020. Available from: https://www.thepioneerchicks.com/incubating-setting-and-candling
- 16. Lokman NA, Elder ASF, Ricciardelli C, Oehler MK. Chick Chorioallantoic Membrane (CAM) Assay as an In Vivo Model to Study the Effect of Newly Identified Molecules on Ovarian Cancer Invasion and Metastasis. Int J Mol Sci. 2012;13(8):9959–70.
https://doi.org/10.3390/ijms13089959
- 17. Sonawane H, Shinde A, Jadhav J. Evaluation of Anti-Angiogenic Potential of Mentha arvensis Linn. Leaf Extracts Using Chorioallantoic Membrane Assay. World J Pharm Res. 2016;5(2): 677–89.
https://wjpr.net/abstract_file/4565
- 18. Gopalkrishnan B, Deb P, Roy C. Pharmacognostical Studies of Moringa oleifera Lam. Seeds. J Pharmacogn Phytochem. 2019;8(6):741–4.
https://www.phytojournal.com/archives/2019/vol8issue6/PartL/8-6-18-471.pdf
- 19. Siyanbola TO, Edobor-Osoh A, Ajanaku CO, Akinsiku AA, Adedapo EA, Aladesuyi O, et al. Nutritional and Physico-Chemical Evaluations of Moringa oleifera Seedlings and Oil. J Int Assoc Adv Technol Sci. 2015;1(1):1-5.
https://doi.org/10.53555/nnas.v2i7.677
- 20. Senguttuvan J, Paulsamy S, Karthika K. Phytochemical Analysis and Evaluation of Leaf and Root Parts of the Medicinal Herb, Hypochaeris radicata L. for in Vitro Antioxidant Activities. Asian Pac J Trop Biomed. 2014;4(Suppl 1):S359–S367.
https://doi.org/10.12980%2FAPJTB.4.2014C1030
- 21. Paulsamy S, Jeeshna MV. Preliminary Phytochemistry and Antimicrobial Studies of an Endangered Medicinal Herb Exacum bicolor Roxb. Res J Pharm. Biol Chem Sci. 2011;2(4):447–57.
https://hero.epa.gov/hero/index.cfm/reference/details/reference_id/8317408
- 22. Abdulkadir SI, Abdullahi NI, Sofowora A, Yahaya F, Ahmad AA, Hassan IA. Phytochemical Screening and Antimicrobial Activities of Ethanolic Extracts of Moringa oleifera Lam on Isolates of Some Pathogens. J App Pharm. 2015;7(4):203-10.
https://www.longdom.org/open-access/phytochemical-screening-and-antimicrobial-activities-of-ethanolic-extracts-ofmoringa-oleifera-lam-on-isolates-of-some-pathogens-1920-4159-1000203.pdf
- 23. Kadioglu O, Seo EJ, Efferth T. Targeting Angiogenesis by Phytochemicals. Med Aromat Plants. 2013;2(5):134-42.
https://www.longdom.org/open-access-pdfs/targeting-angiogenesis-by-phytochemicals-2167-0412.1000134.pdf
- 24. Kopustinskiene DM, Jakstas V, Savickas A, Bernatoniene J. Flavonoids as Anticancer Agents. Nutrients. 2020;12(2):457-81.
https://doi.org/10.3390/nu12020457
- 25. Martin NC, Pirie AA, Ford LV, Callaghan CL, McTurk K, Lucy D, et al. The Use of Phosphate Buffered Saline for the Recovery of Cells and Spermatozoa from Swabs. Sci Justice. 2006;46(3):179–84.
https://doi.org/10.1016/s1355-0306(06)71591-x
- 26. Mena AC, Pulido EG, Guillén-Ponce C. Understanding the Molecular-Based Mechanism of Action of the Tyrosine Kinase Inhibitor: Sunitinib. Anticancer Drugs. 2010;21(Suppl 1):S3-S11.
https://doi.org/10.1097/01.cad.0000361534.44052.c5
- 27. Gallego RP, Teves FG. Angiogenesis Modulatory Activity of SC-CO2 Leaf Extract of Guyabano (Annona muricata Linn.) Using Chick Embryo Chorioallantoic Membrane Assay. AAB Bioflux. 2019;7(3):206–16.
