KARAGENIN SEBAGAI MODEL INFLAMASI PADA KULIT MENCIT: GAMBARAN MAKROSKOPIK DAN HISTOPATOLOGIS

Sitarina Widyarini; Sugiyono Sugiyono; Yuli Purwandari Kristianingrum; Bambang Sutrisno

doi:10.22146/jsv.62689

KARAGENIN SEBAGAI MODEL INFLAMASI PADA KULIT MENCIT: GAMBARAN MAKROSKOPIK DAN HISTOPATOLOGIS

https://doi.org/10.22146/jsv.62689

Sitarina Widyarini^(1*), Sugiyono Sugiyono⁽²⁾, Yuli Purwandari Kristianingrum⁽³⁾, Bambang Sutrisno⁽⁴⁾

(1) Departemen Patologi Fakultas Kedokteran Hewan Universitas Gadjah Mada
(2) Departemen Patologi Fakultas Kedokteran Hewan Universitas Gadjah Mada
(3) Departemen Patologi Fakultas Kedokteran Hewan Universitas Gadjah Mada
(4) Departemen Patologi Fakultas Kedokteran Hewan Universitas Gadjah Mada
(*) Corresponding Author

Abstract

Penelitian ini bertujuan untuk membuat model inflamasi kulit pada punggung mencit betina galur Swiss dengan menggunakan karagenin dengan berbagai konsentrasi. Pada penelitian ini akan digunakan 45 ekor mencit betina galur Swiss umur 2 bulan yang dibagi dalam tiga kelompok secara acak. Kelompok I diberi karagenin 1%. Kelompok II diberi karagenin 2%, dan kelompok III diberi karagenin 4%. Induksi inflamasi dengan karagenin dilakukan melalui injeksi sub kutan karagenin dengan volume 0,1 mL pada kulit punggung mencit. Sebelum dilakukan injeksi karagenin tebal lipat kulit (skin-fold thickness) diukur dengan menggunakan jangka sorong digital. Selanjutnya setiap jam setelah injeksi sampai dengan jam ke- 6 pasca injeksi. Hewan coba selanjutnya dietanasi dengan cara dislokasi servikalis pada akhir jam ke-6. Organ kulit diambil dengan memotong kulit pada area dimana injeksi karagenin dilakukan dan selanjutnya diawetkan dengan formalin 10% untuk pemeriksaan histopatologis dengan pewarnaan hematoxylin eosin. Analisis data tebal lipat kulit dianalisis dengan Two Ways Anova dan diikuti dengan post hoc test Tukey test. Hasil penelitian memperlihatkan ada perbedaan bermakna pada tebal lipat kulit antar kelompok perlakuan (p<0,05). Injeksi subkutaneuos karagenin 1%, 2% dan 4% meningkatkan tebal lipat kulit sebesar 35%, 50% dan 60% pada jam pertama pasca injeksi (p<0,05). Inflamasi mild dan moderate pada bagian dermis ditemukan pasca injeksi karagenin 1% dan 2%. Infamasi severe pada bagian dermis dan panikulus ditemukan pasca injeksi karagenin 4%. Karagenin dapat dipakai sebagai model inflamasi akut pada kulit punggung mencit.

