Baicalein, H2 O2 Kaynaklı L929 Hücre Hasarında Koruyucu Etkiye Sahiptir

Year 2023, Volume: 76 Issue: 1, 11 - 16, 25.05.2023

Pınar Bayram

https://doi.org/10.4274/atfm.galenos.2023.58815

Abstract

Objectives: In this study, it was aimed to investigate the possible protective effects of baicalein in H2O2-induced L929 fibroblast cell model by determining the parameters of oxidative stress (SOD) and inflammation process (TNF-alpha, TGF-beta, IL-10).
Materials and Methods: Baicalein was applied at different doses (200, 100 and 80, μM) to L929 fibroblast cells. After 23 hours of incubation, 1 mM
H2O2 was added and cell viability was test by CVDK8. Then, fluorescent diacetate (FDA)-propidium iodide (PI) staining was performed to determine
the ratio of live and dead cells. The levels of TNF-alpha, IL10, TGF-beta and the activity of SOD were determined by ELISA.
Results: After 1 h incubation with 1 mM H2O2, the viability decreased by approximately 77%. It was observed that cell viability increased the most
in the B100 group, with/without H2O2. The cell viability increased after 24 h of baicalein and H2O2 co-incubation. The 100 μM baicalein application
reduced the damage of H2O2 up to 63%. When the levels of TNF-alpha were examined, a significant increase was observed in the H2O2-damage group
compared to control group. Additionally, the levels of IL-10 and TGF-beta increased in H2O2-damage group compared to control group; however, the
levels of those cytokines were decreased in baicalein application groups.
Conclusion: Pretreatment with baicalein (especially 200 μM and 100 μM) results in significant findings including the suppression of inflammatory
cytokines, TNF-alpha, TGF-beta, IL-10 and increase of SOD parameter, SOD, in L929 cells. Taking everything into account, baicalein which is a natural
product may be an alternative therapeutic option for prevention of cellular SOD and inflammatory processes. To better understand the potential
beneficial effects of baicalein, molecular mechanisms and cellular targets should be investigated.

Keywords

Baicalein , Hydrogen Peroxide , Inflammation , Oxidative Stress , L929

Ethical Statement

Ethics Committee Approval: This study is a cell culture study and therefore ethics committee approval was not obtained. Informed Consent: Cell culture study. Peer-reviewed: Externally peer-reviewed. Financial Disclosure: The author declared that she did not receive any financial support during this article

Supporting Institution

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Project Number

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Thanks

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Baicalein, H2O2 Kaynaklı L929 Hücre Hasarında Koruyucu Etkiye Sahiptir

Abstract

Amaç: Mevcut çalışmada, Baicalein’in H2O2 ile indüklenmiş L929 fibroblast hücre hasarı modelinde oksidatif stres (SOD) ve enflamasyon süreci (TNFalfa, TGF-beta, IL-10) parametreleri üzerine olan koruyucu etkilerinin araştırılması amaçlanmıştır.
Gereç ve Yöntem: L929 fibroblast hücrelerine farklı dozlarda (200, 100 ve 80 μM) Baicalein uygulandı. Yirmi üç saatlik inkübasyonun ardından 1 mM H2O2 eklendi ve hücre canlılığı CVDK8 ile test edildi. Daha sonra canlı ve ölü hücrelerin oranını belirlemek için floresan diasetat (FDA)-propidyum iyodür (PI) boyaması yapıldı. TNF-alfa, IL10, TGF-beta seviyeleri ve SOD aktivitesi ELISA ile belirlendi.

