[논문]HaCaT 세포의 산화 스트레스로 인한 세포자멸사에서 정향의 보호효과

박숙자

doi:10.6116/kjh.2021.36.5.93.

HaCaT 세포의 산화 스트레스로 인한 세포자멸사에서 정향의 보호효과
Protective effect of Caryophylli Flos on apoptosis caused by oxidative stress in HaCaT cells 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.36 no.5, 2021년, pp.93 - 99

Abstract ▼ AI-Helper

Objective : Caryophylli Flos has been used in Korean medicine to relieve vomiting and pains caused by chills that make fluid circulation difficult. This study was designed to investigate the protective effect of ethanol extract of Caryophylli Flos (CF) in hydrogen peroxide (H2O2)-induced apoptotic cell death in human keratinocyte HaCaT cells. Methods : CF was prepared by extracting 200 g of Caryophylli Flos in 2 L of ethanol for 48 h. Cell viability was measured by MTT assay, and the protein expression was monitored by Western blot analysis. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Reactive oxygen species (ROS) was measured using fluorescent dye, and reduced glutathione (GSH) was determined with a colorimetric commercial kit. Results : CF protected HaCaT cells from cell death caused by oxidative stress after H2O2 treatment. H2O2 amplified generation of ROS and induced depletion of GSH, whereas these changes in ROS and GSH were inhibited by GF treatment. In addition, H2O2 resulted in apoptosis as assessed by TUNEL assay and the expression of apoptosis regulator proteins. However, cells treated with CF showed a decrease in TUNEL-positive cells and restored the reduced expression of procaspase-9, -3 and PARP. Conclusion : This study showed cytoprotective effects of CF by anti-apoptotic activity while exerting antioxidative activity in H2O2-treated HaCaT cells. These results suggest that CF could be beneficial in skin damage caused by oxidative stress.

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표/그림 (5)

그림 Fig. 1. Effect of CF on H₂O₂-induced cytotoxicity in HaCaT cells (A) Cells were treated with various concentrations (10, 30, 100 ㎍/㎖) of CF for 24 h to determine the optimal experimental concentration. (B) HaCaT cells were pre-treated with CF for 1 h and then exposed with 500 μM of H₂O₂ for an additional 18 h. C, control; ns, not significant; ##p < 0.01, significant as compared to control; **p ＜ 0.01, *p ＜ 0.05, significant as compared to H₂O₂ alone.
그림 Fig. 2. Effect of CF on H₂O₂-induced ROS production HaCaT cells were pre-treated with CF for 1 h and then exposed with H₂O₂ for an additional 18 h. ROS level was measured after DCF-DA staining for 15 min and the fluorescence intensity was expressed as percentage of control. C, control; ##p < 0.01, significant as compared to control; **p ＜ 0.01, significant as compared to H₂O₂ alone.
그림 Fig. 3. Effect of CF on GSH levels altered by H₂O₂ treatment HaCaT cells were pre-treated with CF for 1 h and then exposed with H₂O₂ for an additional 18 h. GSH contents were measured in cell homogenates as described in material and method section. Data were expressed as percentage of control. C, control; ##p < 0.01, significant as compared to control; **p ＜ 0.01, significant as compared to H₂O₂ alone.
그림 Fig. 4. Effect of CF on H₂O₂-induced apoptosis in HaCaT cells (A) Apoptotic cells were detected by TUNEL assay. Arrows indicate TUNEL-positive cells with dark brown color. (B) TUNEL-positive cells were counted, and the results were expressed as the fold value of control. C, control; ##p < 0.01, significant as compared to control; **p ＜ 0.01, significant as compared to H₂O₂ alone.
그림 Fig. 5. Effect of CF on the expression of apoptosis-associated proteins (A) HaCaT cells were pre-treated with CF for 1 h and then exposed with H₂O₂ for an additional 18 h. The level of expression of the apoptosis marker protein was monitored by Western blot analysis. β-actin was used as a loading control. (B) The relative intensity of protein band was measured by scanning densitometry and quantified as a fold value of control. C, control; ##p < 0.01, significant as compared to control; **p ＜ 0.01, *p ＜ 0.05, significant as compared to H₂O₂ alone.

참고문헌 (29)

Oriental Pharmacology Committee. Oriental Pharmacology. 2nd ed. Seoul : Shinil. 2006:608-10.
Huh J. Donguibogam. 1st ed. Bubin Publishers. 2005:3588.
Chaieb K, Zmantar T, Ksouri R, Hajlaoui H, Mahdouani K, Abdelly C, Bakhrouf A. Antioxidant properties of the essential oil of Eugenia caryophyllata and its antifungal activity against a large number of clinical Candida species. Mycoses. 2007;50(5):403-6.

상세보기
Son KH. Constituents from Syzygium aromaticum Merr. et. Perry. Nat Prod Sci. 1998;4(4):263-7.
Youn SN, Kim YJ, Kim MR, Yoo WK. Anti-oxidant effect of ethanol extract from mixture including Caryophylli Flos, Aucklandiae Radix and Angelicae Dahuricae Radix. Kor J Herbol. 2018;33(6):1-8.

원문보기 상세보기
Lee OH, Jung SH, Song JY. Antimicrobial activity of clove extract by extraction solvents. J Korean Soc Food Sci Nutr. 2004;33(3):494-9.

원문보기 상세보기
Gowda NKS, Malathi V, Suganthi RU. Effect of some chemical and herbal compounds on growth of Aspergillus parasiticus and aflatoxin production. Anim Feed Sci Technol. 2004;116:281-91.

