[논문]산화적 스트레스에 대한 석결명의 세포 보호 효과

김광연; 이승진; 지선영; 배수진; 송유림; 윤언정; 박선빈; 송종국; 손태진; 손재동; 김우현; 양주혜; 박선동; 김상찬; 김영우; 박광일

doi:10.14374/hfs.2021.29.2.71

산화적 스트레스에 대한 석결명의 세포 보호 효과
Cellular-protective effects of Nardotidis seu Sulculii Concha Extract against oxidative stress 원문보기

大韓韓醫學方劑學會誌 = Herbal formula science, v.29 no.2, 2021년, pp.71 - 80

김광연 (한국한의학연구원) , 이승진 (동국대학교 한의과대학 방제학교실) , 지선영 (대구한의대학교 한의학과) , 배수진 (동국대학교 한의과대학 방제학교실) , 송유림 (동국대학교 한의과대학 방제학교실) , 윤언정 (동국대학교 한의과대학 방제학교실) , 박선빈 (동국대학교 한의과대학 방제학교실) , 송종국 (용인대학교) , 손태진 (경일대학교) , 손재동 (경상대학교 수의과 대학) , 김우현 (경상대학교 수의과 대학) , 양주혜 (한국한의학연구원) , 박선동 (동국대학교 한의과대학 방제학교실) , 김상찬 (대구한의대학교 한의학과) , 김영우 (동국대학교 한의과대학 방제학교실) , 박광일 (경상대학교 수의과 대학)

Abstract ▼ AI-Helper

Objectives : This study investigated cellular-protective effects of Nardotidis seu Sulculii Concha water extract (NSCE) against oxidative stress induced by arachidonic acid (AA)+iron or tert-butylhydroperoxide (tBHP). Methods : In vitro, MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of AMP-activated kinase (AMPK) signaling pathway and autophagy related proteins. In vivo, mice were orally administrated with the aqueous extract of NSCE of 500 mg/kg for 3 days, and then injected with CCl4 0.5 mg/kg body weight to induce acute damage. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) analysis. Results : Treatment with NSCE inhibited cell death induced by AA+iron and tBHP. NSCE induced the phosphorylation of AMPK, and this compound also induced the phosphorylation of LKB1, an upstream kinase of AMPK, and Acetyl-CoA carboxylase (ACC), a primary downstream target of AMPK. NSCE increased the protein levels of autophagic markers (LC3II and beclin-1) and decreased the phosphorylation of mammalian target of rapamycin (mTOR) and simultaneously increased the phosphorylation of unc-51-like kinase-1 (ULK-1) in time-dependent manner. Conclusions : NSCE has the ability 1) to protect cells against oxidative stress induced by AA+iron or tBHP. NSCE 2) to activate AMP-activated protein kinase (AMPK), and 3) to regulate autophagy, an important regulator in cell survival.

주제어

표/그림 (4)

그림 Figure 1. Effects of Nardotidis seu Sulculii Concha water extract (NSCE) on cell viability of HepG2 cells. (A) Cell viability of NSCE in AA+Fe-treated HepG2 cells. Cells were pretreated with various concentrations (30–1000 μg/ml) of NSCE for 1 h and then treated with 30 μM arachidonic acid (AA) for 12 h, followed by treatment of 5 μM iron for 2 h. (B) Cell viability of NSCE in tBHP-treated HepG2 cells. Cells were pretreated with various concentrations (30–1000 μg/ml) of NSCE for 1 h and then followed by treated with 200 μM tBHP for 24 h. Cell viability was determined using a Cell Counting Kit-8. Cell viability is represented as the percentage relative absorbance compared with the controls. The results represent the mean ± standard error of the mean values of each cell in the same group. ###p < 0.001 versus the control group, *p < 0.05, **p < 0.01, ***p < 0.001 versus the AA+iron or tBHP-treated group.
그림 Figure 2. Effects of Nardotidis seu Sulculii Concha water extract (NSCE) on AMPK signaling pathway. (A) Effects of NSCE on AMPK, ACC and LKB1 expression in HepG2 cells. Phosphorylated form of AMPK, ACC and LKB1 were immunoblotted in the whole cell lysates treated with 300 μg/ml of NSCE for the indicated time. (B) Fold change of p-AMPKα protein expression was analysed by β-actin loading control. The results represent the mean ± standard error of the mean values of each mouse in the same group. * p < 0.05 versus the control group.
그림 Figure 3. Effects of Nardotidis seu Sulculii Concha water extract (NSCE) on autophagy-related protein expression. Effects of NSCE on LC3B, Beclin-1, p-mTOR and p-ULK1 expression in HepG2 cells. Protein expression of LC3B, Beclin-1, p-mTOR and p-ULK1 were immunoblotted in the whole cell lysates treated with 300 μg/ml of NSCE for the indicated time.
그림 Figure 4. Effects of Nardotidis seu Sulculii Concha water extract (NSCE) on CCl₄-induced hepatic damage in mice. (A) Effects of NSCE on serum AST level. (B) Effects of NSCE on serum ALT level. (C) Effects of NSCE on serum LDH level. The results represent the mean ± standard error of the mean values of each mouse in the same group. ###p < 0.001 versus the control group, ***p < 0.001 versus the CCl₄ alone.

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