ANTI-INFLAMMATORY EFFECTS AND KERATINOCYTE REGENERATIVE POTENTIAL OF CASSIA ALATA (LINN) LEAF EXTRACTS AND THEIR IMPLICATIONS FOR WOUND HEALING

Abstract views: 81 / PDF downloads: 142

Authors

Keywords:

Cassia alata, anti-inflammatory, cytotoxicity, cell viability, proliferation, keratinocyte

Abstract

In this study, dried and ground plant leaves of Cassia alata were extracted with methanol and water, which were then subjected to various analyses. Morphological changes of human keratinocytes in response to plant extracts were observed with a differential interference contrast (DIC) microscope imaging system. The cell viability and proliferation effects of the plant extracts were also evaluated via colorimetric cytotoxicity assays and the CyQUANT® assay. Anti-inflammatory effects of the plant extracts were evaluated by flow cytometry with a cytometric bead array (CBA) kit and also cyclooxygenase (COX)-1, COX-2 and 5-lipoxygenase (5-LOX) enzyme inhibition assays. Results of the DIC microscopy assay indicated that Human keratinocytes treated with either the C. alata methanol (CAM) or aqueous (CAA) extracts (0.1-0.2 mg/mL) achieved 100% confluency. Cytotoxicity testing confirmed that the plant extracts were not toxic to keratinocyte cells at the concentrations used in the study. Only two cytokines i.e. interleukin (IL)-6 and IL-8 were detected by the CBA method, with the results confirming that none of the plant extracts induce a pro-inflammatory effect. Moreover, CAM and CAA extracts showed strong anti-inflammatory effects in suppressing IL-8. Both plant extracts (6.25-100 µg/mL) demonstrated strong inhibitory effects on COX-1 and 5-LOX enzymes compared to the standards i.e. indomethacin and nordihydroguaiaretic acid, respectively. However, inhibition of the COX-2 enzyme was less compared with indomethacin. The CAM extract at a concentration of 6.25 µg/mL, produced strong inhibition of COX-1 and 5-LOX enzymes, which was greater than that of the respective control treatments at the same concentration. Hence, our results indicate that the C. alata leaf extracts have a strong anti-inflammatory potential, which could be used to treat wounds and inflammation associated with the skin.

References

Hennebelle, T., Weniger, B., Joseph, H., Sahpaz, S. and Bailleul, F. (2009): Senna alata. Fitoterapia 80: 385–393.

Chatterjee, S., Chatterjee, S. and Dutta, S. (2012): An overview on the ethanophytopathological studies of Cassia alata - an important medicinal plant and the effet of VAM on its growth and productivity. International Journal of Research in Botany 2(4): 13-19.

Saito, S., Silva, G., Santos, R.X., Gosmann, G., Pungartnik, C. and Brendel, M. (2012): Astragalin from Cassia alata Induces DNA Adducts in Vitro and Repairable DNA Damage in the Yeast Saccharomyces cerevisiae. International Journal of Molecular Sciences 13: 2846-2862.

Okwu, D.E. and Nnmadi, F.U. (2011): Cannabinoid dronabinol alkaloid with antimicrobial activity from Cassia alata Linn. The Chemica Sinica 2(2): 247-254.

Meenupriya, J., Vinisha, A.S. and Priya, P. (2014): Cassia alata and Cassia auriculata - Review of their bioactive potential. World Journal of Pharmaceutical Science 2(12): 1760-1769.

Hemlata and Kalidhar, S.B. (1993): Alatinone, an anthraquinone from Cassia alata. Phytochemzstry 32(6): 1616-1617.

Wahab, A., Tahira, Begum, S., Ayub, A., Mahmood, I., Mahmood, T., Ahmad, A. and Fayyaz, N. (2014): Luteolin and kaempferol from cassia alata, antimicrobial and antioxidant activity of its methanolic extracts. FUUAST Journal of Biology 4(1): 1-5.

