Down-Regulation of MITF, TRP-1, TRP-2, and Tyrosinase Expressions by Compounds Isolated from Pruni persicae Flos in Murine B16F10 Melanoma

이진영 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

Three single compounds were isolated by the fractionationfrom Pruni persicae Flos, and its chemical structure was analyzedusing 1H-, 13C-nuclear magnetic resonance, fast atom bombardment-Mass, and fourier transform infrared spectrometer analyses. Thecompounds were determined as quercetin 3-O-rhamnoside,kaempferol 3-O-β-neohesperidoside, and kaempferol 3-O-β-Dglucoside. To prove the whitening effect of these compounds,B16F10 melanoma was treated with quercetin-3-O-rhamnoside,kamferol-3-O-β-neohesperidoside or kaemferol-3-O-β-D-glucosideto confirm cell cytotoxicity using 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl-tetrazolium-bromide assay. As a result, cell cytotoxicitywas examined at more than 20% in 20 μg/mL of compoundsisolated from Pruni persicae Flos. Tyrosinase activity and melaninproduction were examined at 10 μg/mL of each compounds. Results showed quercetin-3-O-rhamnoside, kamferol-3-O-β-neohesperidosideor kaemferol-3-O-β-D-glucoside reduced tyrosinaseactivity in a dose-dependent manner. Moreover, quercetin-3-Orhamnosideshowed reduction of tyrosinase activity by 37.6% at10 μg/mL of quercetin-3-O-rhamnoside as well as the reductionof melanin contents by 58.8%. In addition, quercetin-3-Orhamnoside,kamferol-3-O-β-neohesperidoside, and kaemferol-3-O-β-D-glucoside reduced tyrosinase, tyrosinase-related proteins 1,and 2, microphthalmia associated transcription factor expressions,which were related to whitening effect, as well as both of mRNAand protein level of cyclic adenosine monophosphate as upstreamsignals. More interestingly, quercetin-3-O-rhamnoside stronglyreduced both mRNA and protein expressions than other compounds. Therefore, we determined the whitening effect from isolatedcompound fractionated from Pruni persicae Flos. Moreover, strongwhitening activity of quercetin-3-O-rhamnoside was examined,which suggested the potential application of this compound infunctional cosmetic development.

Down-Regulation of MITF, TRP-1, TRP-2, and Tyrosinase Expressions by Compounds Isolated from Pruni persicae Flos in Murine B16F10 Melanoma

Lee, Jin-Young The Korean Society for Applied Biological Chemistr 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

Three single compounds were isolated by the fractionation from Pruni persicae Flos, and its chemical structure was analyzed using $^1H$-, $^{13}C$-nuclear magnetic resonance, fast atom bombardment-Mass, and fourier transform infrared spectrometer analyses. The compounds were determined as quercetin 3-O-rhamnoside, kaempferol 3-O-${\beta}$-neohesperidoside, and kaempferol 3-O-${\beta}$-$\small{D}$-glucoside. To prove the whitening effect of these compounds, B16F10 melanoma was treated with quercetin-3-O-rhamnoside, kamferol-3-O-${\beta}$-neohesperidoside or kaemferol-3-O-${\beta}$-$\small{D}$-glucoside to confirm cell cytotoxicity using 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyl-tetrazolium-bromide assay. As a result, cell cytotoxicity was examined at more than 20% in $20{\mu}g/mL$ of compounds isolated from Pruni persicae Flos. Tyrosinase activity and melanin production were examined at $10{\mu}g/mL$ of each compounds. Results showed quercetin-3-O-rhamnoside, kamferol-3-O-${\beta}$-neohesperidoside or kaemferol-3-O-${\beta}$-$\small{D}$-glucoside reduced tyrosinase activity in a dose-dependent manner. Moreover, quercetin-3-Orhamnoside showed reduction of tyrosinase activity by 37.6% at $10{\mu}g/mL$ of quercetin-3-O-rhamnoside as well as the reduction of melanin contents by 58.8%. In addition, quercetin-3-Orhamnoside, kamferol-3-O-${\beta}$-neohesperidoside, and kaemferol-3-O-${\beta}$-$\small{D}$-glucoside reduced tyrosinase, tyrosinase-related proteins 1, and 2, microphthalmia associated transcription factor expressions, which were related to whitening effect, as well as both of mRNA and protein level of cyclic adenosine monophosphate as upstream signals. More interestingly, quercetin-3-O-rhamnoside strongly reduced both mRNA and protein expressions than other compounds. Therefore, we determined the whitening effect from isolated compound fractionated from Pruni persicae Flos. Moreover, strong whitening activity of quercetin-3-O-rhamnoside was examined, which suggested the potential application of this compound in functional cosmetic development.