Bulgular: 1 mM H2O2 ile 1 saatlik inkübasyondan sonra hücre canlılığı yaklaşık %77 oranında azaldı. Hasarsız ya da H2O2 hasarlı gruplarda, hücre canlılığında artışın en fazla B100 grubunda olduğu görüldü. Baicalein ve H2O2’nin birlikte inkübasyonundan 24 saat sonra hücre canlılığının arttığı tespit edildi. 100 μM Baicalein uygulaması, H2O2’nin hücreleri öldürme oranını %63’e kadar azalttı. TNF-alfa düzeyleri incelendiğinde H2O2-hasarlı grupta kontrol grubuna göre anlamlı artış gözlendi. Ek olarak, IL-10 ve TGF-beta seviyeleri H2O2-hasarlı grupta kontrol grubuna göre arttı; ancak bu
sitokinlerin seviyeleri Baicalein uygulama gruplarında azaldı.

Sonuç: Baicalein (özellikle 200 μM ve 100μM) uygulaması sonucunda L929 hücrelerinde enflamatuvar sitokinler olan TNF-alfa, TGF-beta, IL- 10’un baskılanması ve SOD parametresi olan SOD’nin artması gibi önemli bulgular elde edilmiştir. Tüm veriler ışığında, doğal bir etken madde olan baicalein, hücresel SOD ve enflamatuvar süreçlerin önlenmesi için alternatif bir tedavi seçeneği potansiyeline sahiptir. Baicalein’in potansiyel yararlı etkilerini daha iyi anlamak için moleküler mekanizmalar ve hücresel hedeflerin araştırılmasına yönelik yeni çalışmalara ihtiyaç vardır.

Keywords

Baicalein , Hidrojen Peroksit , Enflamasyon , Oksidatif Stres , L929

Ethical Statement

Ethics Committee Approval: This study is a cell culture study and therefore ethics committee approval was not obtained. Informed Consent: Cell culture study. Peer-reviewed: Externally peer-reviewed. Financial Disclosure: The author declared that she did not receive any financial support during this article.

