[논문]산화스트레스를 유발한 위 상피세포에서 수치 치자의 세포 보호 효과

이종록; 김상찬; 박숙자

doi:10.6116/kjh.2023.38.1.21.

산화스트레스를 유발한 위 상피세포에서 수치 치자의 세포 보호 효과
Cytoprotective effect of the processed Gardeniae Fructus on oxidative stress-induced gastric epithelial cells 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.38 no.1, 2023년, pp.21 - 30

이종록 (대구한의대학교 제약공학과) , 김상찬 (대구한의대학교 한의과대학) , 박숙자 (대구한의대학교 제약공학과)

Abstract ▼ AI-Helper

Objective : Gardeniae Fructus (GF) has bitter and cold nature. Thus, it has been traditionally prescribed in processed form roasted with ginger juice for patients with a weak stomach. This study investigated the effects of processed GF in tert-butyl hydroperoxide (tBHP)-treated gastric epithelial cells. Methods : Processed GF was made by applying 40% ginger juice or 10% ethanol for 24 h and then roasting at 150℃ for 5 minutes. Apoptosis was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Mitochondrial membrane potential (MMP) was monitored by flow cytometry using the membrane permeable fluorescent dye Rh123. Protein expression was measured by Western blot analysis. Results : Cell viability was reduced by tBHP and restored by ethanol extract of GF (GFE). In the TUNEL assay, it was found that cell death by tBHP was due to apoptosis, and GFE had an anti-apoptotic effect. Processed GF roasted with ginger juice showed the best anti-apoptotic effect. Processed GF also inhibited MMP loss and restored tBHP-induced changes in expression levels of apoptosis-related proteins. Increased ROS production and GSH depletion after tBHP treatment were significantly reduced by processed GF. In addition, tBHP-induced activation of mitogen-activated protein kinase (MAPK) was inhibited by processed GF. Conclusion : These results demonstrate that the processed GF is able to protect gastric epithelial cells from oxidative stress-induced cell death with antiapoptotic and antioxidant activity. In addition, it shows that the processing of GF, which have been traditionally used for gastrointestinal protection, partially have scientific validity.

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표/그림 (6)

그림 Fig. 1. Effect of GFE on tBHP-induced cytotoxicity in AGS cells Cells were treated with GFE (50 ㎍/㎖) for 24 h, and then cultured with or without tBHP (500 μM) for 3 h. Cell viability was determined by MTT assay. Values are expressed as the mean ± SD. The different letters (a-d) indicate significant differences (p < 0.05).
그림 Fig. 2. Effect of GFE on tBHP-induced apoptosis in AGS cells (A) Apoptotic cells were confirmed by TUNEL staining, and arrows indicate TUNEL-positive cells. (B) The number of TUNEL-positive cells were counted and expressed as a fold value of control. Values are expressed as the mean ± SD. The different letters (a-d) indicate significant differences (p < 0.05).
그림 Fig. 3. Effect of GFE on the expression of apoptosis-associated proteins Cells were treated with GFE (50 ㎍/㎖) for 24 h, and then cultured with or without tBHP (500 μM) for 3 h. The level of expression of the apoptosis marker protein was monitored by Western blot analysis. Actin was used as a loading control. Bar graphs show the band density of Western blot. Data represent the mean ± SD of three separate experiments. The different letters (a-c) indicate significant differences (p < 0.05).
$Fig. 5. Effects of GFE on mitochondria membrane potential (MMP) (A) MMP was measured by flow cytometry using Rh123. (B) M1 fraction with low Rh123 fluorescence intensity was counted as an indication of MMP destruction. Values are expressed as the mean ± SD. The different letters (a-c) indicate significant differences (p < 0.05).$ 그림 Fig. 5. Effects of GFE on mitochondria membrane potential (MMP) (A) MMP was measured by flow cytometry using Rh123. (B) M1 fraction with low Rh123 fluorescence intensity was counted as an indication of MMP destruction. Values are expressed as the mean ± SD. The different letters (a-c) indicate significant differences (p < 0.05).
그림 Fig. 4. Effect of GFE on the levels of ROS and GSH (A) ROS level was measured after DCF-DA staining for 15 min and the fluorescence intensity was expressed as percentage of control. (B) GSH contents were measured in cell homogenates as described in material and method section. Values are expressed as the mean ± SD. The different letters (a-d) indicate significant differences (p < 0.05).
그림 Fig. 6. Effect of GFE on the expression of MAPK protein Expression of phosphorylated MAPK protein was monitored by Western blot analysis after 24 h incubation of GFE and 30 min exposure of tBHP. Actin was measured as an internal control. Bar graphs show the band density of Western blot. Data represent the mean ± SD of three separate experiments. The different letters (a-c) indicate significant differences (p < 0.05).

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저자의 다른 논문 :

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동의어: Packet Collision Rate

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