Saito, S.T., Trentin, D.d.S., JoseMacedo, A., Pungartnik, C., Gosmann, G., Silveira, J.d.D., Guecheva, T.N., Henriques, J.A.P. and Brendel, M. (2012): Bioguided Fractionation Shows Cassia alata Extract to Inhibit Staphylococcus epidermidis and Pseudomonas aeruginosa Growth and Biofilm Formation. Evidence-Based Complementary and Alternative Medicine 2012: 1-14.

Wegwu, M.O., Ayalogu, E.O. and Sule, J.O. (2005): Antioxidant protective effects of Cassia alata in rats exposed to carbon tetrachloride. Journal of Applied Sciences and Environmental Management 9: 77-80.

Patil, S.B., Naikwade, N.S., Kondawar, M.S., Magdum, C.S. and Awale, V.B. (2009): Traditional uses of plants for wound healing in the Sangli district, Maharashtra. International Journal of PharmTech Research 1(3): 876-878.

Muniandy, K., Gothai, S., Tan, W.S., Kumar, S., Esa, N.M., Chandramohan, G., Al-Numair, K. and Arulselvan, P. (2018): In Vitro Wound Healing Potential of Stem Extract of Alternanthera sessilis. Evidence-Based Complementary and Alternative Medicine 2018: 1-14.

Wang, Z., Wang, Y., Farhangfar, F., Zimmer, M. and Zhang, Y. (2012): Enhanced Keratinocyte Proliferation and Migration in Co-culture with Fibroblasts. Plos One 7(7): 1-12.

Mcdougall, S., Dallon, J., Sherratt, J. and Maini, P. (2006): Fibroblast migration and collagen deposition during dermal wound healing: mathematical modelling and clinical implications. Philosophical Transactions of the Royal Society A 364: 1385–1405.

Wilson, V.G. (2014): Growth and Differentiation of HaCaT Keratinocytes. Methods in Molecular Biology 1195: 33–41.

Colombo, I. Sangiovanni, E. Maggio, R. Mattozzi, C. Zava, S. Corbett, Y. Fumagalli, M. Carlino, C. Corsetto, P.A. Scaccabarozzi, D. Calvieri, S. Gismondi, A. Taramelli, D. and DellAgli, M. (2017): HaCaT Cells as a Reliable In Vitro Differentiation Model to Dissect the Inflammatory/Repair Response of Human Keratinocytes. Mediators of Inflammation 2017: 1-13.

Ricciotti, E. and FitzGerald, G.A. (2011): Prostaglandins and Inflammation. Arteriosclerosis, Thrombosis and Vascular Biology 31(5): 986-1000.

Kapewangolo, P., Omolo, J.J., Bruwer, R., Fonteh, P. and Meyer, D. (2015): Antioxidant and anti-inflammatory activity of Ocimum labiatum extract and isolated labdane diterpenoid. Journal of Inflammation 12(1): 1-13.

Schreml, S., Szeimies, R.M., Prantl, L., Karrer, S., Landthaler, M. and Babilas, P. (2010): Oxygen in acute and chronic wound healing. British Journal of Dermatology 163(2): 257–268.

Barrientos, S., Stojadinovic, O., Golinko, M.S., Brem, H. and Tomic-Canic, M. (2008): Growth factors and cytokines in wound healing. Wound Repair and Regeneration 16: 585–601.

Broughton, G., Janis, J.E. and Attinger, C.E. (2006): The basic science of wound healing. Plastic and Reconstructive Surgery 117(7): 12S-34S.

Zhao, P.-W., Jiang, W.-G., Wang, L., Jiang, Z.-Y., Shan, Y.-X. and Jiang, Y.-F. (2014): Plasma Levels of IL-37 and Correlation with TNF-a, IL-17A, and Disease Activity during DMARD Treatment of Rheumatoid Arthritis. Plos One 9(5): 1-7.

Fujisawa, H., Wang, B., Sauder, D.N. and Kondo, S. (1997): Effects of Interferons on the Production of Interleukin-6 and Interleukin-8 in Human Keratinocytes. Journal of Interferon and Cytokine Research 17: 347-353.