Supporting Institution

-

Project Number

-

Thanks

-

References

• 1. Baktir G. Wound Repair and Experimental Wound Models. Experimed. 2019;9:130-137.
• 2. Wang PH, Huang BS, Horng HC, et al. Wound healing. J Chin Med Assoc. 2018;81:94-101.
• 3. Wilkins RG, Unverdorben M. Wound cleaning and wound healing: a concise review. Adv Skin Wound Care. 2013;26:160-163.
• 4. Cano Sanchez M, Lancel S, Boulanger E, et al. Targeting Oxidative Stress and Mitochondrial Dysfunction in the Treatment of Impaired Wound Healing: A Systematic Review. Antioxidants (Basel). 2018;7:98.
• 5. Sung TJ, Wang YY, Liu KL, et al. Pholiota nameko Polysaccharides Promotes Cell Proliferation and Migration and Reduces ROS Content in H2O2-Induced L929 Cells. Antioxidants (Basel). 2020;9:65.
• 6. Gouin JP, Kiecolt-Glaser JK. The impact of psychological stress on wound healing: methods and mechanisms. Immunol Allergy Clin North Am. 2011;31:81-93.
• 7. Wang L, Feng T, Su Z, et al. Latest research progress on anticancer effect of baicalin and its aglycone baicalein. Arch Pharm Res. 2022;45:535-557.
• 8. Dinda B, Dinda S, DasSharma S, et al. Therapeutic potentials of baicalin and its aglycone, baicalein against inflammatory disorders. Eur J Med Chem. 2017;131:68-80.
• 9. Gasiorowski K, Lamer-Zarawska E, Leszek J, et al. Flavones from root of Scutellaria baicalensis Georgi: drugs of the future in neurodegeneration? CNS Neurol Disord Drug Targets. 2011;10:184-191. 10. Hu Z, Guan Y, Hu W, et al. An overview of pharmacological activities of baicalin and its aglycone baicalein: New insights into molecular mechanisms and signaling pathways. Iran J Basic Med Sci. 2022;25:14-26.
• 11. Bainbridge P. Wound healing and the role of fibroblasts. J Wound Care. 2013;22:407-408.
• 12. Yang JY, Li M, Zhang CL, et al. Pharmacological properties of baicalin on liver diseases: a narrative review. Pharmacol Rep. 2021;73:1230-1239.
• 13. Cui X, Sun X, Lu F, et al. Baicalein represses TGF-β1-induced fibroblast differentiation through the inhibition of miR-21. Toxicol Appl Pharmacol. 2018;358:35-42.
• 14. Cinar I. Evaluation of Protective Effects of Gossypin against Hydrogen Peroxide Damage in L929 Fibroblast Cells. Kafkas Journal of Medical Sciences. 2020;10:15-23.
• 15. Jussofie A, Kirsch M, de Groot H. Ca2+-dependent cytotoxicity of H2O2 in L929 cells: the role of H2O2-induced Na+-influx. Free Radic Biol Med. 1998;25:712-719.
• 16. Kang KA, Zhang R, Piao MJ, et al. Baicalein inhibits oxidative stress-induced cellular damage via antioxidant effects. Toxicol Ind Health. 2012;28:412- 421.
• 17. Kang KA, Piao MJ, Kim KC, et al. Fisetin attenuates hydrogen peroxideinduced cell damage by scavenging reactive oxygen species and activating protective functions of cellular glutathione system. In Vitro Cell Dev Biol Anim. 2014;50:66-74. 18. Mary VS, Theumer MG, Arias SL, et al. Reactive oxygen species sources and biomolecular oxidative damage induced by aflatoxin B1 and fumonisin B1 in rat spleen mononuclear cells. Toxicology. 2012;302:299-307.
• 19. Liu B, Jian Z, Li Q, et al. Baicalein protects human melanocytes from H₂O₂-induced apoptosis via inhibiting mitochondria-dependent caspase activation and the p38 MAPK pathway. Free Radic Biol Med. 2012;53:183-193.
• 20. Ransy C, Vaz C, Lombès A, et al. Use of H2O2 to Cause Oxidative Stress, the Catalase Issue. Int J Mol Sci. 2020;21:9149.
• 21. Karamese M, Erol HS, Albayrak M, et al. Anti-oxidant and anti-inflammatory effects of apigenin in a rat model of sepsis: an immunological, biochemical, and histopathological study. Immunopharmacol Immunotoxicol. 2016;38:228-237.
• 22. Younus H. Therapeutic potentials of superoxide dismutase. Int J Health Sci (Qassim). 2018;12:88-93. 23. Tan S, Zhou S, Luo Y. Baicalein pretreatment confers cardioprotection against acute myocardial infarction by activating the endothelial nitric oxide synthase signaling pathway and inhibiting oxidative stress. Mol Med Rep. 2014;9:2429-2434. 24. Ye F, Wang H, Zhang L, et alJ. Baicalein induces human osteosarcoma cell line MG-63 apoptosis via ROS-induced BNIP3 expression. Tumour Biol. 2015;36:4731-4740.
• 25. Lin L, Wu XD, Davey AK, et al. The anti-inflammatory effect of baicalin on hypoxia/reoxygenation and TNF-alpha induced injury in cultural rat cardiomyocytes. Phytother Res. 2010;24:429-437.
• 26. Cheng PY, Lee YM, Wu YS, et al. Protective effect of baicalein against endotoxic shock in rats in vivo and in vitro. Biochem Pharmacol. 2007;73:793-804.
• 27. Meng H, Fu G, Shen J, et al. Ameliorative Effect of Daidzein on Cisplatin- Induced Nephrotoxicity in Mice via Modulation of Inflammation, Oxidative Stress, and Cell Death. Oxid Med Cell Longev. 2017;2017:3140680.
• 28. Orzechowska B, Chaber R, Wiśniewska A, et al. Baicalin from the extract of Scutellaria baicalensis affects the innate immunity and apoptosis in leukocytes of children with acute lymphocytic leukemia. Int Immunopharmacol. 2014;23:558-567.