Menadione (Vitamin K3) decreases melanin synthesis through ERK activation in Mel-Ab cells

Kim, E.H.,Kim, M.K.,Yun, H.Y.,Baek, K.J.,Kwon, N.S.,Park, K.C.,Kim, D.S. North-Holland ; Elsevier Science Ltd 2013 european journal of pharmacology Vol.718 No.1

Menadione is a synthetic vitamin K3 derivative. Here, we examined the effects of menadione on melanogenesis and its related signaling pathways. Our results showed that melanin content was significantly reduced after menadione treatment in a dose-dependent manner. However, menadione treatment did not reduce tyrosinase activity directly. Wnt signaling is known to play a major role in the control of melanin synthesis. Thus, we tested the effects of menadione treatment on GSK3β and β-catenin signaling, but found that menadione did not influence either of these signaling pathways. We also investigated changes in the phosphorylation of ERK, which is related to melanin regulation. These results indicated that menadione treatment led to the phosphorylation of ERK. Additionally, menadione treatment reduced both MITF and tyrosinase protein levels. Treatment with PD98059, a specific ERK pathway inhibitor, restored menadione-induced melanin reduction and also prevented MITF and tyrosinase downregulation by menadione. These results suggest that the hypopigmentary action of menadione is due to MITF and tyrosinase downregulation by ERK activation.

Menadione의 대사체인 Menadione-Glutathione Conjugate(MEN-SG)가 흰쥐 혈소판에 미치는 세포독성의 평가 및 MEN-SG의 안정성에 관한 연구

서동철,정선화,이주영,김미정,정진호,Seo, Dong-Chul,Chung, Sun-Hwa,Lee, Joo-Young,Kim, Mee-Jeong,Chung, Jin-Ho 한국독성학회 1995 Toxicological Research Vol.11 No.2

Menadione-ghitathione conjugate (MEN-SG), a metabolite of menadione, is known to be a redoxcycler in rat hepatocyte subcellular fraction. Therefore, it was assumed that MEN-SG could exert cytotoxlclty to ral platelets, another target tissue of menadione. We first synthesized MEN-SG, the identity of which was verified by mass, $^1{H}$-NMR and UV-visible spectra. In addition, the stability of MEN-SG was investigated in biological assay system. MEN-SG was degraded in a time-dependent manner in DMSO which had been used as a vehicle and thus, tris-HCl buffer was used as a vehicle of MEN-SG despite the low solubility in it. Perchloric acid as well as platelets itself did not affect the stability of MEN-SG. Our next attempt was the evaluation of cytotoxicity of MEN-SG in rat platelets. MEN-SG did not induce cytotoxicity to rat platelets measured by two different methods, LDH release and turbidity changes. The extents of oxygen consumption by MEN-SG in intact platelets were significantly lower than those by menadione, though it had been observed that oxygen consumptions by menadione and MENSG were similar in subcellular fractioas of platelets. These results suggest that MEN-SG is not toxic to rat platelets despite its redox cycling capacity and glutathione conjugation reaction of menadione could be regarded as a detoxification process.

Menadione에 의한 흰쥐 혈소판 세포독성에서 nitric oxide의 역할

승상애,김대병,윤여표,정진호 한국독성학회 1995 Toxicological Research Vol.11 No.2

Nitric oxide, a physiological transmitter, is reported to mediate cellular injury in various tissues. Its reactivity to free radical is believed to be one of the reasons for its involvement in cytotoxicity. Menadione, a representative quinone, is cytotoxic to several cell systems including isolated hepatocyte, endothelial cell and red blood cells. Its toxic mechanism is related to oxidative stress, mediated by toxic free radicals. Our previous studies demonstrated that menadione induced cell lysis and increase of oxygen consumption in platelets. It has been reported that platelets have nitric oxide producing enzyme, nitric oxide synthase. Thus, we have investigated to manifest the role of nitric oxide.in menadione-induced cytotoxicity in rat platelets. Menadione induced cytotoxicity in platelets was unaffected by $N^G$-nitro-arginine methyl ester (L-NAME), selective and competitive inhibitor of nitric oxide synthase. We also invesitgated the role of extracellular nitric oxide in menadione-induced cytotoxicity of platelets by addition with sodium nitroprusside (SNP). SNP did not affect platelet cytotoxicity by menadione. These results suggested that nitric oxide which was generated endogeneously or exogeneously might have a negligible role in menadione-induced cytotoxicity in rat platelets.

