Ethanol Extract of Ganoderma lucidum Augments Cellular Anti-oxidant Defense through Activation of Nrf2/HO-1

Lee, Yoo-hwan,Kim, Jung-hee,Song, Choon-ho,Jang, Kyung-jeon,kim, Cheol-hong,Kang, Ji-Sook,Choi, Yung-hyun,Yoon, Hyun-Min KOREAN PHARMACOPUNCTURE INSTITUTE 2016 Journal of pharmacopuncture Vol.19 No.1

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

심실중격결손증-치험 1예-

조광현,우종수,황윤호,이양행,박철호,김종성,김철호,이경순,조영일,박태인 인제대학교 1985 仁濟醫學 Vol.6 No.4

A successful treatment of infants and children with congenital heart defects depends upon several factors: diagnosis must be accurate, the operative technique, including techniques of perfusion and myocardial preservation, must be suited to the needs of patients and intensive cares, including accurate anesthesia, must be applied through all stages of investigation and treatment, i.e. before, during and after the operation. Recently we performed a open heart surgery successfully for a six-years old girl who had suffered from frequent upper respiratory tract infection and mild to moderate exertional dyspnea with congenital heart defects (ventricular septal defect, Kirklin type II, with patent foramen ovale). The operation and postoperative course were very smooth. And now we report this with review of literatures.

Morin exerts cytoprotective effects against oxidative stress in C2C12 myoblasts via the upregulation of Nrf2-dependent HO-1 expression and the activation of the ERK pathway

Lee, Moon Hee,Han, Min Ho,Lee, Dae-Sung,Park, Cheol,Hong, Su-Hyun,Kim, Gi-Young,Hong, Sang Hoon,Song, Kyoung Seob,Choi, Il-Whan,Cha, Hee-Jae,Choi, Yung Hyun UNKNOWN 2017 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.39 No.2

<P>In the present study, we investigated the cytoprotective efficacy of morin, a natural flavonoid, against oxidative stress and elucidated the underlying mechanisms in C2C12 myoblasts. Our results indicated that morin treatment prior to hydrogen peroxide (H2O2) exposure significantly increased cell viability and prevented the generation of reactive oxygen species. H2O2-induced comet-like DNA formation and gamma H2AX phosphorylation were also markedly suppressed by morin with a parallel inhibition of apoptosis in C2C12 myoblasts, suggesting that morin prevented H2O2-induced cellular DNA damage. Furthermore, morin markedly enhanced the expression of heme oxygenase-1 (HO-1) associated with the induction and phosphorylation of nuclear factor-erythroid 2-related factor 2 (Nrf2) and the inhibition of Kelch-like ECH-associated protein 1 (Keapl) expression. Notably, these events were eliminated by transient transfection with Nrf2-specific small interfering RNA. Additional experiments demonstrated that the activation of the Nrf2/HO-1 pathway by morin was mediated by the extracellular signal-regulated kinase (ERK) signaling cascade. This phenomenon was confirmed with suppressed Nrf2 phosphorylation and consequently diminished HO-1 expression in cells treated with a pharmacological inhibitor of ERK. Collectively, these results demonstrated that morin augments the cellular antioxidant defense capacity through the activation of Nrf2/HO-1 signaling, which involves the activation of the ERK pathway, thereby protecting C2C12 myoblasts from H2O2,-induced oxidative cytotoxicity.</P>

Methanol extract of Myelophycus caespitosus ameliorates oxidative stress‑induced cytotoxicity in C2C12 murine myoblasts via activation of heme oxygenase-1

Cheol Park,Hyun Hwangbo,Han Min Ho,Jin-Woo Jeong,조승목,김기영,황혜진,Yung Hyun Choi 한국수산과학회 2023 Fisheries and Aquatic Sciences Vol.26 No.1

Myelophycus caespitosus, a brown alga belonging to genus Myelophycus, has been traditionally used as a food and medicinal resource in Northeastern Asia. However, few studies have been conducted on its pharmacological activity. In this study, we evaluated whether methanol extract of M. caespitosus (MEMC) could protect against oxidative damage caused by hydrogen peroxide (H2O2) in C2C12 murine myoblasts. Our results revealed that MEMC could suppress H2O2-induced growth inhibition and DNA damage while blocking the production of reactive oxygen species. In H2O2-treated cells, cell cycle progression was halted at the G2/M phase, accompanied by changes in expression of key cell cycle regulators. However, these effects were attenuated by MEMC. In addition, we found that MEMC protected cells from induction of apoptosis associated with mitochondrial impairment caused by H2O2 treatment. Furthermore, MEMC enhanced the phosphorylation of nuclear factor-erythroid-2 related factor 2 (Nrf2) and expression and activity of heme oxygenase-1 (HO-1) in H2O2-treaetd C2C12 myoblasts. However, such anti-apoptotic and cytoprotective effects of MEMC were greatly abolished by HO-1 inhibitor, suggesting that MEMC could increase Nrf2-mediated activity of HO-1 to protect C2C12 myoblasts from oxidative stress.