Feng, L.V., Wei, Y., Yang, Y.U., Jun-bo, H.U., Bin, Z. and Jing, W. (2013): Effects of the 24 N-terminal Amino Acids of p55PIK on Endotoxinstimulated Release of Inflammatory Cytokines by HaCaT Cells. Journal of Huazhong University of Science and Technology - Medical Science 33(4): 587-593.

Behm, B., Babilas, P., Landthaler, M. and Schreml, S. (2012): Cytokines, chemokines and growth factors in wound healing. Journal of the European Academy of Dermatology and Venereology 26: 812–820.

Efron, P.A. and Moldawer, L.L. (2004): Cytokines and Wound Healing: The Role of Cytokine and Anticytokine Therapy in the Repair Response. Journal of Burn Care and Rehabilitation 25: 149-160.

Franz, A.R., Steinbach, G., Kron, M. and Pohlandt, F. (2001): Interleukin-8: a valuable tool to restrict antibiotic therapy in newborn infants. Acta Paediatrica 90: 1025-1032.

Attila, T., Jorg, H., Roy, C. and Rudi, V. (2003): Cytometric bead array to measure six cytokines in twenty five microliters of serum. Clinical Chemistry 49(6): 1000-1002.

Thangarasu, P., Manikandan, A. and Thamaraiselvi, S. (2019): Discovery, synthesis and molecular corroborations of medicinally important novel pyrazoles; drug efficacy determinations through in silico, in vitro and cytotoxicity validations. Bioorganic Chemistry 86: 410–419.

Atatreh, N., Youssef, A.M., Ghattas, M.A., Sorkhy, M.A., Alrawashdeh, S., Al-Harbi, K.B., El-Ashmawy, I.M., Almundarij, T.I., Abdelghani, A.A. and Abd-El-Aziz, A.S. (2019): Anti-inflammatory drug approach: Synthesis and biological evaluation of novel pyrazolo[3,4-d] pyrimidine compounds. Bioorganic Chemistry 86: 393–400.

Yao, C. and Narumiya, S. (2019): Prostaglandin-cytokine crosstalk in chronic inflammation. British Journal of Pharmacology 176: 337–354.

Elzahhar, P.A. Alaaeddine, R. Ibrahim, T.M. Nassra, R. Ismail, A. Chua, B.S.K. Frkic, R.L. Bruning, J.B. Wallner, N. Knape, T. Knethen, A.v. Labib, H. El-Yazbi, A.F. and Belal, A.S.F. (2019): Shooting three inflammatory targets with a single bullet: Novel multi-targeting anti-inflammatory glitazones. European Journal of Medicinal Chemistry 167: 562-582.

Adebayo, S.A. and Amoo, S.O. (2019): South African botanical resources: A gold mine of natural pro-inflammatory enzyme inhibitors? South African Journal of Botany 123: 214–227.

Ondua, M., Njoya, E.M., Abdalla, M.A. and McGaw, L.J. (2019): Anti-inflammatory and antioxidant properties of leaf extracts of eleven South African medicinal plants used traditionally to treat inflammation. Journal of Ethnopharmacology 234: 27–35.

Kicel, A., Owczarek, A., Gralak, P., Ciszewski, P. and Olszewska, M.A. (2019): Polyphenolic profile, antioxidant activity, and pro-inflammatory enzymes inhibition of leaves, flowers, bark and fruits of Cotoneaster integerrimus: A comparative study. Phytochemistry Letters 30: 349–355.

Phan, M.A.T., Bucknall, M.P. and Arcot, J. (2019): Interferences of anthocyanins with the uptake of lycopene in Caco-2 cells, and their interactive effects on anti-oxidation and anti-inflammation in vitro and ex vivo. Food Chemistry 276: 402–409.

Sikandan, A., Shinomiya, T. and Nagahara, Y. (2018): Ashwagandha root extract exerts antiinflammatory effects in HaCaT cells by inhibiting the MAPK/NFκB pathways and by regulating cytokines. International Journal of Molecular Medicine 42: 425-434.