Keywords

inflamasi; karagenin; skin-fold thickness

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References

Ben-Kheder, S., Mzid M., Bardaa, S., Moalla, D., Shanoun, Z., Rebai, T., (2016). In vivo evaluation of the anti-inflammatory effect of Pistacia lentiscus Fruit Oil and Its Effects on Oxidative Stress. Evidence-Based Complementary and Alternative Medicine. Vo. 2016. http://dx.doi.org/10.1155/2016/6108203 Chen,L., Deng,H., Cui,H., Fang,J., Zuo,Z., Deng, J., Li, Y., Wang,X., and Ling Zhao, L. (2018). Inflammatory responses and inflammation-associated diseases in organs Oncotarget. 23; 9(6): 7204–7218. Cohen, SM., Ito, N., (2002). A critical review of the toxicological effects of carrageenan and processed eucheuma seaweed on the gastrointestinal tract, Critical Review of Toxicology. 32 (5) 413–444. D’Orazio,J., Jarrett,S., Amaro-Ortiz, A., Scott, T. (2013). UV Radiation and the Skin. International Journal of Molecular Science. 14(6): 12222–12248. Published online 2013 Jun 7. doi: 10.3390/ijms140612222 Fehrenbacher,JC., Vasko,MR., Duarte, JB. (2012). Models of Inflammation: Carrageenan- or Complete Freund’s Adjuvant-Induced Edema and Hypersensitivity in the Rat. Current Protocol In Pharmacology. 5: Unit 5.4 doi: 10.1002/0471141755.ph0504s56 Ferrero-Miliani L, Nielsen O, Andersen P, Girardin S. (2007). Chronic inflammation: importance of NOD2 and NALP3 in interleukin-1β generation. Clinical Experimental Immunology. 147:227–235. Huang S-Y, Feng C-W, Hung H-C, Chakraborty C, Chen C-H, et al. (2014) A Novel Zebrafish Model to Provide Mechanistic Insights into the Inflammatory Events in Carrageenan-Induced Abdominal Edema. PLoS ONE 9(8): e104414.doi:10.1371/ Kim, HK., (2016). Garlic Supplementation Ameliorates UV-Induced Photoaging in Hairless Mice by Regulating Antioxidative Activity and MMPs Expression. Molecules. 21(1): 70-80. doi: 10.3390/molecules21010070 Kitsukawa Y, Saito H, Suzuki Y, Kasanuki J, Tamura Y, Yoshida S. (1992). Effect of ingestion of eicosapentaenoic acid ethyl ester on carrageenan induced colitis in guinea pigs. Gastroenterology ;102(6):1859–66. Maillet M, Bonfils S, Lister RE. (1970). Carrageenan: effects in animals. Lancet; 2(7669):414–425. Marcus,R., Watt, J., (1981). Danger of carrageenan in foods and slimming recipes, Lancet 1 (8215):338. Medzhitov R. (2010). Inflammation : new adventures of an old flame. Cell. 140:771–776. Nathan C, Ding A. (2010). Nonresolving inflammation. Cell. 2010;140:871–882. Necas, J., Bartosikova, L., (2013). Carrageenan: a review. Veterinarni Medicina, 58, (4): 187–205 Posadas I., Bucci M., Roviezzo F., Rossi A., Parente L., Sautebin L., Cirino G (2004). Carrageenan-induced mouse paw edema is biphasic, aged-weight dependent and displays diferential nitric oxide cyclooxygenase-2 expression. British Journal of Pharmacology 142: 331-338 Poulsen E. (1973). Short-term peroral toxicity of undegraded carrageenan in pigs. Food Cosmet Toxicol.;11(2):219–27. Ravikiran, G., Elumalai, A., Eswarairah, MC., Naresh, V., (2012). An Annual Review of Anti inflammatory Medicinal Plants. International Journal of Phytotherapy, 2(1)23-27. Reanmongkol, W., Noppapan, T,., Subhadhirasakul, S., (2009). Antinociceptive, antipyretic, anti-inflammatory activities of Putranjiva roxburghii Wal. Leaf extract in experimental animals. Journal of Natural Medicine, Vol.63(3):290-296 Sharratt M, Grasso P, Carpanini F, Gangolli SD. 1970. Carrageenan ulceration as a model for human ulcerative colitis. Lancet;2(7679):932. Sini JM, Yarro AH., Ayanwuli LO., Aiyelero OM., Mallum SM., Gamaniel KS., (2010). Antinociceptive and anti-inflammatory activity