Menadione-induced Cytotoxicity in Rat Platelets: Absence of the Detoxifying Enzyme, Quinone Reductase

Kim, Kyung-Ah,Kim, Mee-Jeong,Ryu, Chung-Kyu,Chang, Moon-Jeong,Chung, Jin-Ho The Pharmaceutical Society of Korea 1995 Archives of Pharmacal Research Vol.18 No.4

The elevation of intracellular <TEx>$Ca^{2+}$ in various tissue through oxidative stress induced by menadione has been well documented. Increase of <TEx>$Ca^{2+}$ level inplatelets results in aggreaction of patelets. To test the hypothesis that menadione-induced <TEx>$Ca^{2+}$ elevations can play a role in platelet aggregation, we have studied the effect of menadione on aggragation of platelets isolated from female rats. Treatment with menadione to platelet rich plasma (PRP), which proved to be 60% as determined by aggregometry. however, exposure of PRP to menadione leads to a loss of cell viability, as measured by lactae dehydrogenase (LDH) leakage, suggesting that menadione might induce cell lysis rather than aggregation of platelets. Turbidty changes induced by menadione were unaffected by addition ofl dicoumarol, which is a quinone reducellular factions of patelets. These data, which indicate an absence of the QR detoxifying pathway, suggest that platelets may be more susceptible to menadione-induced cytotoxicity than certain other cell, as hepatocytes.

Apoptosis Induction by Menadione in Human Promyelocytic Leukemia HL-60 Cells

Duck-Jin Sa,Eun-Jee Lee,Byung-Sun Yoo 한국독성학회 2009 Toxicological Research Vol.25 No.3

Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration (1~50 μM) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 μM) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 μM of menadione, and very low at 100 μM of menadione. Taken together, our results showed that menadione induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.

Apoptosis Induction by Menadione in Human Promyelocytic Leukemia HL-60 Cells

Sa, Duck-Jin,Lee, Eun-Jee,Yoo, Byung-Sun Korean Society of ToxicologyKorea Environmental Mu 2009 Toxicological Research Vol.26 No.3

Cell death induced by menadione (vitamin K-3,2-methyl-1,4-naphthoquinone) has been investigated in human promyelocytic leukemia HL-60 cells. Menadione was found to induce both apoptosis and necrosis in HL-60 cells. Low concentration ($1{\sim}$50 ${\mu}$M) of menadione induced apoptotic cell death, which was demonstrated by typical DNA ladder patterns on agarose gel electrophoresis and flow cytometry analysis. In contrast, a high concentration of menadione (100 ${\mu}$M) induced necrotic cell death, which was demonstrated by DNA smear pattern in agarose gel electrophoresis. Necrotic cell death was accompanied with a great reduction of cell viability. Menadione activated caspase-3, as evidenced by both increased protease activity and proteolytic cleavage of 116 kDa poly(ADP-ribose) polymerase (PARP) into 85 kDa cleavage product. Caspase-3 activity was maximum at 50 ${\mu}$M of menadione, and very low at 100 ${\mu}$M of menadione. Taken together, our results showed that menadione induced mixed types of cell death, apoptosis at low concentrations and necrosis at high concentrations in HL-60 cells.

Menadione에 의한 3T3-L1 adipocytes의 사멸에 대한 프로폴리스의 보호효과

문준연(Jun-Yeon Moon),윤석환(Seok-Hwan Yoon),유병선(Byung-Sun Yoo) 한국양봉학회 2009 韓國養蜂學會誌 Vol.24 No.2

The study was conducted to investigate the protective effects of propolis against menadione-induced cell death of 3T3-L1 adipocytes. 3T3-L1 adipocytes were treated with menadione and cell viability was measured by MTT assay. The viability of 3T3-L1 adipocytes treated with 40μM menadione for 6h was significantly decreased to 41.4%, as compared to control. To investigate the protective effects of propolis against menadione treatment, 3T3-L1 adipocytes were treated with propolis 2h before 40μM menadione treatment, and cell viability was determined as MTT assay. The viability of 3T3-L1 adipocytes was significantly increased in cells pre-treated with 3㎍/㎖ propolis. The protective effects of propolis on 3T3-L1 adipocytes treated with menadione was high in cells treated with propolis 2h before menadione treatment, whereas not high in both cells treated with menadione 24h before and concomitantly. These findings show that propolis can protect menadione-induced cell death, through antioxidant effects.