https://drive.google.com/file/d/1gsukNzp78iJgo6lIJOjdWnKTlpcgw_yI/view
- 28. Kei WW, Raju NSC. Anti-angiogenic screening of Moringa oleifera Leaves Extract Using Chorioallantoic Membrane Assay. Iraqi J Pharm Sci. 2022;31(1):225-232.
https://doi.org/10.31351/vol31iss1pp225-232
- 29. Certo G, Costa R, D’Angelo V, Russo M, Albergamo A, Dugo G, et al. Anti-Angiogenic Activity and Phytochemical Screening of Fruit Fractions from Vitex agnus Castus. Nat Prod Res. 2017;31(24),2850–6.
https://doi.org/10.1080/14786419.2017.1303696
- 30. Duraipandiyan V, Raja WRT, Al-Dhabi N, Savarimuthu I. Flavonoids-From Biosynthesis to Human Health: Chapter 13. Flavonoids: Anticancer Properties. 2017;287-303.
https://www.intechopen.com/chapters/56200
- 31. Salas GM, Totaan EV. Selected Philippine Herbal Plant Extracts as Angiogenesis Inhibitors Using Chick Chorioallantoic Membrane (CAM) Assay. Int Res J Biological Sci. 2015;4(9):28–32.
http://www.isca.in/IJBS/Archive/v4/i9/4.ISCA-IRJBS-2015-105.pdf
Anti-Angiogenesis Screening of Moringa oleifera Pod Extracts by In-Ovo Chorioallantoic Membrane (CAM) Assay
Year 2023,
, 301 - 309, 01.12.2023
Nisha Shri Chengama Raju
,
Tee Siew Ying
Abstract
Moringa oleifera has many therapeutic benefits one out of it is anti-cancer property. Therefore, many researchers have been screening the therapeutic potential of Moringa oleifera. The main objective of this study was to screen and explore the angiogenesis inhibition potential of Moringa oleifera pod extracts. With the aim of screening anti-angiogenic potential, extracts of Moringa oleifera pods were prepared by decoction method. The extracts were subjected to preliminary phytochemical screening to identify the nature of phytochemicals present in the pods. In-ovo chorioallantoic membrane assay was chosen to achieve the objective of the study. Water-soluble extractive value (15.00% w/w) was higher than that of alcohol-soluble extractive value (3.89% w/w), indicating that the Moringa oleifera pods have more water-soluble constituents. Qualitative phytochemical screening revealed presence of flavonoids. Angiogenesis inhibition effect was studied and compared with sunitinib. Statistical analysis revealed highest anti-angiogenesis activity in 100% methanolic extract. Least effect was observed in 50% aqueous extract. Anti-angiogenic potential of 100% methanolic extract was statistically significant when compared with other study groups. It is concluded that Moringa oleifera pods exert anti-angiogenic potential and more intensified and diversified studies are needed to enable a thorough investigation of this plant components in anti-cancer treatment.
References
- 1. Ashraf MA. Phytochemicals as Potential Anticancer Drugs: Time to Ponder Nature’s Bounty. Biomed Res Int. 2020;2020:1–7.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7013350/
- 2. Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2019. Cancer J Clin. 2019;69(1):7–34.
https://pubmed.ncbi.nlm.nih.gov/30620402/
- 3. Bhattacharya A, Tiwari P, Sahu PK, Kumar S. A Review of the Phytochemical and Pharmacological Characteristics of Moringa oleifera. J Pharm Bioallied Sci. 2018;10(4):181-91.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6266645/
- 4. Ganguly S. Ayurveda and Pharmacy Finding Multivarious Application of Indigenous Herbs and Medicinal Plants for Traditional Therapy. Int J Ayu Pharm Chem. 2014;1(2):1-6.
https://www.researchgate.net/publication/268075751_Ayurveda_and_Pharmacy_finding_multivarious_application_of_indigenous_herbs_and_medicinal_plants_for_traditional_therapy
- 5. Paikra BK, Dhongade HKJ, Gidwani B. Phytochemistry and Pharmacology of Moringa oleifera Lam. J Pharmacopuncture. 2017;20(3):194–200.
https://doi.org/10.3831%2FKPI.2017.20.022
- 6. Al-Asmari AK, Albalawi SM, Athar MT, Khan AQ, Al-Shahrani H, Islam M. Moringa oleifera as an Anti-Cancer Agent against Breast and Colorectal Cancer Cell Lines. PLoS One. 2015;10(8):e0135814.