of the aqueous extract of the root bark of Combretumsericeum in rodents. African Journal of Biotehnology, 9: 8872-8876 Stankov, S. V., 2012, Definition of Inflammation, Causes of Inflammation and Possible Anti- inflammatory Strategies, The Open Inflammation Journal, 5(1), 1-9. Stein C, Millan MJ, Herz A. 1988. Unilateral inflammation of the hindpaw in rats as a model of prolonged noxious stimulation: alterations in behavior and nociceptive thresholds. Pharmacology Biochemistry Behaviour.; 31:445–51. Suralkar, AA., Sarda, PS., Ghaisas, MM., Thakare, V., Deshpande, AD. 2008. In Vivo Animal Models for Evaluation of Antiinflammatory Activity. Article Review 6(2), 1-9. Szekalska M., Sosnowska, K., Tomcrykowa, M., Winnicka, K., Kasacka, I., Tomczyk, M., (2020). In vivo anti-inflamatory and anti-allergic activities of cymaroside evaluated by using hydrogel formulations. Biomedicine and Pharmacotheraphy, 121:109681 Tobacman,JK., 2001. Review of harmful gastrointestinal effects of carrageenan in animal experiments, Environmental Health Perspective, 109 (10): 983–994. Umamageswari, A., Kudagi, B., (2015). Anti-inflammatory and analgesic properties of Ocinum sactum: a comparative study using animal models. International Journal of Basic and Clinical Pharmacology. Vol. 4(5):981-986 Vinegar, R., Truax, JF., Selph, JL. 1976. Quantitative Studies of The Pathway to Acute Carrageenan Inflammation. Federation Proceedings. 35(13), 2447-2456. Vysakh, A., Jayesh, K., Helen, LR., Jyothis, M., Latha, MS., (2020) Acute oral toxicity and anti-inflammatory evaluation of mrthanoloic extract of Rotua aquatica roots in Wistar rats. Journal of Ayurverda and Integrative Medicine. 11:45-52. Vogel, HG., 2002. Drug Discovery and Evaluation: Pharmacological Assays. Second Edition, Springer Vorlag Berlin Heidelberg, pp 726-769 Watt, J., Marcus, R., 1981. Harmful effects of carrageenan fed to animals, Cancer Detect. Prev. 4 (1–4) 129–134. Widyarini S, Spinks N, Husband AJ, Reeve VE. 2001. Isoflavonoid compounds from red clover (Trifolium pratense) protect from inflammation and immune suppression induced by UV radiation. Photochem Photobiol.74(3):465-70 Widyarini, S., Domanski, D., Painter, N., Reeve, VE., (2006). Estrogen receptor signaling protects against immune suppression by UV radiation exposureProceeding National Academic Science U S A. 103(34): 12837–12842. Winter CA, Risley EA, Nuss GW. 1962. Carrageenin-induced edema in hind paw of the rat as an assay for antiiflammatory drugs. Proc Soc Exp Biol Med.; 111:544–7 Wei W., Feng, W., Xin,G., Niu Tingting , N., Zhanghe, Z., Haimina, C., Xiaojun, Y., 2016. Enhanced effect of κ-carrageenan on TNBS-induced inflammation in mice. Intl. Immunophamacol 39: 218–228 Wu, W., Wang, F., Gao, X., Niu, T., Zhu1, X., Yan, X., Chen, H., 2016. Synergistic effect of κ-carrageenan on oxazolone-induced inflammation in BALB/c mice. BMC Gastroenterology. 16:41 (DOI 10.1186/s12876-016-0459-7) Wu, W., Zhen, Z., Niu,T., Zhu, X., Gao, Y., Yan, J., Chen, Y.,Yan, X., Chen, H., 2017. 𝜅-Carrageenan Enhances Lipopolysaccharide-Induced Interleukin-8 Secretion by Stimulating the Bcl10-NF-𝜅B Pathway in HT-29 Cells and Aggravates C. freundii-Induced Inflammation in Mice. Mediators of Inflammation. http://dx.doi.org/10.1155/2017/8634865. Zendzian, RP., 2000. Dermal Absoption of Pesticidies in the Rat. Am Industrial Hygiene Assoc J. 61(4), 437-483. Zhou Y, Hong Y, Huang H. 2016. Triptolide Attenuates Inflammatory Response in Membranous Glomerulo-Nephritis Rat via Downregulation of NF-κB Signaling Pathway. Kidney and Blood Pressure Res. 41:901–910

DOI: https://doi.org/10.22146/jsv.62689