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파라페닐렌디아민으로 세포 독성 및 염증을 유도한 마우스 대식세포에서 Derma GenieTM-H002의 세포 보호 및 항염증 효과

Cytoprotective and Anti-inflammatory Effects of Derma Genie™-H002 against Paraphenylenediamine-induced Cytotoxicity and Inflammation in RAW 264.7 Cells

김지선|조수현|김희수|안성관|배승희|신희재|정석윤|백세열|박석묵|임예진
한국피부과학연구원 (Korea Institute of Dermatological Sciences)|2024.06|아시안뷰티화장품학술지 (Asian Journal of Beauty and Cosmetology)|제22권 제2호|pp. 211-224

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참고문헌

키워드

세포독성항염증알레르기성 접촉 피부염염모제파라페닐렌디아민过敏性接触性皮炎，抗炎，细胞毒性，染发剂，对苯二胺
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초록

목적: 본 연구는 마우스 대식 세포 내에서 파라페닐렌디아민에 의한 세포 독성 및 염증 모델을 수립하고, 해당 in vitro 모델 내에서 5종의 생약 복합 추출물로 구성된 Derma GenieTM-H002의 세포 보호 및 항염 효능을 확인하고자 하였다. 방법: PPD에 의한 세포 독성 및 염증 모델에서 Derma GenieTM-H002의 효능을 평가하기 위해 수용성 테트라졸리움 염색법, 크리스탈 바이올렛 염색법, 그리고 젖산 탈수소효소 분석 시험을 포함한 다양한 생화학 분석법이 활용되었다. 또한 세포 사멸 연관 유전자 및 염증 연관 유전자들의 분석을 위해서 역전사 중합효소 연쇄 반응과 웨스턴 블롯이 활용되었다. 결과: Derma GenieTM-H002는 마우스 대식세포에서 유의한 세포 독성을 나타내지 않았다. 반면 PPD는 저농도(25 μM) 이상에서 유의한 세포 독성이 관찰되었다. PPD와 Derma GenieTM-H002의 병용 처리는 PPD에 의해 감소된 세포 생존율을 개선하였고, PPD에 의해 증가된 세포 독성을 완화하였으며, 세포 사멸을 촉진하는 단백질인 Bax의 발현량은 감소시키고, 세포 사멸을 억제하는 단백질인 Bcl-2의 발현량은 증가시킴으로써 PPD에 의한 독성 모델에서의 세포 보호 효능을 입증하였다. 또한 PPD와 Derma GenieTM-H002의 병용 처리는 PPD에 의해 증가된 염증 복합체 연관 유전자, 염증성 사이토카인 및 알레르기성 접촉 피부염 연관 유전자의 발현을 모두 감소시켰으며, PPD에 의해 증가된 NF-κB 단백질 발현량을 감소시켰으므로 PPD에 의한 염증 모델에서의 항염 효능을 입증하였다. 결론: 본 연구는 Derma GenieTM-H002가 PPD에 의해 유발된 마우스 대식 세포 내 독성 및 염증을 효과적으로 완화시킨다는 것을 입증하였다. 따라서 천연물 유래 원료인 Derma GenieTM-H002의 기능성 항염 소재로의 적용 가능성을 확인하였다.

Abstract

Purpose: This study aimed to establish an in vitro model to examine paraphenylenediamine (PPD)-induced cytotoxicity and inflammation within RAW 264.7 cells. Additionally, we investigated the cytoprotective and anti-inflammatory effects of Derma Genie™-H002 derived from extracts of five medicinal herbs. Methods: Various biochemical analyses, including water-soluble tetrazolium salt, crystal violet staining, and lactate dehydrogenase leakage assays, were employed to evaluate the impact of Derma Genie™-H002 on PPD-induced cytotoxicity and inflammation. Furthermore, apoptosis- and inflammation-related gene expression was analyzed using reverse transcription-polymerase chain reaction and Western blotting. Results: Derma Genie™-H002 exhibited no significant cytotoxicity in RAW 264.7 cells, whereas PPD induced notable cytotoxicity at concentrations >25 μM. Cotreatment with PPD and Derma Genie™-H002 not only improved cell proliferation rates but also alleviated cell viability in PPD-induced RAW 264.7 cells. Furthermore, findings demonstrated that Derma Genie™-H002 mitigates the cytotoxicity of PPD-induced RAW 264.7 cells by reducing the expression of the proapoptotic protein Bax and increasing the expression of the antiapoptotic protein Bcl-2. Moreover, cotreatment with PPD and Derma Genie™-H002 downregulated the mRNA expression of inflammasome-related genes, inflammatory cytokines, and allergic contact dermatitis-related genes, all elevated by PPD. The increased expression of nuclear factor-κB protein induced by PPD was also reduced upon cotreatment with Derma Genie™-H002, providing compelling evidence of its anti-inflammatory efficacy in the PPD-induced inflammation model. Conclusion: This study demonstrated that Derma Genie™-H002 effectively alleviates PPD-induced cytotoxicity and inflammation in RAW 264.7 cells. Therefore, the potential application of Derma Genie™-H002 as a naturally derived functional anti-inflammatory material has been validated.