Sigma-Aldrich, Cell Proliferation Kit I (MTT). 2020, Roche Diagnostics GmbH: Germany.

Hong, C.-E. and Lyu, S.-Y. (2011): Anti-inflammatory and Anti-oxidative Effects of Korean Red Ginseng Extract in Human Keratinocytes. Immune Network 11(1): 42-49.

BD-Bioscience, BD Cytometric Bead Array (CBA) Human Inflammatory Cytokines Kit Instruction Manual. 2015, BD Biosciences: Becton, Dickinson and Company.

Gierse, J.K. and Koboldt, C.M. (2001): Cyclooxygenase assays. Current protocols in pharmacology 3(1): 1-16.

Baylac, S. and Racine, P. (2003): Inhibition of 5-lipoxygenase by essential oils and other natural fragrant extracts. International Journal of Aromatherapy 13(2): 138-142.

Kamatou, G.P.P., Viljoen, A.M. and Steenkamp, P. (2010): Antioxidant, anti-inflammatory activities and HPLC analysis of South African Salvia species. Food Chemistry 119(2): 684-688.

Yalavarthi, C. and Thiruvengadarajan, V.S. (2013): A review on identification stratergy of phyto constituents present in herbal plants. International Journal of Research in Pharmaceutical Sciences 4(2): 123-140.

Moriyama, H., Iizuka, T., Nagai, M., Miyataka, H. and Satoh, T. (2003): Antiinflammatory Activity of Heat-treated Cassia alata Leaf Extract and Its Flavonoid Glycoside. Yakugaku Zasshi - Journal of the Pharmaceutical Society of Japan 123(7): 607―611.

Moriyama, H., Iizuka, T. and Nagai, M. (2001): A Stabilized Flavonoid Glycoside in Heat-Treated Cassia alata Leaves and Its Structural Elucidation. Yakugaku Zasshi - Journal of the Pharmaceutical Society of Japan 121(11): 817-820.

Babitha, S., Nguyen, D.H., Park, S.-J., Shin, J.-H., Reyes, G.A., Caburian, A. and Kim, E.-K. (2010): Potential of Cassia alata Leaf Extract in Inducing Differentiation and Migration of Mouse Melanoblasts. Biotechnology and Bioprocess Engineering 15: 1071-1076.

Niles, A.L., Moravec, R.A. and Riss, T.L. (2009): In Vitro Viability and Cytotoxicity Testing and Same-Well Multi-Parametric Combinations for High Throughput Screening. Current Chemical Genomics 3: 33-41.

Rezk, A., Al-Hashimi, A., John, W., Schepker, H., Ullrich, M.S. and Brix, K. (2015): Assessment of cytotoxicity exerted by leaf extracts from plants of the genus Rhododendron towards epidermal keratinocytes and intestine epithelial cells. BMC Complementary and Alternative Medicine 15: 1-18.

Mosmann, T. (1983): Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of lmmunological Methods 65: 55-63.

Jones, L.J., Gray, M., Yue, S.T., Haugland, R.P. and Singer, V.L. (2001): Sensitive determination of cell number using the CyQUANT cell proliferation assay. Journal of Immunological Methods 254: 85–98.

Sriwilaijaroen, N., Kelly, J.X., Riscoe, M. and Wilairat, P. (2004): CyQUANT cell proliferation assay as a fluorescence based Method for in vitro screening of antimalarial activity. Southeast Asian Journal of Tropical Medicine and Public Health 35(4): 840-844.

Levy, A. and Lewis, A. (2011): Cassia alata Leaf Extract Induces Cytotoxicity in A549 Lung Cancer Cells via a Mechanism that is Caspase 8 Dependent. West Indian Medical Journal 60(6): 608-614.

Levy, A.S. and Carley, S.-K. (2012): Cytotoxic Activity of Hexane Extracts of Psidium Guajava L (Myrtaceae) and Cassia Alata L (Caesalpineaceae) in Kasumi-1 and OV2008 Cancer Cell Lines. Tropical Journal of Pharmaceutical Research 11(2): 201-207.

