구리이온(II)이 존재할 때 Salvianolic acid B에 의한 DNA 절단

이평재,문철,최윤선,손현규,Lee, Pyeongjae,Moon, Cheol,Choi, Yoon Seon,Son, Hyun Kyu 대한임상검사과학회 2018 대한임상검사과학회지(KJCLS) Vol.50 No.2

단삼의 성분인 salvianolic acid B는 다양한 생리활성이 알려져 있다. 특히 항산화 효과는 간세포, 신경세포를 포함한 다양한 세포유형에서 보호효과가 있다고 보고되었다. 하지만 ferulic acid와 같이 항산화제로 여겨지는 몇몇 페놀성 물질은 특정 전이 금속이 있으면 산화작용을 하며 이것이 항암 효능을 설명하기도 한다. 본 실험에서 salvianolic acid B가 $Cu^{2+}$ 환경에서 산화작용을 하는지 알아보았다. salvianolic acid B와 $Cu^{2+}$를 동시 처리하면 supercoilded 형태의 DNA가 open circular 혹은 linear 형태로 바뀌었으나 salvianolic acid B 혹은 $Cu^{2+}$를 단독 처리 했을 때는 그렇지 않았다. $Cu^+$에만 특정적인 킬레이터 neocuproine을 이용하여 salvianolic acid B가 $Cu^{2+}$를 $Cu^+$로 환원시킴을 알았으며 $H_2O_2$를 물과 산소로 분해하는 catalase를 처리하면 DNA 분해가 일어나지 않았다. 활성산소종 중 하나인 $H_2O_2$는 생체분자 특히 DNA를 공격하여 정상기능을 수행하지 못하게 한다. 정리하면 salvianolic acid B에 의한 $Cu^{2+}$의 환원은 $H_2O_2$를 생성하며 $H_2O_2$는 DNA 분해를 일으킨다. 이런 결과는 salvianolic aicd B의 항암효과가 salvianolic acid의 $H_2O_2$ 생성 때문일 수 있다는 작은 단서를 줄 수 있으며 이는 좀 더 실험이 이뤄져야 한다. Salvianolic acid B, which is a compound in the Salvia miltiorrhiza, has diverse biological activities, In particular, the antioxidative effects were reported to be involved in the protection of hepatocytes, neurons, and various cell types. On the other hand, some phenolic compounds, such as ferulic acid, which is regarded as an antioxidant, plays a pro-oxidative role in the specific transitional metal environment, which could explain the anticancer effect. This study examined the pro-oxidative effects of salvianolic acid B in the presence of $Cu^{2+}$. Treatment with both salvianolic acid B and $Cu^{2+}$ induced the transition of supercoiled DNA to the open circular or linear form but not in the sole salvianolic acid B or $Cu^{2+}$ treatments. Salvianolic acid B reduced the $Cu^{2+}$ to $Cu^+$ using neocuproine, a $Cu^+$ specific chelator. In addition, catalase, an enzyme that breaks down the $H_2O_2$ to water and molecular oxygen, inhibited the DNA breakage. $H_2O_2$, a reactive oxygen species, has detrimental effects on biological molecules, particularly DNA. Overall, the reduction of $Cu^{2+}$ by salvianolic acid B could lead to the production of $H_2O_2$ followed by DNA breakage. These results suggest that the pro-oxidative effects could be the one of the anti-cancer mechanisms of salvianolic acid B, which remains to be explained.

연도별 국내산 품종과 중국산 단삼 품종의 약리활성물질 함량 및 변이

신예림,류병렬,강민지,김민준,정진태,한종원,이기욱,임정대,임정대 한국약용작물학회 2024 한국약용작물학회지 Vol.32 No.2

Background: This study was conducted to determine color parameters, and identify pharmaceutical constituents and phytochemical characteristics of Korean cultivars (Hongdan, Dasan, and Kosan) of Salvia miltiorrhiza root (Danshen) by crop years compared with those of the Chinese cultivar [“Zhongdanyaozhi No. 1 (ZD1)”]. Methods and Results: Redness (a*), yellowness (b*), extract yield, salvianolic acid B (Sal B), and tanshinone ⅡA (TanⅡA) were generally higer in Korean cultivars than in ZD1, implying effective quality control with Korean cultivars in Danshen. Sal B and TanⅡA content were found to be the highest in Korean cultivar ‘Hongdan’ (3-fold higher than ‘ZD1’) in the 2020 crop year. Sal B was chosen as a major markers to assess Korean Danshen quality. However, Sal B detection was apparently incomplete and insufficient for quality control, because Sal B degradation was crop years dependent, Sal B was degraded to some small molecular phenolic acids, such as caffeic acid, protocatechuic aldehyde, lithospermic acid, and danshensu, which were elucidated by liquid chromatography. Analysis of Pearson’s correlation heatmap revealed negative correlation between Sal B and small phenolic acids. The Sal B content of Danshen was correlated to the redness of color parameter, the total terpenoid content, TanⅡA content, suggesting that it would be reasonable to simultaneously detect these characteristics for Danshen quality control. Conclusions: This study provides constructive and meaningful references for the qualitative and quantitative analyses of S. miltiorrhiza root, which can help establish better quality standards.