참고문헌 (44)

  1. Adair K, "Drug hapten-specific T-cell activation : current status and unanswered questions", Proteomics, 21, pp.2000267~ (2021)
  2. Antonsson B, "The Bcl-2 protein family", Experimental Cell Research, 256, pp.50~57 (2000)
  3. Atri C, "Role of human macrophage polarization in inflammation during infectious diseases", International Journal of Molecular Science, 19, pp.1801~ (2018)
  4. Belsito DV, "The diagnostic evaluation, treatment, and prevention of allergic contact dermatitis in the new millennium", The Journal of Allergy and Clinical Immunology, 105, pp.409~420 (2000)
  5. Beltrani VS, "Contact dermatitis", Annals of Allergy, Asthma & Immunology, 78, pp.160~173 (1997)
  6. Berridge MV, "Tetrazolium dyes as tools in cell biology : new insights into their cellular reduction", Biotechnology Annual Review, 11, pp.127~152 (2005)
  7. Brites GS, "Allergic contact dermatitis : from pathophysiology to development of new preventive strategies", Pharmacological Research, 162, pp.105282~ (2020)
  8. Chaudhary SC, "Paraphenylenediamine poisoning", Nigerian Journal of Clinical Practice, 16, pp.258~259 (2013)
  9. Cohen DE., "Contact dermatitis: a quarter century perspective", Journal of the American Academy of Dermatology, 51, pp.S60~S63 (2004)
  10. Darwish RS, "Chemical profiling and identification of anti-inflammatory biomarkers of oriental Thuja(Platycladus orientalis)using UPLC", MS, Natural Product Research, pp.2021, 36[DOI]
  11. Erkes DA, "Hapten-induced contact hypersensitivity, autoimmune reactions, and tumor regression : plausibility of mediating antitumor immunity", Journal of Immunology Research, 2014, pp.175265~ (2014)
  12. Feoktistova M, "Crystal violet assay for determining viability of cultured cells", Cold Spring Harbor Protocol, pp.pdb.prot087379~ (2016)
  13. Fotakis G, "In vitro cytotoxicity assays : comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride", Toxicology Letters, 160, pp.171~177 (2006)
  14. Gan DL, "Volatile oil of Platycladus orientalis (L.) franco leaves exerts strong anti-inflammatory effects via inhibiting the IκB", NF-κB pathway, 2021, pp.41, 180~186
  15. Hamann D, "p-Phenylenediamine and other allergens in hair dye products in the United States : a consumer exposure study", Contact Dermatitis, 70, pp.213~218 (2014)
  16. Hamdouk M, "The association between prolonged occupational exposure to paraphenylenediamine(hairdye)and renal impairment", Arab Journal of Nephrology Transplantation, 4, pp.21~25 (2011)
  17. Handa S, "Contact dermatitis to hair dye : an update", Indian Journal of Dermatology, Venereology and Leprology, 78, pp.583~590 (2012)
  18. He Z, "A review of pharmacokinetic and pharmacological properties of asiaticoside, a major active constituent of Centella asiatica (L.) Urb", Journal of Ethnopharmacology, 302, pp.115865~ (2023)
  19. Kaplan DH, "Early immune events in the induction of allergic contact dermatitis", Immunology, 12, pp.114~124 (2012)
  20. Kim Y, "4, 5-Di-O-caffeoylquinic acid from Ligularia fischeri suppresses inflammatory responses through TRPV1 activation", Phytotherapy Research, 31, pp.1564~1570 (2017)
  21. Kostner L, "Allergic contact dermatitis", Immunology and Allergy Clinical North America, 37, pp.141~152 (2017)
  22. Kupcsik L., "Mammalian cell viability: methods and protocols", pp.13~19 (2011), Humana Press