Validation of an HPLC/UV-based method for Salicornia herbacea-derived isorhamnetin-3-O-glucoside and quercetin-3-O-glucoside quantification

( Jun Yeon Park ),( Leo Adrianne Paje ),( Ki Sung Kang ),( Sanghyun Lee ) 한국응용생명화학회 2021 Journal of Applied Biological Chemistry (J. Appl. Vol.64 No.3

Salicornia herbacea is a type of salt marsh plant that has been used in traditional medicine to treat several diseases. Isorhamnetin-3-O-glucoside (I3G) and quercetin-3-O-glucoside (Q3G) are major flavonoids in S. herbacea that are known to exert various pharmacological activities. Therefore, our study sought to validate and optimize an HPLC/UV-based analytical method for I3G and Q3G yield quantification, as well as to determine its limit of detection, limit of quantification, linearity, precision, and accuracy. Upon testing a concentration range of 31.5-1.9 μg/mL the results exhibited good linearity (r2 ≥0.9996 and r2 ≥0.9999 for I3G and Q3G, respectively), and the procedure was deemed precise (relative standard deviation of ≤3.19 and ≤3.85%, respectively), and accurate (102.6-105.0 and 92.9-95.2%, respectively). The results showed that our proposed method could be used for rapid I3G and Q3G evaluation in S. herbacea.

Flavonoids 및 그 배당체의 산화적 스트레스에 대한 신경교세포 보호 효과

김지현(Ji Hyun Kim),김현영(Hyun Young Kim),조은주(Eun Ju Cho) 한국생명과학회 2019 생명과학회지 Vol.29 No.12

뇌에서 과량의 reactive oxygen species (ROS) 생성에 의해 유발되는 산화적 스트레스는 알츠하이머 질환과 같은 신경퇴행성 질환의 원인으로 알려져 있다. 본 연구에서는 kaempferol, kaempferol-3-O-glucoside, quercetin, quercetin-3-β-D-glucoside와 같은 flavonoid와 그 배당체의 H₂O₂ 유도 산화적 스트레스에 대한 C6 신경교세포보호 효과를 확인하였다. H2O2만을 처리한 control군은 아무것도 처리하지 않은 normal군에 비해 세포 생존율 감소와 ROS 생성 증가를 통해 C6 신경교세포의 산화적 손상이 유도되었음을 확인하였다. 반면 4가지 flavonoid를 각각 처리한 군의 경우, H₂O₂를 처리한 control군에 비해 세포 생존율 증가와 ROS 생성 감소를 통해 산화적 손상 억제를 통한 신경교세포 보호 효과를 확인하였다. Flavonoid의 신경교세포 보호 작용 메커니즘을 규명하기 위해, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and interleukine (IL)-1β 등의 염증 관련 단백질 발현을 측정하였다. H₂O₂를 처리한 control군은 normal군에 비해 iNOS, COX-2, IL-1β 단백질 발현 증가와 IκB-α 발현 감소를 통해 신경교세포의 산화적 손상으로 인한 염증 반응을 확인하였다. 반면, 4가지 flavonoid를 각각 처리한 군의 경우 iNOS, COX-2, IL-1β 단백질 발현 감소와 IκB-α 발현 증가를 나타내어, 염증 반응 개선을 통한 신경교세포 보호 효과를 확인하였다. 특히, quercetin과 그 배당체인 quercetin-3-β-D-glucoside를 처리한 군은 kaempferol과 그 배당체인 kaempferol-3-O-glucoside를 처리한 군에 비해 우수한 신경교세포 보호 효과를 나타내었다. 본 연구는 4가지 flavonoid가 신경교세포에서 산화적 스트레스 억제를 통해 신경퇴행성 질환을 예방 및 치료할 수 있을 것으로 사료된다. Oxidative stress induced by the over-production of reactive oxygen species (ROS) in the brain is the most common cause of neurodegenerative diseases such as Alzheimer’s. In the present study, we investigated the protective effects of flavonoids and their glycosides, namely kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-β-D-glucoside, against H₂O₂-induced oxidative stress in the C6 glial cells. The H₂O₂-treated glial cells exhibited decreased cell viability and increased ROS production when compared with normal cells. However, cells treated with each of the four flavonoids/glycosides demonstrated significantly increased viability and suppressed ROS production when compared with the H₂O₂-treated control group. These results indicate that flavonoids/glycosides attenuate oxidative stress induced by H₂O₂ in C6 glial cells. To confirm the protective molecular mechanisms, we measured pro-inflammatory factors such as inducible nitric oxide synthase, cyclooxygenase-2, and interleukin-1β. H₂O₂ treatment was seen to elevate these factors and decrease IκB-α in the C6 glial cells, while the flavonoids/glycosides induced a down-regulation of the pro-inflammatory factors and increased IκB-α, indicating a neuroprotective effects through attenuation of the inflammation. In particular, quercetin and its glycoside showed a higher neuroprotective effect than the kaempferol treatments. These results suggest that these flavonoids and their glycosides could be promising therapeutic agents for neurodegenerative diseases via the attenuation of oxidative stress.