Bioavailability of Salvianolic Acid B and Effect on Blood Viscosities after Oral Administration of Salvianolic Acids in Beagle Dogs

Dong-Yan Gao,Li-Mei Han,Li-Hong Zhang,Xiao-Ling Fang,Jian-Xin Wang 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.5

Salvianolic acid B (SalB) is an active component isolated from Chinese herbal medicine Salvia miltiorrhiza. The aim of this study was to investigate the extent of absolute oral bioavailability (F) of SalB in beagle dogs and the effect on blood viscosity after intravenous and oral administration of Salvianolic acids (SAs). A gradient elution HPLC method was developed and validated to determine the concentration of SalB and its three possible metabolites in plasma. After SAs (180 mg/kg, p.o.; 9 mg/kg, i.v.) were given, the AUCs of SalB were 1680 ± 670 and 7840 ± 1140 ng/mL·h, respectively. The F of SalB in dogs was calculated to be only 1.07 ± 0.43%. The blood viscosity was remarkably decreased after a single intravenous injection of SAs (9 mg/kg). However, no significant change of blood viscosity was observed after a single oral administration of SAs (180 mg/kg). The results suggested that the F of SalB was extremely low and single oral administrated SAs had no effect on ameliorating blood viscosity in beagle dogs.

Salvianolic acid B가 고강도 운동부하에 의한 흰쥐 골격근과 뇌조직의 Oxidative Stress에 미치는 영향

이현준 ( Hyun-joon Lee ),강성한 ( Sung-han Kang ),권수현 ( Su-hyeon Kweon ),김대경 ( Dae-kyung Kim ),김지호 ( Jeeho Kim ),문지홍 ( Ji-hong Moon ),신정원 ( Jung-won Shin ),이종수 ( Jong-soo Lee ),손낙원 ( Nak-won Sohn ) 대한본초학회 2016 大韓本草學會誌 Vol.31 No.5

Objectives : Salvianolic acid B (SAB) is an active ingredient in Salvia miltiorrhiza frequently used for cardiovascular and cerebrovascular diseases. The present study investigated the antioxidant effects of SAB on the skeletal muscle and the brain tissue of rats following exhaustive exercise. Methods : The rats were treated with oral administration of SAB (30 mg/kg) daily for 5 days prior to the exhaustive exercise. The exhaustive exercise was performed as swimming for 150 min with 5% body weight attached to the tail on the 5th day. The antioxidant effects of SAB was evaluated by measuring the superoxide generation in the gastrocnemius and the 4-HNE expression in the hippocampal tissue. In addition, c-Fos-expressing cells in the brain tissue was observed using immunohistochemistry. Results : Histological features and muscle fiber type composition were not different between the SAB group and the exhaustive exercise group. SAB significantly reduced the upregulation of superoxide generation in the muscle tissue. SAB significantly reduced the increase of c-Fos-expressing cells in the cerebral cortex, paraventricular thalamic nucleus, dorsomedial hypothalamic nucleus, the CA1, CA3, and DG regions of hippocampus. SAB significantly reduced the upregulation of 4-HNE expression in the CA1 and DG regions of hippocampus caused by the exhaustive exercise. Conclusions : The results suggest that SAB exerts antioxidative effect against oxidative stress in the skeletal muscle and the brain tissue following exhaustive exercise, while SAB may has an anti-stress effect on stress responses in the brain.

Large Scale Isolation and Purification of Salvianolic Acid B in High Purity from Roots of Dansham (Salvia miltiorrhiza Bunge)

Hyoung Jae Lee,Ki Hoon Lee,박근형,문제학 한국식품과학회 2010 Food Science and Biotechnology Vol.19 No.2

Salvianolic acid B (Sal B) is the most abundant representative compound in roots of dansham (Salvia miltiorrhiza) which is beneficial in treatment of cardiovascular disease. The present study was performed to establish a large scale isolation method of Sal B in high purity from dansham roots. Water suspension of dansham roots’ (100 g d.w.) methanol extract (13.16 g) was partitioned with chloroform, and the water layer was adjusted to pH 3.0 by triflouroacetic acid, because the Sal B partition coefficient in acidic condition was predominately higher than that in neutral condition, and then continuously extracted with ethyl acetate (EtOAc). The EtOAc-soluble acidic fraction was subjected to Sephadex LH-20 column chromatography. The fractions (Ve/Vt 1.75-2.71, 1.82 g) containing Sal B were finally purified by preparative high performance liquid chromatography using an octadecylsilane column. The isolated Sal B fraction (1.08 g) represented a 75.0%recovery, with a purity exceeding 99.2%.