Antioxidant and cytoprotective effects of morin against hydrogen peroxide-induced oxidative stress are associated with the induction of Nrf-2-mediated HO-1 expression in V79-4 Chinese hamster lung fibroblasts

Lee, Moon Hee,Cha, Hee-Jae,Choi, Eun Ok,Han, Min Ho,Kim, Sung Ok,Kim, Gi-Young,Hong, Su Hyun,Park, Cheol,Moon, Sung-Kwon,Jeong, Soon-Jeong,Jeong, Moon-Jin,Kim, Wun-Jae,Choi, Yung Hyun Spandidos Publications 2017 International journal of molecular medicine Vol.39 No.3

<P>Natural phytochemicals of plant origin, including flavonoids, have been found to be potent antioxidants providing beneficial effects against oxidative stress-related diseases. The present study was carried out to investigate the antioxidant properties of morin, a flavonoid originally isolated from the flowering plants of the Moraceae family. Superoxide dismutase (SOD)-like activity and 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS(center dot+)) radical scavenging activity were determined. We also investigated the cytoprotective effects of morin against hydrogen peroxide (H2O2)-induced DNA damage and apoptosis in V79-4 Chinese hamster lung fibroblasts. Our results demonstrated that morin had strong scavenging effects against ABTS' radicals with enhanced SOD activity, which varied in a dose-dependent manner. Morin was found to reduce H2O2-induced intracellular reactive oxygen species generation and nuclear DNA damage, and it recovered cell viability damaged by H2O2 via inhibition of mitochondrial dysfunction-mediated apoptosis. Notably, the treatment of V79-4 cells with morin markedly enhanced the expression of heme oxygenase-1 (HO-1) but not quinone oxidoreductase-1, which was associated with the increased expression and phosphorylation of nuclear factor-erythroid 2-related factor 2 (Nrf2) and the downregulation of Kelch-like ECH-associated protein 1 expression. Based on our findings, we conclude that morin effectively ameliorated oxidative stress-induced DNA damage through intrinsic free radical scavenging activity and activation of the Nrf2/HO-1 pathway.</P>

진피 메탄올 추출물의 활성산소종 생성을 통한 인체 백혈병 세포의 apoptosis 유발

김가희(Ga Hee Kim),이문희(Moon Hee Lee),한민호(Min Ho Han),박철(Cheol Park),홍수현(Su Hyun Hong),최영현(Yung Hyun Choi) 한국생명과학회 2013 생명과학회지 Vol.23 No.8

진피(Citri Pericarpium)의 항암작용 기전 해석을 위하여 U937 백혈병 세포의 apoptosis 유발에 미치는 메탄올추출물(EMCP)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화, ROS의 생성 변화를 조사하였다. EMCP 처리에 의한 U937 세포의 증식 억제는 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 Flow cytometry 분석에 의한 Sub-G1기 세포 빈도의 증가로 확인하였다. EMCP 처리에 의한 apoptosis 유도에서 Bax 발현 증가, caspases의 활성 및 PARP의 단편화 등이 동반되었으며, ROS 생성의 증가와 연관성이 있었다. 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포 내 항산화 효소인 HO-1의 발현이 EMCP의 처리에 의해 증가되었으나 ROS 생성 억제제인 NAC의 전처리에 의해 감소된 HO-1의 발현은 전사인자인 Nrf2의 핵으로의 이동 억제와 관련되어 있었다. 이상의 결과에서 EMCP 처리에 의한 U937 세포의 apoptosis 유발에는 ROS 생성의 증가와 pNrf2에 의해 조절되는 HO-1의 발현 증가가 중요한 기전으로 작용한다는 것을 알 수 있었다. 이러한 자료는 진피의 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다. Citri Pericarpium is one of the most commonly used traditional herbal medicines in Korea, China, and Japan. Its extracts have many properties including the treatment of indigestion and inflammatory respiratory syndromes such as bronchitis and asthma. However, the underlying molecular mechanisms of anti-cancer activity and molecular targets are not fully understood. In this work, we investigated the anti-proliferative activity of Citri Pericapium (EMCP) methanol extract on reactive oxygen species (ROS) production and the association of these effects with apoptotic cell death using U937 human leukemia cells in vitro. EMCP treatment decreased cell proliferation in a dose-dependent manner following an increase of the sub-G1 phase, the down-regulation of Bax proteins, the activation of caspases, the degradation of poly (ADP-ribose) polymerase proteins (PARP), and the induction of ROS generation. However, the quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the EMCP-induced apoptosis effects. In addition, heme oxygenase-1 expression also recovered by inhibiting the nuclear translocation of phosphorylated NF-E2-related factor 2. Taken together, our data indicate that ROS are involved as key mediators in the early molecular events in the EMCP-induced apoptotic pathway.

