Betulinic Acid Stimulates Glucose Uptake through the Activation of PI3K and AMPK in 3T3-L1 Adipocytes

Jung Kyung Lee(이정경),Jae Eun Park(박재은),Ji Sook Han(한지숙) 한국생명과학회 2022 생명과학회지 Vol.32 No.10

제 2형 당뇨병은 고혈당을 특징으로 하는 만성 대사성 질환으로 인슐린 민감성및 저항성을 개선하여 세포속으로 포도당 흡수를 촉진시킴으로서 완화될 수 있다. 본 연구는 triterpenoid 화합물인 betulinic acid가 3T3-L1 지방세포에서 인슐린 신호전달체계를 개선하여 포도당 흡수를 촉진시키는지를 조사하고 그 작용기전을 규명하였다. Betulinic acid는 3T3-L1 지방세포에서 농도의존적으로 포도당 흡수를 유의하게 증가시켰으며, 이는 PM-GLUT4의 발현 증가와 관련이 있음을 관찰하였다. Betulinic acid는 인슐린 신호전달 경로에서 PI3K의 활성화 및 IRS-1tyr, Akt의 인산화를 대조군에 비해 유의하게 증가시켰다. 또한 AMPK의 활성화를 나타내는 pAMPK와 AMPK 하위인자인 pACC의 수준을 유의하게 증가시켰다. Betulinic acid에 의한 PI3K 및 AMPK 경로의 활성화를 증명하기 위해, PI3K 억제제(Wortmannin)와 AMPK의 억제제(Compound C)를 사용하여 이들 처리에 의한 포도당 흡수능과 PM-GLUT4의 발현을 측정한 결과 이들의 발현이 유의하게 저해되었다. 본 연구에서 betulinic acid는 3T3-L1 지방세포에서 PI3K 및 AMPK 경로의 활성화를 통해 세포막으로 포도당 수송체인 GLUT4 전위를 촉진시키고 포도당 흡수를 증가시킬 수 있음을 나타내었다. 이러한 결과는 betulinic acid가 인슐린 민감성을 증진시키고 고혈당을 완화하는데 도움이 될 수 있음을 시사한다. Hyperglycemia in type 2 diabetes can be alleviated by promoting cellular glucose uptake. Betulinic acid (3β,-3-hydroxy-lup-20(29)-en-28-oic acid) is a pentacyclic lupane-type triterpenoid compound. Although there have been studies on the antidiabetic activity of betulinic acid, studies on cellular glucose uptake are lacking. We investigated the effects of betulinic acid on glucose uptake and its mechanism of action in 3T3-L1 adipocytes. Betulinic acid significantly stimulated glucose uptake in 3T3-L1 adipocytes by increasing the phosphorylation of the insulin receptor substrate 1-tyrosine (IRS-1tyr) in the insulin signaling pathway, which in turn stimulated the activation of phosphoinositide 3-kinase (PI3K) and the phosphorylation of protein kinase B (Akt). The activation of PI3K and Akt by betulinic acid translocated glucose transporter 4 to the plasma membrane (PM-GLUT4), thereby increasing the expression of PM-GLUT4 and thus stimulating cellular glucose uptake. Betulinic acid also significantly increased the phosphorylation/activation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase. The activation of PI3K and AMPK by betulinic acid was confirmed using the PI3K inhibitor wortmannin and the AMPK inhibitor compound C. The increase in glucose uptake induced by betulinic acid was significantly decreased by wortmannin and compound C in the 3T3-L1 adipocytes. These results suggest that betulinic acid stimulates glucose uptake by activating PI3K and AMPK in 3T3-L1 adipocytes.