Menadione Induced Apoptosis in MKN45 Cells via Down-regulation of Survivin

Min Ho Lee,Jeongyong Kim,Yoonjung Cho,Do Hyun Kim,Ji Yeong Yang,Hye Jin Kwon,Min Park,Hyun Jun Woo,Jong-Bae Kim,김종배 대한임상검사과학회 2019 대한임상검사과학회지(KJCLS) Vol.51 No.1

Menadione is known as an anti-tumor factor. Many studies have reported the potential anti-cancer role of menadione against a range of cancer cell lines. In this study, the anti-cancer effects of menadione and the underlying molecular signaling involved in apoptosis was investigated in gastric cancer cell lines. The menadione treatment decreased the cell viability of MKN45 gastric cancer cells. The decreased cell viability was attributed to the induction of apoptosis, which was confirmed by the results indicating the activation of caspase-3 and -7 and the cleavage of PARP in Western blotting. The upstream regulatory molecules involved in apoptosis were investigated further and it was discovered that menadione reduced the expression of survivin, an inhibitor of upstream apoptosis proteins. In addition, a transcription factor β–catenin, which is known to regulate survivin expression, was down-regulated by menadione. A previous report showed that menadione inhibited XIAP expression to induce apoptosis and induced G2/M cell cycle arrest in AGS cells. This study elucidated another inhibitory mechanism of menadione against gastric cancer cells in a different cell line. Although further studies will be needed, the inhibitory mechanism demonstrated in this study will help better understand the anti-cancer effects of menadione.

Menadione과 Plasma내의 Protein Thiol의 비효소적인 화학반응에 의한 활성산소 생성

정선화,이무열,이주영,장문정,정진호 한국독성학회 1997 Toxicological Research Vol.13 No.3

Quinones have been reported to undergo nonenzymatic reaction with thiols to generate reactive oxygens. It is therefore possible that the nonenzymatic reaction of quinones with thiols in plasma could lead to potentJared cellular toxicity or disease. When 1 mM menadione was added in plasma under pH 11.2, 7.4 and 5.0, the increase in oxygen consumption rate was the order of pH 11.2 > pH 7.4 > pH 5.0. In addition, oxygen consumption rates under plasma anticoagulated with trisodium citrate solution (pH 7.85) was significantly higher than those with acid-citrate-dextrose solution (pH 6.87). SOD and catalase reduced the rate of oxygen consumption induced by menadione in plasma. Taken together, these results suggest that the menadione-induced increased oxygen consumption was due to nonenzymatic reaction of menadione with thiols in the plasma. The presence of plasma has an additive effect on the increased oxygen consumption rates induced by the menadione treatments on our model tissue, platelets, as compared between washed platelet (WP) and platelet rich plasma (PRP). Cytotoxicity, as determined by LDH release, are well correlated with the oxygen consumption rates observed in each system and strongly suggest that menadione-induced cytotoxicity can be increased with the presence of blood plasma.

메나디온에 의한 혈소판 내 칼슘 변화측정시 형광 색소 사용의 문제점

정선화(Sun Hwa Chung),이무열(Moo Yeol Lee),이주영(Joo Young Lee),정승민(Seung Min Chung),정진호(Jin Ho Chung) 대한약학회 1997 약학회지 Vol.41 No.6

It has been reported that dose-dependent Ca2+ increase by menadione in platelets could be measured by fluorescent dye, quin-2. The problems will be described here relating to measuring Ca2+ in menadione-exposed platelets using fura-2 and fluo-3, widely used fluorescent indicators. Additions of menadione to fura-2 loaded platelets and their lysates resulted in marked reduction in fluorescence intensity at both 340nm (Ca2+-unbound form) excitation wavelengths. Fura-2 excitation spectra were overlapped with UV-visible absorption spectra of menadione, suggesting that light absorption by menadione itself could quench fluorescence generated by fura-2. Next approach was to use fluo-3 which has the higher wavelength (490nm) of excitation. Previous work demonstrated that treatment with probenecid to platelets was required to prevent fluo-3 dye leakage. However, probenecid itself was proven to be inadequate to measure the concentration of intracellular Ca2+ by reducing menadione-induced cytotoxicity in platelets. Our results suggest that it is not feasible to measure Ca2+ in platelets by using fura-2 and fluo-3 in the presence of probenecid, and cautions should be taken to measure changes of intracellular Ca2+ levels by fluorescent dyes following chemical exposure.