Anti-adipogenic Effect and Mechanism in 3T3-L1 Preadipocyte Differentiation by Salvianolic Acid B

권효신,장병철 계명대학교 의과대학 2022 계명의대학술지 Vol.41 No.2

Salvianolic acid B (Sal B) is one of the most active hydrophilic compounds extracted from Salvia miltiorrhiza root. Previous in vitro and in vivo studies demonstrate the ability of Sal B to modulate adipocyte differentiation. However, the lipid-modulating effect and mechanism of Sal B in adipocytes remain controversial. Here we investigated the regulatory effect and mode of action of Sal B on lipid accumulation in 3T3-L1 preadipocyte differentiation. Lipid droplet (LD) accumulation and triglyceride (TG) content during 3T3-L1 preadipocyte differentiation were measured by Oil Red O staining and AdipoRed assay. The growth inhibition during 3T3-L1 preadipocyte differentiation was measured by cell count analysis. Western blotting and real-time qPCR analysis were utilized to determine the protein and mRNA expression in the preadipocyte differentiation. Notably, in 3T3-L1 preadipocyte differentiation, treatment with Sal B at 100 M led to a marked decrease in LD accumulation and TG content without influencing cell growth. Sal B treatment (100 M) further reduced the expression and phosphorylation levels of adipogenic transcription factors, including CCAAT/enhancer-binding protein- (C/EBP-), peroxisome proliferator-activated receptor-gamma (PPAR)-, and signal transducer and activator of transcription (STAT)-3/5. Treatment with Sal B (100 M) also reduced the expression and phosphorylation levels of fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC), two lipogenic enzymes and perilipin A, an LD-binding and stabilizing protein. These results collectively demonstrate that Sal B at 100 M strongly inhibits lipid accumulation in 3T3-L1 preadipocyte differentiation, mediated through regulation of the expression and phosphorylation levels of C/EBP-, PPAR-, STAT-3/5, FAS, ACC, and perilipin.

Salvianolic acid B가 자외선B 조사에 의한 plasmid DNA 절단에 미치는 효과

이평재 대한미용학회 2019 대한미용학회지 Vol.15 No.4

Ultraviolet B (UVB), which is 280-320 nm wavelength light, can induce modification of base and (or) DNA breakage by direct or indirect pathways. Generation of hydroxyl radical is one of the mechanisms by which UVB imparts detrimental effect on DNA. Salvianolic acid B (SalB) is one of the components in Salvia miltiorrhiza and has been reported to have antioxidant, anti-inflammation and anti-wrinkle effects. In the present study, SalB was found to protect the DNA breakage and reduce the 2-deoxy-d-ribose degradation under UVB radiation with H2O2. However, SalB increased the DNA breakage and degradation of 2-deoxy-d-ribose in the presence of Cu2+ ion. These results implied that SalB might have had anti-oxidant or pro-oxidant effect depending on the micro-environment. The current study was carried out with direct UVB radiation to DNA. The roles of SalB in the cell or tissue level still remain to be investigated.

고강도 운동부하에 의한 흰쥐의 심장근육 손상에 대한 Salvianolic Acid B의 효능에 대한 연구

임웅진,이종수 한방비만학회 2017 한방비만학회지 Vol.17 No.2

Objectives: This study examined the effects of salvianolic acid B (SAB) on exhaustive exerciseinduced cardiac muscle damage to rats. Methods: The study was carried out with 12-week-old young adult male Sprague-Dawley rats. Thirty-six rats were divided into 3 groups; normal (n=12), exhaustive exercise group (ExS, n=12) and exhaustive exercise with SAB (ExS+SAB, n=12). Five days before exhaustive exercise, SAB were medicated for 5 days in ExS+SAB group. Rats in ExS and ExS+SAB group were forced to swim for 150 minutes and then they were sacrificed, while rats in normal group were sacrificed at rest. After that, blood was collected and cardiac muscle tissue damage indices were analyzed. Results: Serum aspartate transaminase activity and lactate dehydrogenase activity were significantly lower in ExS+SAB group than in ExS group. Serum creatine phosphokinase activity of ExS+SAB was significantly lower than ExS group. However, the content of serum creatinine had no difference between ExS and ExS+SAB group. In the H&E stained left ventricle myocardium, ExS group showed signs of myocardial damage such as sporadic fragmentation of myocardial fibers, interstitial edema, cytoplasmic eosinophilia and neutrophils infiltration. However, ExS+SAB group alleviated the severity of the signs of myocardial damage. In the myocardial dihydroethidium staining, optical density was remarkably decreased in ExS+SAB group compared to ExS group. Furthermore, the up-regulation of Bax/Bcl-2 ratio was observed in ExS+SAB group compared with ExS group. Conclusions: The above results suggest that SAB may protect cardiac muscle damage via antioxidant activity and prevention of apoptosis.