Betulinic acid 등의 세포독성 조절 연구

염영나,조현영,김현석,황명실,윤은경,이효민,김승희,양지선,양기화 식품의약품안전청 2001 식품의약품안전청 연보 Vol.5 No.-

과일이나 식물체 등애 천연적으로 존재하면서 돌연변이억제나 항산화효과 등, 암이나 성인병 등에 대한 예방효과를 보이는 것으로 알려진 phytocherilical을 중심으로 aminopept건ase 활성 억제 능력을 조사하여 암전툴과정과 혈관신생과정에서 발된이 증가하는 것으로 알려진 arnjnoepeptidase N APh억제제를 스크리닝하고 암전이 예방이나 암치료에 활용하고자 본 연구를 수행하게 되었다 Leucine andnopeptidase에 대한 식물유래성분(betulinic acid, curcvmin, indomethacin, ellagic acid quercetin, nocodazole, urso:ic acid, resveratrol, h)'pericin, caffeic acid)의 찰성억제능력을 조사하여 그중 촬성억제능력이 탁월한 curcundn, indomethacin, betulinlc 3c펀의 APN enzyme netics를 수헝 하였다. Curcumin의 andnopeptidase 띠 활성을 억제하는 농도와 혈관신생의 억제 농도가 일치하고 APN 단클론항체 처리한 HT1080 세포의 유세포측정결과 curcumin·라 APN 단클론항체가 경정적으로 APN예 결합됨을 확인함에 따라 curcumin이 암전이를 억제하는 기전이 APN 억제에 기인한 것으로 추정되는 결과이다. 한편 Betulinic ac,김는 APN 활성에는 영향을 미치지 못하고, 미토콘드리아의 막전 위를 떤화시키고 caspase근 활성을 증가시켜 apoptosis를 유발하였다. 포한 betulinic acid로 tlibe formation assay와 」'n ufuo CAM assay를 수행한 결과 혈관신생과정을 억제하는 것이 확인되었다 따라저 Betulinic 3fid와 curcumin은 서로 작용기전은 다르지만, 암전이 떼방이나 암치료에 활용가치가 높은 약물로 인식되었다. This study is to evaluate phytochemicals that have inhibitory activity on aminopeptidase N (APN) which is transmembrane metalloprotease to play functional role in matrix degradation and invasion by tumor cells. At first, we did screening of APN inhibitors among the phytochemicals with chemopreventive effect including betulinic acid, curcumin, indomethacin, ellagic acid, quercetin, nocadazole, ursolic acid, reveratrol, hypericin, and caffeic acid etc. by the fluorometric assay using L-alanine 4-methylcoumaryl-7-amide(Ala-AMC) as a substrate. Curcumin has inhibitory effect on APN activity as well as angiogenesis in a same condition(IC_(50) value=about 10μM). Curcumin on APN activity works as a noncompeptitive ingibitor. FCM analysis shows the competitive binding of curcumin and monoclonal antibody of APN to HT1080 cell. This results means that curcumin blocks the angiogenesis by the mechanism of APN inhibition. Betulinic acid which is working as a inhibitor of Leu aminopeptidase, dose not inhibit the aminopeptidase N activitiy. But betulinic acid blocks angiogenesis from the results of tube formation assay and in vivo CAM assay. The angiogenesis blocking mechanism of betulinic acid is supposed to induce apoptosis. These results suggest that curcumin and betulinic acid can be used as angiogenesis blocker and chemopreventive agent of cancer despite of different mechanism.

호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 Betulinic Acid의 효과

김훈성,최윤석,이준혁,박나경,박창휘,이영하,김귀옥,송시연,배창훈,이승호,김용운,김용대 대한이비인후과학회 2014 대한이비인후과학회지 두경부외과학 Vol.57 No.8

Background and Objectives MUC5AC and MUC5B are representative secretory mucin genes in the human airway, whose expressions are increased by a variety of inflammatory mediators. Betulinic acid, a naturally occurring pentacyclic triterpenoid, is known to have an anti-inflammatory property. However, the effects of betulinic acid on mucin secretion of airway epithelial cells still have not been reported. Therefore, in this study, the effect of betulinic acid on inflammatory mediators-induced MUC5AC and MUC5B expressions was investigated in human airway epithelial cells. Materials and Method In the mucin-producing human NCI-H292 airway epithelial cells, the effects of betulinic acid on interleukin-1β (IL-1β)-, lipopolysaccharide (LPS)-, and phorbol myristate acetate (PMA)-induced MUC5AC and MUC5B expressions were analyzed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results Betulinic acid attenuated IL-1β-, LPS-, and PMA-induced MUC5B mRNA and glycoprotein expression in NCI-H292 cells. On the other hand, betulinic acid did not attenuate IL-1β-, and LPS-, but induced PMA-induced MUC5AC mRNA and glycoprotein expressions in NCI-H292 cells. Conclusion These results suggest that betulinic acid attenuates IL-1β-, LPS-, and PMA-induced MUC5B expression in the airway epithelial cells. Therefore, betulinic acid may modulate a control of mucus-hypersecretion in airway inflammatory diseases. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(8):526-32