상세보기
Lee KY, Kang DH, Kim HT. The Effects of Syzygium aromaticum extract Spread on the Allergic Contact Dermatitis induced by DNCB. J Korean Med Ophthalmol Otolaryngol Dermatol. 2013;26(4):1-14

원문보기 상세보기
Prasad RC, Herzog B, Boone B, Sims L, WaltnerLaw M. An extract of Syzygium aromaticum represses genes encoding hepatic gluconeogenic enzymes. J Ethnopharmacol. 2005;96:295-301.

상세보기
Yang YY, Lee MJ, Lee HS, Park WH. Screening of antioxidative, anti-platelet aggregation and antithrombotic effects of clove extracts. J Physiol & Pathol Korean Med. 2011;25(3):471-81.
Slominski A, Tobin DJ, Shibahara S, Wortsman J. Melanin pigmentation in mammalian skin and its hormonal regulation. Physiol Rev. 2004;84(4):1155-228.

상세보기
Glady A, Tanaka M, Moniaga CS, Yasui M, HaraChikuma M. Involvement of NADPH oxidase 1 in UVB-induced cell signaling and cytotoxicity in human keratinocytes. Biochem Biophys Rep. 2018;14:7-15.
Mizutani T, Sumida H, Sagawa Y, Okano Y, Masaki H. Carbonylated proteins exposed to UVA and to blue light generate reactive oxygen species through a type I photosensitizing reaction. J Dermatol Sci. 2016;84(3):314-21.

상세보기
Andersson H, Piras E, Demma J, Hellman B, Brittebo E. Low levels of the air pollutant 1-nitropyrene induce DNA damage, increased levels of reactive oxygen species and endoplasmic reticulum stress in human endothelial cells. Toxicology. 2009;262(1):57-64.

상세보기
McDaniel D, Farris P, Valacchi G. Atmospheric skin aging-Contributors and inhibitors. J Cosmet Dermatol. 2018;17(2):124-37.

상세보기
Kim JK, Park SM, Jegal KH, Kim YW, Byun SH, Kim SC, Cho IJ. Gleditsia Spina extract protects hepatocytes from oxidative stress through Nrf2 activation. Kor J Herbol. 2015;30(4):57-64.

원문보기 상세보기
Briganti S, and Picardo M. Antioxidant activity, lipid peroxidation and skin diseases. What's new. J Eur Acad Dermatol Venereol. 2003;17:663-9.

상세보기
Pourzand C, Tyrrell RM. Apoptosis, the role of oxidative stress and the example of solar UV radiation. Photochem Photobiol. 1999;70:380-90.

상세보기
Los M, Mozoluk M, Ferrari D, Stepczynska A, Stroh C, Renz A. Herceg Z, Wang ZQ, Schulze-Osthoff K. Activation and caspase-mediated inhibition of PARP: a molecular switch between fibroblast necrosis and apoptosis in death receptor signaling. Mol Biol Cell. 2002;13(3):978-88.

상세보기
Pelle E, Mammone T, Maes D, Frenkel K. Keratinocytes act as a source of reactive oxygen species by transferring hydrogen peroxide to melanocytes. J Investig Dermatol. 2005;124(4): 793-7.

상세보기
Seo SH, Choi MO. protective effects of Lonicerae Japonicae Flos against hydrogen peroxidase-induced oxidative stress on human keratinocyte, HaCaT cells. Kor J Herbol. 2013;28(4):57-62.

원문보기 상세보기
Zuliani T, Denis V, Noblesse E, Schnebert S, Andre P, Dumas M, Ratinaud MH. Hydrogen peroxide-induced cell death in normal human keratinocytes is differentiation dependent. Free Radic Biol Med. 2005;38(3):307-16.

상세보기
Mohania D, Chandel S, Kumar P, Verma V, Digvijay K, Tripathi D, Choudhury K, Mitten SK, Shah D. Ultraviolet radiations: Skin defense-damage mechanism. Adv Exp Med Biol. 2017;996:71-87.
Bito T, Nishigori C. Impact of reactive oxygen species on keratinocyte signaling pathways. J Dermatol Sci. 2012;68(1):3-8.

상세보기
Masaki H, Okano Y, Ochiai Y, Obayashi K, Akamatsu H, Sakurai H. Alpha-tocopherol increases the intracellular glutathione level in HaCaT keratinocytes. Free Radic Res. 2002;36(6):705-9.

상세보기
Kyrylkova K, Kyryachenko S, Leid M, Kioussi C. Detection of apoptosis by TUNEL assay. Methods Mol Biol. 2012;887:41-7.

상세보기
Noh GP, Byun SH, Lee JR, Park SJ, Kim SC. Protective effect of Citrus unshiu peel on the cadmium-induced apoptosis in HepG2 cells. Kor J Herbol. 2021;36(1):41-9.
Li JL, Liu N, Chen XH, Sun M, Wang CB. Inhibition of UVA-induced apoptotic signaling pathway by polypeptide from Chlamys farreri in human HaCaT keratinocytes. Radiat Environ Biophys. 2007;46(3):263-8.

상세보기
Lyakhovich A, Surralles J. Constitutive activation of caspase-3 and Poly ADP ribose polymerase cleavage in fanconi anemia cells. Mol Cancer Res. 2010;8(1):46-56.

상세보기

저자의 다른 논문 :

표제어: PCR

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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