  23. Lawrence T, "The nuclear factor NF-κB pathway in inflammation", Cold Spring Harbor Perspectives in Biology, 1, pp.a001651~ (2009)
  24. Lee KH, "Analgesic and anti-inflammatory effects of Ligularia fischeri leaves in experimental animals", Journal of Ethnopharmacology, 120, pp.103~107 (2008)
  25. Leonard A, "The unique molecular signatures of contact dermatitis and implications for treatment", Clinical Reviews in Allergy & Immunology, 56, pp.1~8 (2019)
  26. Li Y, "Contact dermatitis : classifications and management", Clinical Reviews in Allergy & Immunology, 61, pp.245~281 (2021)
  27. Lin P, "Centella asiatica alleviates psoriasis through JAK", STAT3-mediated inflammation : an in vitro and in vivo study, 2023, pp.317, 116746~
  28. Mark BJ, "Allergic contact dermatitis", The Medical Clinical of North America, 90, pp.169~185 (2006)
  29. McFadden JP, "Clinical and experimental aspects of allergic contact dermatitis to paraphenylenediamine", Clinics in Dermatology, 29, pp.316~324 (2011)
  30. Mills C, "M1 and M2 macrophages : oracles of health and disease", Critical Reviews in Immunology, 32, pp.463~488 (2012)
  31. Morel LJF, "A methanolic extract of Eclipta prostrata (L.) L. decreases inflammation in a murine model of chronic allergic asthma via inhibition of the NF-kappa-B pathway", Journal of Ethnopharmacology, 318, pp.116930~ (2024)
  32. Mukkanna KS, "Para-phenylenediamine allergy : current perspectives on diagnosis and management", Journal of Asthma and Allergy, 10, pp.9~15 (2017)
  33. Nograles KE, "IL-22–producing"T22"T cells account for upregulated IL-22 in atopic dermatitis despite reduced IL-17–producing TH17 T cells", The Journal of Allergy and Clinical Immunology, 123, pp.1244~1252 (2009)
  34. Novak-Bilić G, "Irritant and allergic contact dermatitis–skin lesion characteristics", Acta Clinica Croatica, 57, pp.713~719 (2018)
  35. Oakes T, "The T cell response to the contact sensitizer paraphenylenediamine is characterized by a polyclonal diverse repertoire of antigen-specific receptors", Frontiers in Immunology, 8, pp.162~ (2017)
  36. Reed JC, "Bcl-2 family proteins", Oncogene, 17, pp.3225~3236 (1998)
  37. Scarcello E, "Mind your assays: misleading cytotoxicity with the WST- 1 assay in the presence of manganese", PLoS One, 15, pp.e0231634~ (2020)
  38. Tasneem S, "Molecular pharmacology of inflammation : Medicinal plants as antiinflammatory agents", Pharmacological Research, 139, pp.126~140 (2019)
  39. Thangaraj P., "Pharmacological assays of plant-based natural products", pp.159~161 (2016), Springer International Publishing
  40. Thyssen JP, "p-Phenylenediamine sensitization is more prevalent in central and southern European patch test centres than in Scandinavian : results from a multicentre study", Contact Dermatitis, 60, pp.314~319 (2009)
  41. Van Belle AB, "IL-24 contributes to skin inflammation in Para-Phenylenediamine-induced contact hypersensitivity", Scientic Reports, 9, pp.1852~ (2019)
  42. Woo H, "Rhus semialata M. extract ameliorate para-phenylenediamineinduced toxicity in keratinocytes", Toxicology Reports, 8, pp.96~105 (2021)
  43. Yang Y, "Recent advances in the mechanisms of NLRP3 inflammasome activation and its inhibitors", Cell Death & Disease, 10, pp.128~ (2019)
  44. Zug KA, "Patch-test results of the North American contact dermatitis group 2005-2006", Dermatitis, 20, pp.149~160 (2009)

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