호흡기 상피세포에서 MUC5AC와 MUC5B 발현에 대한 Betulinic Acid의 효과

김훈성 ( Hoon Sung Kim ),최윤석 ( Yoon Seok Choi ),이준혁 ( Jun Hyeok Lee ),박나경 ( Na Kyung Park ),박창휘 ( Chang Hwi Park ),이영하 ( Young Ha Lee ),김귀옥 ( Gui Ok Kim ),송시연 ( Si Young Song ),배창훈 ( Chang Hoon Bae ),이승호 영남대학교 약품개발연구소 2014 영남대학교 약품개발연구소 연구업적집 Vol.24 No.0

Background and Objectives MUC5AC and MUC5B are representative secretory mucin genes in the human airway. MUC5AC and MUC5B expression are increased by a variety of inflammatory mediators. Betulinic acid, a naturally occurring pentacyclic triterpenoid, is known to have an anti-inflammatory property. However, the effects of betulinic acid on mucin secretion of airway epithelial cells still have not been reported. Therefore, in this study, the effect of betulinic acid on inflammatory mediators-induced MUC5AC and MUC5B expression was investigated in human airway epithelial cells. Materials and Method In the mucin-producing human NCI-H292 airway epithelial cells, the effects of betulinic acid on interleukin-1β (IL-1β)-, lipopolysaccharide (LPS)-, and phorbol myristate acetate (PMA)-induced MUC5AC and MUC5B expression were analyzed by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Results Betulinic acid attenuated IL-1β-, LPS- and PMA-induced MUC5B mRNA and glycoprotein expression in NCI-H292 cells. Betulinic acid did not attenuate IL-1β-, LPS- and PMA-induced MUC5AC mRNA and glycoprotein expression in NCI-H292 cells. Conclusion These results suggest that betulinic acid attenuates IL-1β-, LPS- and PMA-induced MUC5B expression in airway epithelial cells. Therefore, betulinic acid may modulate a control of mucus-hypersecretion in airway inflammatory disease. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(8):526-32

Betulinic Acid Inhibits LPS-Induced MMP-9 Expression by Suppressing NF-kB Activation in BV2 Microglial Cells

( Sung Soo Kim ),( Jae Won Lee ),( Yong Joon Choi ),( Sue Young Lee ),( Sang Soo Kang ),( Nam Ho Kim ),( Yong Soo Kwon ),( Hee Jae Lee ),( Wan Joo Chun ) 한국응용약물학회 2011 Biomolecules & Therapeutics(구 응용약물학회지) Vol.19 No.4

Aberrant activation of microglia has been reported to cause neuronal damages by releasing a variety of pro-infl ammatory cytokines. Besides where microglia become active, damages have been also observed in remote places, which is considered due to the migration of activated microglia. Therefore, an agent that could suppress abnormal activation of microglia and their subsequent migration might be valuable in activated microglia-related brain pathologies. The objective of the present study was to evaluate anti-infl ammatory effects of betulinic acid on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Pretreatment of betulinic acid signifi cantly attenuated LPS-induced NO production and protein expression of iNOS. Betulinic acid also signifi cantly suppressed LPS-induced release and expression of cytokines such as TNF-α and IL-1β. Furthermore, betulinic acid signifi cantly suppressed LPS-induced MMP-9 expression, which has been suggested to play an important role in the migration of activated microglia. In order to understand the possible mechanism by which betulinic acid suppresses LPS-induced cytokine production and migration of microglia, the role of NF-kB, a major pro-infl ammatory transcription factor, was examined. Betulinic acid signifi - cantly suppressed LPS-induced degradation of IKB, which retains NF-kB in the cytoplasm. Therefore, nuclear translocation of NF-kB upon LPS stimulation was signifi cantly suppressed with betulinic acid. Taken together, the present study for the fi rst time demonstrates that betulinic acid possesses anti-infl ammatory activity through the suppression of nuclear translocation of NF-kB in BV2 microglial cells.

HL-60 세포에서 $TNF-{\alpha}$에 의한 MCP-1 발현에 미치는 Betulinic Acid의 효과

김경찬,이추희,Kim, Kyung-Chan,Lee, Chu-Hee 대한약학회 2008 약학회지 Vol.52 No.1

Betulinic acid, a naturally occurring pentacyclic triterpenoid, is found in abundance in the outer bark of white birch (Betula alba). In this study, we investigated if betulinic acid affects cytokine expression from activated macrophage cells. ELISA result showed that stimulation of HL-60 cells with proinflammatory cytokine such as $TNF-{\alpha}$ resulted in MCP-1 release into culture medium. In addition, transcriptional upregulation of MCP-1 in response to $TNF-{\alpha}$ was observed by RT-PCR analysis. However, incubation of HL-60 cells with betulinic acid prior to $TNF-{\alpha}$ treatment abrogated MCP-1 expression in transcription and translational level. Consistent with a number of studies which reported requirement of ERK activation for $TNF-{\alpha}$ expression, Western blot analysis showed that $TNF-{\alpha}-induced$ ERK activation was suppressed by pretreatment of HL-60 cells with betulinic acid. Taken together, our data indicate that betulinic acid exerts its anti-inflammatory effect through inhibition of $TNF-{\alpha}-induced$ ERK activation which is required for the subsequent MCP-1 release.