https://doi.org/10.1371/journal.pone.0135814
- 7. Saini RK, Sivanesan I, Keum YS. Phytochemicals of Moringa oleifera: A Review of Their Nutritional, Therapeutic and Industrial Significance. 3 Biotech. 2016;6(2):203-17.
https://doi.org/10.1007%2Fs13205-016-0526-3
- 8. Johnson IT. Phytochemicals and Cancer. Proc Nutr Soc. 2007:66(2):207–15.
https://doi.org/10.1017/s0029665107005459
- 9. Dandan K, Shaodan P, Jihua L, Yupo C. Study on the Effect of Extraction Process of Moringa Instant Tea on Its Sensory Quality. IOP Conf Ser: Earth Environ Sci. 2018;153(2):184-191.
https://iopscience.iop.org/article/10.1088/1755-1315/153/2/022026/pdf
- 10. Vongsak B, Sithisarn P, Mangmool S, Thongpraditchote S, Wongkrajang Y, Gritsanapan W. Maximizing Total Phenolics, Total Flavonoids Contents and Antioxidant Activity of Moringa oleifera Leaf Extract by the Appropriate Extraction Method. Indcrop. 2013;44:566–71.
http://dx.doi.org/10.1016%2Fj.indcrop.2012.09.021
- 11. Chaudhari RK, Girase NO. Determination of Soluble Extractives and Physico-Chemical Studies of Bark of Sesbania sesban (L) Merr. J Chem Pharm Res. 2015;7(8):657–60.
https://www.jocpr.com/articles/determination-of-soluble-extractives-and-physicochemical-studies-of-bark-of-sesbania-sesban-l-merr.pdf
- 12. Centers for Disease Control and Prevention. Sterilizing Practices - Guideline for Disinfection and Sterilization in Healthcare Facilities [document on the internet]. 2020. Available from: https://www.cdc.gov/infectioncontrol/guidelines/disinfection/sterilization/sterilizing-practices.html
- 13. Deryugina EI, Quigley JP. Chapter 2. Chick Embryo Chorioallantoic Membrane Models to Quantify Angiogenesis Induced by Inflammatory and Tumor Cells or Purified Effector Molecules. Methods Enzymol. 2008;444:21–41.
https://doi.org/10.1016/s0076-6879(08)02802-4
- 14. Kunz P, Schenker A, Sähr H, Lehner B, Fellenberg J. Optimization of the Chicken Chorioallantoic Membrane Assay as Reliable in Vivo Model for the Analysis of Osteosarcoma. PLoS One. 2019;14(4):e0215312.
https://doi.org/10.1371/journal.pone.0215312
- 15. Alexa, Anna. Incubating Part 2 - Setting and Candling. The Pioneer Chicks [document on the internet]. 2020. Available from: https://www.thepioneerchicks.com/incubating-setting-and-candling
- 16. Lokman NA, Elder ASF, Ricciardelli C, Oehler MK. Chick Chorioallantoic Membrane (CAM) Assay as an In Vivo Model to Study the Effect of Newly Identified Molecules on Ovarian Cancer Invasion and Metastasis. Int J Mol Sci. 2012;13(8):9959–70.
https://doi.org/10.3390/ijms13089959
- 17. Sonawane H, Shinde A, Jadhav J. Evaluation of Anti-Angiogenic Potential of Mentha arvensis Linn. Leaf Extracts Using Chorioallantoic Membrane Assay. World J Pharm Res. 2016;5(2): 677–89.
https://wjpr.net/abstract_file/4565
- 18. Gopalkrishnan B, Deb P, Roy C. Pharmacognostical Studies of Moringa oleifera Lam. Seeds. J Pharmacogn Phytochem. 2019;8(6):741–4.
https://www.phytojournal.com/archives/2019/vol8issue6/PartL/8-6-18-471.pdf
- 19. Siyanbola TO, Edobor-Osoh A, Ajanaku CO, Akinsiku AA, Adedapo EA, Aladesuyi O, et al. Nutritional and Physico-Chemical Evaluations of Moringa oleifera Seedlings and Oil. J Int Assoc Adv Technol Sci. 2015;1(1):1-5.