케일 쥬스에 의한 AFLATOXIN B_1의 유전독성 억제 효과

최영현,박건영,이선미,유미애,이원호 부산대학교 유전공학연구소 1995 분자생물학 연구보 Vol.11 No.-

The inhibitory effect of the fresh juice of kale on the genotoxicity induced by aflatoxin B_1 (AFB_1) in Salmonella and Drosophila was investigated. The kale juice had strong inhibitory activity for His^- to His^+ reverse-mutations induced by AFB_1 acting on S. typhimurium TA100. A similar inhibitory effect was detected in somatic cell mutation assaying system of D. melanogaster. Using the wing hairs spot test, we found that the formation of mutant hairs in adult flies as a result of feeding with AFB_1 in their larval stages was efficiently inhibited by coadministration of the fresh juice of kale, which revealed that it can inhibit gene mutation, deletion and mitotic chromosomal recombination. These results seem to suggest that kale juice may exert its inhibitory effect to mutagenic and/or carcinogenic properties of DNA damaging agents.

결핵성 육아종에서 Thioredoxin peroxidase-2 의 발현

박근호,유형륜,정영진,윤기중,한원철,유대열,문형배 圓光大學校 醫科學硏究所 1999 圓光醫科學 Vol.15 No.2

Background: Thioredoxin peroxidase(TPX) is a kind of recently discovered antioxidant enzyme which react as rapid hydrogen ion donor for the removal of hydroperoxide. The action and distribution of the TPX was poorly understood in the human diseases. This experiments were designed for the study about the distribution of the TPX in the chronic granulomatous inflammation and about the correlation between the expression of TPX and the site of inflammation, histological activities of tuberculous inflammation or existence of mycobacterium in the inflammatory foci. Methods: The immunohistochemical stains were performed for the localization of the TPX-2 in the epithelioid cells, giant cells and lymphocytes in the chronic granulomatous inflammation. The tissue sections were obtained from the paraffin blocks of the 54 cases of tuberculosis (lung 21 cases, lymph node 12 cases, bone and soft tissue 12 cases, kidney 9 cases; active 33 cases, inactive 21 cases by the histologic classification; presence of mycobacterium 15 cases, no mycobacterium 39 cases by PCR reaction). Results: The expression of TPX-2 was 16.7% in the giant cells, 27.8% in the epithelioid cells and 100% in the lymphocytes of tuberculous inflammations. The expression of TPX-2 in the giant cells and epithelioid cells of the tuberculosis were 28.6% and 57.1% of the pulmonary tuberculosis; 33.3% in each cells of the renal tuberculosis; 0% in each cells of the lymph node or bone and soft tissue tuberculosis. The expression of TPX-2 in the giant cells and epithelioid cells were 9.1% in each cells of the active tuberculosis and were 28.6% and 57.1% in each cells of the inactive tuberculosis by histologic classification. The expression of TPX-2 in the giant cells and epithelioid cells was 40% in each cells of tuberculosis which mycobacteria were detected and the expression of TPX-2 was 7.7% and 23.1% in each cells which mycobacteria were not detected by PCR reaction in the paraffin embedded tissue. Conclusions: The above results were summarized that the TPX-2 in the giant cells and epithelioid cells were more frequently expressed in the inactive tuberculosis than in the active tuberculosis. These results suggest that the TPX-2 is a kind of regulating or suppressing factors in the activity of the tuberculosis.

再販賣價格維持制度 : 現行 法制의 問題點 및 改善策을 中心으로

金英鎬 동아대학교 법학연구소 1988 東亞法學 Vol.- No.7

술폰화 폴리스티렌-디비닐벤젠 혼성중합막을 이용한 유기산의 이동 특성에 관한 연구

조영일,한정우,선용호 연세대학교 대학원 1990 延世論叢 Vol.26 No.1

Abstract Initial fluxes of organic acids through a sulfonated polystyrene-divinylbenzene copolymer membrane (K-102, Asahi Kasei) were investigated by varying pH and initial concentration. In case of carboxylic acid such as formic acid and acetic acid, fluxes were higher when pH was lower than pKa and in case of amino acid such as L-alanine, flux was minimum when pH was isoelectic value. Initial fluxes have been found to vary in the range of 10-8-10-6mol/cm2 ㆍ s, depending on the kind of the acids, pH and initial concentrations. The relation between initial fluxes and initial concentrations has been expressed by Michaelis-Menten equation. For L-alanine when pH is 4.5, saturation constant, Km is 1.32 mol/L and maximum initial flux, Vmax is 2.31 mol/cm2 ㆍs