Highly-sensitive Detection of Salvianolic Acid B using Alumina Microfibers-modified Electrode

Dong Sun,Xiaoyong Zheng,Xiafeng Xie,Xiaofeng Yang,Huajie Zhang 대한화학회 2013 Bulletin of the Korean Chemical Society Vol.34 No.11

Alumina microfibers with porous structures were prepared through hydrothermal reaction, and then used to modify the surface of carbon paste electrode (CPE). After modification with alumina microfibers, the electrochemical activity of CPE was found to be greatly improved. On the surface of alumina microfibersmodified CPE, the oxidation peak current of salvianolic acid B, a main bioactive compound in Danshen with anti-oxidative and anti-inflammatory effects, was remarkably increased compared with that on the bare CPE surface. The influences of pH value, amount of alumina microfibers and accumulation time were studied. Based on the strong signal amplification effects of alumina microfibers, a novel electrochemical method was developed for the detection of salvianolic acid B. The linear range was from 5 μg L−1 to 0.3 mg L−1, and the detection limit was 2 μg L−1 (2.78 nM) after 1-min accumulation. The new method was successfully used to detect salvianolic acid B in ShuangDan oral liquid samples, and the recovery was over the range from 97.4% to 102.9%.

달임 조건에 따른 Salvianolic Acid B의 구조변환 차이

이형재(Hyoung Jae Lee),조정용(Jeong-Yong Cho),이상현(Sang-Hyun Lee),전태일(Tae-Il Jeon),박근형(Keun-Hyung Park),문제학(Jae-Hak Moon) 한국식품과학회 2012 한국식품과학회지 Vol.44 No.6

식품 및 한약재는 수용액 중에서 가열 및 달임을 행한 후에 섭취하는 경우가 매우 많다. 그러나 가열 및 달임에 의한 소재 중에 함유된 성분변화 연구는 매우 미흡한 실정이다. 이에 본 연구에서는 순환기계질환 치료를 위해 많이 이용되고 있는 단삼(Salvia miltiorrhiza Bunge)이라는 한약재에 다량 함유된 salvianolic acid B(Sal B)를 대상으로 가열조건에 따른 성분변화 양상을 HPLC 분석을 통해 비교하였다. 그 결과 100℃의 동일 온도에서는 상압에서보다 가압조건 하에서 Sal B 변환 생성물들의 함량이 더 높아짐을 알 수 있었다. 그리고 가압조건 하에서 온도를 100℃로 동일하게 설정하고 달임시간에 따른 변화를 비교한 결과, 매우 한정적인 성분들을 제외하고, 대부분의 성분들이 가열시간의 경과와 함께 그 생성물들의 함량이 증가하는 경향을 보였다. 한편 가열 온도와 압력 조건을 달리하여 각각 4시간동안 가열을 행한 결과, 가열온도와 압력이 높을수록 생성률이 특징적으로 높아지는 생성물들이 있는 반면, 오히려 그 반대의 경우도 관찰되었다. 이 결과들로부터 과도한 가열 온도와 지나치게 높은 가압 조건에서는 상압조건과는 다른 가열생성물들의 조성이 형성됨을 알 수 있었다. 이에 따라 가열 방법 및 가열 조건에 따라 생리활성의 발현 양상에도 영향이 초래될 가능성이 시사되었다. 본 연구 결과는 한의학뿐만이 아니라 조리학적 측면에 있어서도 매우 중요한 성과라 판단되는 바 관련 연구의 기초자료로 활용되길 기대한다. The chemical conversion pattern of salvianolic acid B (Sal B) in aqueous solution under different boiling conditions was compared. When the duration of boiling was varied, without varying temperature or pressure, the content of chemically converted compounds (CCCs) was mostly increased over time. In addition, under different conditions of temperature and pressure with the same boiling time, the content of a few compounds increased with increasing temperature and pressure. These results confirmed that high temperatures and pressures in boiling alter the final composition of CCCs of Sal B. Therefore, it was suggested that the boiling conditions (time, temperature, and pressure) may be responsible for alteration of biological activities of the compounds. Our investigation of the chemical conversion of compounds contained in foods and medicinal herbs may provide important information in clarifying the biological activity of Sal B containing foods and medicinal herbs.