https://doi.org/10.53555/nnas.v2i7.677
- 20. Senguttuvan J, Paulsamy S, Karthika K. Phytochemical Analysis and Evaluation of Leaf and Root Parts of the Medicinal Herb, Hypochaeris radicata L. for in Vitro Antioxidant Activities. Asian Pac J Trop Biomed. 2014;4(Suppl 1):S359–S367.
https://doi.org/10.12980%2FAPJTB.4.2014C1030
- 21. Paulsamy S, Jeeshna MV. Preliminary Phytochemistry and Antimicrobial Studies of an Endangered Medicinal Herb Exacum bicolor Roxb. Res J Pharm. Biol Chem Sci. 2011;2(4):447–57.
https://hero.epa.gov/hero/index.cfm/reference/details/reference_id/8317408
- 22. Abdulkadir SI, Abdullahi NI, Sofowora A, Yahaya F, Ahmad AA, Hassan IA. Phytochemical Screening and Antimicrobial Activities of Ethanolic Extracts of Moringa oleifera Lam on Isolates of Some Pathogens. J App Pharm. 2015;7(4):203-10.
https://www.longdom.org/open-access/phytochemical-screening-and-antimicrobial-activities-of-ethanolic-extracts-ofmoringa-oleifera-lam-on-isolates-of-some-pathogens-1920-4159-1000203.pdf
- 23. Kadioglu O, Seo EJ, Efferth T. Targeting Angiogenesis by Phytochemicals. Med Aromat Plants. 2013;2(5):134-42.
https://www.longdom.org/open-access-pdfs/targeting-angiogenesis-by-phytochemicals-2167-0412.1000134.pdf
- 24. Kopustinskiene DM, Jakstas V, Savickas A, Bernatoniene J. Flavonoids as Anticancer Agents. Nutrients. 2020;12(2):457-81.
https://doi.org/10.3390/nu12020457
- 25. Martin NC, Pirie AA, Ford LV, Callaghan CL, McTurk K, Lucy D, et al. The Use of Phosphate Buffered Saline for the Recovery of Cells and Spermatozoa from Swabs. Sci Justice. 2006;46(3):179–84.
https://doi.org/10.1016/s1355-0306(06)71591-x
- 26. Mena AC, Pulido EG, Guillén-Ponce C. Understanding the Molecular-Based Mechanism of Action of the Tyrosine Kinase Inhibitor: Sunitinib. Anticancer Drugs. 2010;21(Suppl 1):S3-S11.
https://doi.org/10.1097/01.cad.0000361534.44052.c5
- 27. Gallego RP, Teves FG. Angiogenesis Modulatory Activity of SC-CO2 Leaf Extract of Guyabano (Annona muricata Linn.) Using Chick Embryo Chorioallantoic Membrane Assay. AAB Bioflux. 2019;7(3):206–16.
https://drive.google.com/file/d/1gsukNzp78iJgo6lIJOjdWnKTlpcgw_yI/view
- 28. Kei WW, Raju NSC. Anti-angiogenic screening of Moringa oleifera Leaves Extract Using Chorioallantoic Membrane Assay. Iraqi J Pharm Sci. 2022;31(1):225-232.
https://doi.org/10.31351/vol31iss1pp225-232
- 29. Certo G, Costa R, D’Angelo V, Russo M, Albergamo A, Dugo G, et al. Anti-Angiogenic Activity and Phytochemical Screening of Fruit Fractions from Vitex agnus Castus. Nat Prod Res. 2017;31(24),2850–6.
https://doi.org/10.1080/14786419.2017.1303696
- 30. Duraipandiyan V, Raja WRT, Al-Dhabi N, Savarimuthu I. Flavonoids-From Biosynthesis to Human Health: Chapter 13. Flavonoids: Anticancer Properties. 2017;287-303.
https://www.intechopen.com/chapters/56200
- 31. Salas GM, Totaan EV. Selected Philippine Herbal Plant Extracts as Angiogenesis Inhibitors Using Chick Chorioallantoic Membrane (CAM) Assay. Int Res J Biological Sci. 2015;4(9):28–32.
http://www.isca.in/IJBS/Archive/v4/i9/4.ISCA-IRJBS-2015-105.pdf