3-<i>O</i>-Glucosylation of quercetin enhances inhibitory effects on the adipocyte differentiation and lipogenesis

Lee, Chang Won,Seo, Jeong Yeon,Lee, Jisun,Choi, Ji Won,Cho, Sarang,Bae, Jae Youn,Sohng, Jae Kyung,Kim, Sung Oog,Kim, Jihoon,Park, Yong Il EDITIONS SCIENTIFIQUES ELSEVIER 2017 BIOMEDICINE AND PHARMACOTHERAPY Vol.95 No.-

<P><B>Abstract</B></P> <P>Glycosylation of natural flavonoids with various sugar moieties can affect their physicochemical and pharmacological properties. In this study, the plant flavonoids quercetin aglycon (Quer) and quercetin 3-<I>O</I>-glucoside (Q3G) were evaluated and compared for their potential anti-obesity effects. The Q3G dose-dependently reduced the TG contents and lipid accumulation in 3T3-L1 adipocyte cells, by 52% and 60% at 20μM, respectively, compared to differentiated control (100%), which were 1.6-fold and 2.4-fold higher reduction than Quer. The Q3G (20μM) also more significantly reduced the expression of adipogenic markers such as C/EBP-β, C/EBP-α, PPAR-γ, and aP2 than Quer, indicating that the Q3G suppresses both adipocyte differentiation and lipogenesis more effectively than Quer <I>in vitro</I>. Comparing to those in the high-fat diet (HFD) fed mice control group for 10 weeks, both the body and liver weights and the size of adipocytes in epididymal adipose tissues were significantly reduced in HFD mice fed with Q3G for another 6 weeks (30mg/kg body weight by oral administration), accompanied by the reductions of TG, total cholesterol, and HDL-cholesterol in serum. The Q3G also reduced the levels of the lipid metabolism-associated proteins, PPAR-γ, SREBP-1c, and FAS in the liver tissues. These results clearly demonstrated that Q3G exhibits a stronger anti-obesity effect than Quer and its anti-obesity effect is mediated via inhibition of adipocyte differentiation and lipogenesis, decreasing serum lipid levels by altering hepatic lipid metabolism, and reducing body weight gain. The results of this study suggest that the Q3G, but not Quer, can be a potent functional ingredient of beneficial health foods or a therapeutic agent to prevent or treat obesity.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Kaempferol, quercetin 및 그 배당체의 amyloid beta 유도 신경독성에 대한 C6 신경교세포 보호 효과

김지현,김현영,조은주 한국응용생명화학회 2019 Journal of Applied Biological Chemistry (J. Appl. Vol.62 No.4