Huai, J., Jöckel, L., Schrader, K. and Borner, C. (2010): Role of caspases and non-caspase proteases in cell death. F1000 Biology Reports 2(48): 1-6.

Pereira, W.O. and Amarante-Mendes, G.P. (2011): Apoptosis: A Programme of Cell Death or Cell Disposal? Scandinavian Journal of Immunology 73: 401–407.

Raji, P., Sreenidhi, J., Sugithra, M., Renugadevi, K. and Samrot, A.V. (2015): Phytochemical Screening and Bioactivity Study of cassia alata Leaves. Biosciences, Biotechnology Research Asia 12(2): 291-296.

Pieme, C.A., Penlap, V.N., Ngogang, J., Kuete, V., Catros, V. and Moulinoux, J.P. (2009): In vitro effects of extract of Senna alata (Ceasalpiniaceae) on the polyamines produced by Leukaemia cells (L1210). Pharmacognosy Magazine 4(17): 8-13.

Jin, S.E., Ha, H., Shin, H.-K. and Seo, C.-S. (2019): Anti-Allergic and Anti-Inflammatory Effects of Kuwanon G and Morusin on MC/9 Mast Cells and HaCaT Keratinocytes. Molecules 24(265): 1-14.

Magcwebeba, T., Riedel, S., Swanevelder, S., Bouic, P., Swart, P. and Gelderblom, W. (2012): Interleukin-1α Induction in Human Keratinocytes (HaCaT): An In Vitro Model for Chemoprevention in Skin. Journal of Skin Cancer 2012: 393681.

Bassino, E., Gasparri, F. and Munaron, L. (2018): Pleiotropic Effects of White Willow Bark and 1,2-Decanediol on Human Adult Keratinocytes. Skin Pharmacol Physiol 31: 10–18.

Zampetti, A., Mastrofrancesco, A., Florji, E., Maresca, V., Picardo, M., Amerio, P. and Feliciani, C. (2009): Pro-inflammatory cytokine production in HaCaT cells treated by eosin: Implications for the topical treatment of psoriasis. International Journal of Immunopathology and Pharmacology 22(4): 1067-1075.

Sundaramoorthy, S., Gunasekaran, S., Arunachalam, S. and Sathiavelu, M. (2016): A Phytopharmacological Review on Cassia Species. Journal of Pharmaceutical Sciences and Research 8(5): 260-264.

Villasenor, I.M., Canlas, A.P., Pascua, M.P.I., Sabando, M.N. and Soliven, L.A.P. (2002): Bioactivity Studies on Cassia alata Linn. Leaf Extracts. Phytotherapy Research 16: S93–S96.

Lewis, A. and Levy, A. (2011): Anti-inflammatory Activities of Cassia alata Leaf Extract in Complete Freund's Adjuvant Arthritis in Rats. West Indian Medical Journal 60(6): 615-621.

Villasen, I.M., Canlas, A.P., Pascua, M.P.I., Sabando, M.N. and Soliven, L.A.P. (2002): Bioactivity Studies on Cassia alata Linn. Leaf Extracts. Phytotherapy Research 16: S93–S96.

Sagnia, B., Fedeli, D., Casetti, R., Montesano, C., Falcioni, G. and Colizzi, V. (2014): Antioxidant and Anti-Inflammatory Activities of Extracts from Cassia alata, Eleusine indica, Eremomastax speciosa, Carica papaya and Polyscias fulva Medicinal Plants Collected in Cameroon. Plos One 9(8): 1-10.

Green, J.A., Stockton, R.A., Johnson, C. and Jacobson, B.S. (2004): 5-Lipoxygenase and cyclooxygenase regulate wound closure in NIH/3T3 fibroblast monolayers. The American Journal of Physiology-Cell Physiology 287: C373–C383.

Claria, J. and Romano, M. (2005): Pharmacological Intervention of Cyclooxygenase-2 and 5-Lipoxygenase Pathways. Impact on Inflammation and Cancer. Current Pharmaceutical Design 11: 3431-3447.