플라타너스(Platanus orientalis L.)의 계절별 Betulinic acid 함량 및 생리활성

김영균,고영남 國民大學校 山林科學硏究所 2001 山林科學 Vol.13 No.-

최근 폐암, 난소암, 흑색종 등 각종 악성종양 및 후천성면역결핍증(AIDS) 에 대하여 우수한 치료효과를 가지고 있다는 연구결과가 빈번하게 발표되고 있는 triterpene화합물 betulinic acid를 계절별로 플라타너스를 채취하고 MeOH 추출물로부터 분리하였다. 순수하게 정제된 betulinic acid의 함량을 비교하였고, 생물학적 활성을 검정하였다. A lupane type triterpenoid, betulinic acid has been known to have lung cancer, ovarian cancer, human melanoma and other tumor growth inhibitory activities and also anti-HIV activity. In this research, we collected P. orientalis in every seasons and observed seasonal changes of betulinic acid in the species. Cytotoxic activity of betulinic acid is also shown in this report.

대추나무 껍질 유래 Betulinic acid의 in vitro rotavirus 감염억제 효과

이경호,이기형,조좌형 한국생약학회 2008 생약학회지 Vol.39 No.2

This study was conducted to evaluate the inhibitory effects of betulinic acid isolated from Ziziphus jujuba on various human rotavirus, such as KU, S2 and YO. The results obtained are summarized as follows: At the concentration of betulinic acid 0.1/2%, all human rotavirus is showed a maximum effect on their growth even though it’s evaluated on the in vitro test. The each inhibitory rate of MA-104 cells infected by human rotavirus KU, S2 and YO was 62.1%, 59.7% and 65.2%, respectively, at the concentration of 0.1/2%. The anti-virus activity of betulinic acid was showed as the dose-dependent manner at the used dosages except at the dose of 0.1%.

국산 건대추의 외형 등급별 품질특성 연구

방미희(Mi Hui Bang),유혜영(Hye Young Yu),배봉석(Bong Seok Bae),박철수(Chol Su Park),한민우(Min Woo Han) 한국약용작물학회 2020 한국약용작물학회지 Vol.28 No.6

Background: The purpose of this study was to analyze the applicability of dried jujube (Zizypui Fructus) distributed by size as a functional food material and herbal medicine by evaluating its physicochemical characteristics and betulinic acid content. Methods and Results: It was shown that the carbohydrate content of in jujube fruit significantly increased with fruit size; 81.42% for large, 79.83% for medium, and 76.39% for small. Similarly, the dilute ethanol extract content of each group was 72.48% (large), 69.56% (medium), and 64.16% (small). A free sugar analysis indicated that glucose, fructose and sucrose were found in quantitieds proportional to jujube fruit size, the total free sugar values were 68.85%, 63.93%, and 57.37% for large, medium, and small fruit sizes respectively. The betulinic acid content for large, medium, and small fruit was 0.50 ㎎/g, 0.54 ㎎/g, and 0.58 ㎎/g respectively, indicating that smaller jujube contained a higher amount of betulinic acid. Conclusions: It is considered that betulinic acid content could be used as a criteria for jujube fruit quality control.

루판 유도체의 합성 및 세포독성 -I

유영제(Young Jae You),김용(Yong Kim),안병준(Byung Zun Ahn) 대한약학회 2002 약학회지 Vol.46 No.5

To observe the structure-activity relationship of lupane derivatives both on cytotoxic and aftiangiogenic activity twelve lupane derivatives were prepared; their antiangiogenic and cytotoxic activities were evaluated. Among them, four compounds were more cytotoxic than betulinic acid. Carboxylic acid at C28 seemed to be essential for cytotoxic activity. But, a selective cytotoxicity toward SK-MEL-2 was not observed. As for antiangiogenic activity none of the compounds except lupeol showed antiangiogenic activity at 30 μg/mL.