Alzheimer’s disease (AD) is a common neurodegenerative disease. Oxidative stress by amyloid beta peptide (Aβ) of neuronal cell is the most cause of AD. In the present study, protective effects of several flavonoids such as kaempferol (K), kaempferol-3-O-glucoside (KG), quercetin (Q) and quercetin- 3-β-D-glucoside (QG) from Aβ25-35 were investigated using C6 glial cell. Treatment of Aβ25-35 to C6 glial cell showed decrease of cell viability, while treatment of flavonoids such as Q and QG increased cell viability. In addition, treatment of flavonoids declined reactive oxygen species (ROS) production compared with Aβ25-35- induced control. The ROS production was increased by treatment of Aβ25-35 to 133.39%, while KG and QG at concentration of 1 μM decreased ROS production to 107.44 and 113.10%, respectively. To study mechanisms of protective effect of these flavonoids against Aβ25-35, the protein expression related to inflammation under Aβ25-35-induced C6 glial cell was investigated. The results showed that C6 glial cell under Aβ25-35-induced oxidative stress up-regulated inflammation-related protein expressions. However, treatment of flavonoids led to reduction of protein expression such as inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1β. Especially, treatment of KG and QG decreased more effectively inflammation-related protein expression than its aglycones, K and Q. Therefore, the present results indicated that K, Q and its glycosides attenuated Aβ25-35-induced neuronal oxidative stress and inflammation. 알츠하이머 질환(Alzheimer’s disease)은 대표적인 신경퇴행성 질환이며, 뇌 내 amyloid beta (Aβ) 축적에 의한 산화적 스트레스 및 염증반응은 대표적인 AD의 원인으로 알려져 있다. 본 연구에서는 kaempferol (K), kaempferol-3-O-glucoside (KG), quercetin (Q) 및 quercetin-3-β-D-glucoside (QG)와 같은 4가지 flavonoids 의 C6 신경교세포에서 Aβ로 인한 신경독성으로부터의 보호 효능을 살펴보고자 하였다. C6 신경교세포에 Aβ를 처리하였을 때세포 생존율이 감소한 반면, 4가지 flavonoids 중에서 Q와 QG 의 처리 시 세포 생존율 증가를 통해 신경교세포 보호 효과를확인하였다. 또한, Aβ를 처리한 control군의 경우 reactive oxygen species (ROS) 생성을 유도한 반면, flavonoids의 처리 시 ROS 생성이 감소하였다. 특히 Aβ를 처리한 control군은 133.39%의ROS 생성을 나타내었으며, 1 μM의 KG와 QG를 각각 처리시107.44, 113.10%의 수치를 나타내어 ROS 생성 감소를 확인하였다. Flavonoids의 Aβ에 대한 신경교세포 보호 기전을 확인하기 위해 염증 관련 단백질 발현을 측정하였다. Aβ로 신경독성이 유도된 control군은 염증 관련 단백질 발현이 증가하였다. 그러나, flavonoids를 처리한 군의 경우 염증 매개 인자인 inducible nitric oxide synthase, cyclooxygenase-2 and interleukin-1β의 발현 감소를 확인하였다. 특히, KG와 QG를 처리한 군은aglycone 형태인 K와 Q를 처리한 군에 비해 효과적으로 염증매개 인자 발현을 감소시켰다. 본 연구는 flavonoids의 일종인K, Q와 그 배당체인 KG, QG의 Aβ로 신경독성이 유도된 신경교세포에서 산화적 스트레스 및 염증반응 조절을 통한 보호 효과를 나타냄을 알 수 있었으며, 이들 생리활성성분은 AD 예방및 치료 소재로써의 가능성이 있을 것으로 사료된다.

Kaempferol, quercetin 및 그 배당체들의 apoptosis 조절을 통한 신경세포 보호 효과

김지현(Ji Hyun Kim),이상현(Sanghyun Lee),조은주(Eun Ju Cho),김현영(Hyun Young Kim) 한국산학기술학회 2019 한국산학기술학회논문지 Vol.20 No.2