Sudina, G.F., Pushkareva, M.A., Shephard, P. and Klein, T. (2008): Cyclooxygenase (COX) and 5-lipoxygenase (5-LOX) selectivity of COX inhibitors. Prostaglandins, Leukotrienes and Essential Fatty Acids 78: 99–108.

Brooks, S.J. (2018): Targeting Cytokines in the 5-LOX Pro-Inflammatory Pathway for Treatment Resistant Anorexia Nervosa. Journal of Molecular and Genetic Medicine 12(4): 1-7.

Blain, H., Boileau, C., Lapicque, F., Nedelec, E., Loeuile, D., Guillaume, C., Guaucher, A., Jeandel, C., Netter, P. and Jouzeau, J.-Y. (2002): Limitations of the in vitro whole blood assay for predicting the COX selectivity of NSAIDSs in clinical use. Journal of Clinical Pharmacology 53: 255-265.

Martel-Pelletier, J., Lajeunesse, D., Reboul, P. and Pelletier, J.P. (2003): Therapeutic role of dual inhibitors of 5-LOX and COX, selective and non-selective non-steroidal anti-inflammatory drugs. Annals of the Rheumatic Diseases 62: 501-509.

Cipollone, F. and Patrono, C. (2002): Cyclooxygenase-2 Polymorphism: Putting a Brake on the Inflammatory Response to Vascular Injury? Arteriosclerosis, Thrombosis and Vascular Biology 22(10): 1516-1518.

Lin, H.-C., Lin, T.-H., Wu, M.-Y., Chiu, Y.-C., Tang, C.-H., Hour, M.-J., Liou, H.-C., Tu, H.-J., Yang, R.-S. and Fu, W.-M. (2014): 5-Lipoxygenase Inhibitors Attenuate TNF-a Induced Inflammation in Human Synovial Fibroblasts. Plos One 9(9): 1-11.

Simeone, A.-M., Nieves-Alicea, R., Mcmurtry, V.C., Colella, S., Krahe, R. and Tari, A.M. (2007): Cyclooxygenase-2 uses the protein kinase C/interleukin-8/urokinase-type plasminogen activator pathway to increase the invasiveness of breast cancer cells. International Journal of Oncology 30: 785-792.

Hinson, R.M., Williamst, J.A. and Shacter, E. (1996): Elevated interleukin 6 is induced by prostaglandin E2 in a murine model of inflammation: Possible role of cyclooxygenase-2. Proceedings of the National Academy of Sciences of the United States of America 93: 4885-4890.

Maihofner, C., Panayiotou, M., Charalambous, Bhambra, U., Lightfoot, T., Geisslinger, G. and Gooderham, N.J. (2003): Expression of cyclooxygenase-2 parallels expression of interleukin-1beta, interleukin-6 and NF-kappaB in human colorectal cancer. Carcinogenesis 24(4): 665-671.

Morita, I., Schindler, M., Regier, M.K., Otto, J.C., Hori, T., DeWitt, D.L. and Smith, W.L. (1995): Different inrecellular locations for prosterglandin endoperoxide H synthase-1 and -2. The Journal of Biological Chemistry 270(18): 10902-10908.

Brock, T.G., McNish, R.W., Bailie, M.B. and Peters-Golden, M. (1997): Rapid Import of Cytosolic 5-Lipoxygenase into the Nucleus of Neutrophils after in Vivo Recruitment and in Vitro Adherence. The Journal of Biological Chemistry 272(13): 8276–8280.

Downloads

Published

2022-09-28

How to Cite

Agampodi, V. A., & Collet, T. (2022). ANTI-INFLAMMATORY EFFECTS AND KERATINOCYTE REGENERATIVE POTENTIAL OF CASSIA ALATA (LINN) LEAF EXTRACTS AND THEIR IMPLICATIONS FOR WOUND HEALING. Journal of Applied Biological Sciences, 16(3), 503–526. Retrieved from https://jabsonline.org/index.php/jabs/article/view/1021

Issue

Section

Articles