알츠하이머 질환은 대표적인 신경퇴행성 질환으로, 뇌 내에서 Aβ 단백질 축적은 알츠하이머 질환의 원인으로 알려져 있다. 본 연구에서는 amyloid beta (Aβ)로 손상을 유도한 SH-SY5Y 신경세포에서 kaempferol, kaempferol-3-O-glucoside, quercetin, quercetin-3-β-D-glucoside의 신경세포 보호 효과에 대해 검토하였다. SH-SY5Y 신경세포에 Aβ25-35 (25 μM)를 처리하였을 때, 처리하지 않은 normal군에 비해 세포생존율이 유의적으로 감소하였다. 반면, kaempferol, quercetin 및 그 배당체들을 각각 처리하였을 때 Aβ25-35만을 처리한 control군에 비해 유의적으로 세포생존율의 증가를 나타내었다. 또한, apoptosis에 관여하는 cleaved caspase9, Bcl-2-associated X protein (Bax) 단백질 발현을 측정한 결과, normal군에 비해 control군에서 유의적으로 cleaved caspase9 및 Bax 단백질 발현의 증가를 나타내어 Aβ 유도 신경세포 손상으로 인한 apoptosis가 유발됨을 확인하였으며, kaempferol, quercetin 및 그 배당체들의 처리 시 apoptosis 관련 단백질 발현이 감소함으로써 신경세포 보호 효능이 나타냄을 확인하였다. 이러한 결과는 kaempferol, quercetin 및 그 배당체들이 apoptosis 조절을 통해 신경세포 보호 효과를 나타내며, 신경세포 손상으로 인한 알츠하이머 질환을 예방하는 유용한 소재로써 사용 가능성이 있음을 보여준다. Alzheimer"s disease (AD) is a neurodegenerative disease caused by accumulation of amyloid beta (Aβ) in the brain. In the present study, we investigated the neuroprotective effects of four flavonoids such as kaempferol, kaempferol-3-O-glucoside, quercetin, and quercetin-3-β-D-glucoside against neuronal apoptosis induced by Aβ in SH-SY5Y neuronal cells. Treatment with Aβ decreased cell viability compared to the non-treated normal group. However, treatment with the four flavonoids increased cell viability in SH-SY5Y cells treated with Aβ. In addition, we measured the expression of apoptosis-related proteins such as Bcl-2-associated X protein (Bax) and cleaved caspase-9. Treatment with the four flavonoids down-regulated Bax and cleaved caspase-9 in Aβ-treated SH-SY5Y neuronal cells. Overall, the results of the present study demonstrated the neuroprotective effect of flavonoids by anti-apoptotic activity in Aβ-induced SH-SY5Y neuronal cells. These results suggest that these four flavonoids would be useful therapeutic and prevention agents for AD.

대극 지상부의 페놀성 화합물(제2보)

안병태,강삼식,Ahn, Byung-Tae,Kang, Sam-Sik 한국생약학회 1996 생약학회지 Vol.27 No.2

We have previously reported the isolation of seven hydrolyzable tannins and nine flavonoids from Euphorbia pekinensis Ruprecht. Further investigation about the same plant has led to the isolation of kaempferol, quercetin, kaempferol $3-O-(2'-O-galloyl-{\beta}-_D-glucoside)$, quercetin 3-O-(2'-O-galloylrutinoside), ellagic acid and acetonyl geraniin. These compounds were isolated from this plant for the first time.

Euphorbia ebracteolata에 대한 생약학적 연구(I) -Euphorbia ebracteolata의 Flavonoid 성분-

이상철,안병태,박웅양,이승호,노재섭,이경순,유응걸,Lee, Sang-Cheol,Ahn, Beung-Tae,Park, Woong-Yang,Lee, Seung-Ho,Ro, Jai-Seup,Lee, Kyong-Soon,Ryu, Eung-Kul 한국생약학회 1992 생약학회지 Vol.23 No.3

Four flavonoids were isolated from the aerial parts of Euphorbia ebracteolata. On the basis of chemical and spectroscopic evidence, the structures of these compounds were established as $quercetin-3-O-{\beta}-D-glucoside$(isoquercitrin)(I), quercetin-3-O-rutinoside(rutin)(II), kaempferol-3-O-rutinoside(III) and $quercetin-3-O-(2'-O-galloyl)-{\beta}-D-glucoside$(IV) which was the main flavonoidal component in this plant. The isolation of flavonoids from E. ebracteolata is the first example.

Pharmacokinetic properties of monotropein, kaempferol 3-O-glucoside and quercetin 4"-O-glucoside after oral and intravenous gavage of MOTILIPERM in rats

Bo Ram Choi,Keshab Kumar Karna,Yu Seob Shin,Sung Won Lee,Chul Young Kim,Hye Kyung Kim,Jong Kwan Park 한국실험동물학회 2019 한국실험동물학회 학술발표대회 논문집 Vol.2019 No.7