1.Determination of Silybin B in the Different Parts of Silybum marianum using HPLC-UV
Joyce P RODRIGUEZ ; Norman G QUILANTANG ; Ju Sung LEE ; Jeong Min LEE ; Hyun Young KIM ; Jae Suk SHIM ; Sanghyun LEE
Natural Product Sciences 2018;24(2):82-87
Silymarin is the standardized extract from Silybum marianum which consists mainly of flavonoids and polyphenols. It is highly regarded for its hepatoprotective ability. Silybin B is a flavonolignan and one of the active components of silymarin. The content of silybin B in various parts of S. marianum was analyzed by HPLC-UV. Results show that the extract of seeds contain the highest amount of silybin B (7.434 mg/g DW). The petioles of S. marianum showed a low content of silybin B. This study revealed that seeds of S. marianum contain high amount of silybin B and could be a good source of the compound.
Flavonoids
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Milk Thistle
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Polyphenols
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Silymarin
2.Identification of flavonoids 3-hydroxylase from Silybum marianum (L.) Gaertn and its application in enhanced production of taxifolin.
Song GAO ; Jingwen ZHOU ; Jian CHEN
Chinese Journal of Biotechnology 2020;36(12):2838-2849
(2S)-taxifolin is an important flavonoid that has anti-inflammatory and anti-oxidation effects. It is widely used in pharmaceutical and nutraceutical industries. Flavone 3-hydroxylase (F3H) can catalyze the synthesis of (2S)-taxifolin and other 3-hydroxylated flavonoids from (2S)-eriodictyol. Due to the low catalytic efficiency of F3H, the titer of many 3-hydroxyflavones, such as taxifolin, synthesized by microbial method is relatively low. In this study, a SmF3H was identified from the transcriptome of Silybum marianum (L.) Gaertn. The results of fermentation showed that SmF3H can catalyze the flavone 3-hydroxylation reaction, and its catalytic efficiency was significantly higher than that of commonly used SlF3H from Solanum lycopersicum. Six promoters with different transcription strength were selected to optimize the synthesis pathway from the flavonoid precursor (2S)-naringenin to (2S)-taxifolin. The results showed that the highest titer of (2S)-taxifolin (695.90 mg/L in shake flask) could be obtained when the P(GAL7) promoter was used to control the expression of SmF3H. The titer of (2S)-taxifolin was further improved to 3.54 g/L in a 5-L fermenter, which is the highest titer according to current available literatures.
Antioxidants
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Flavonoids
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Milk Thistle
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Quercetin/analogs & derivatives*
3.Luteolin 5-O-glucoside from Korean Milk Thistle, Cirsium maackii, Exhibits Anti-Inflammatory Activity via Activation of the Nrf2/HO-1 Pathway.
Hyun Ah JUNG ; Anupom ROY ; Qudeer Ahmed ABDUL ; Hyeung Rak KIM ; Hee Juhn PARK ; Jae Sue CHOI
Natural Product Sciences 2017;23(3):183-191
Luteolin 5-O-glucoside is the major flavonoid from Korean thistle, Cirsium maackii. We previously reported the anti-inflammatory activities of luteolin 5-O-glucoside in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. In this study, we determined the anti-inflammatory mechanisms of luteolin 5-O-glucoside through the inhibition of nitric oxide (NO) production in vitro and in vivo. Results revealed that luteolin 5-O-glucoside dose-dependently inhibited NO production and expression of iNOS and COX-2 in LPS-induced RAW 264.7 cells. Luteolin 5-O-glucoside also significantly inhibited the translocation of NF-κB, the activation of MAPKs, and ROS generation in LPS-induced RAW 264.7 cells. In addition, protein expressions of Nrf-2 and HO-1 were also upregulated by luteolin 5-O-glucoside treatment. Moreover, luteolin 5-O-glucoside inhibited λ-carrageenan-induced mouse paw edema by 65.34% and 48.31% at doses of 50 and 100 mg/kg body weight, respectively. These findings indicate potential anti-inflammatory effect of luteolin 5-O-glucoside particularly by downregulating NF-κB and upregulating HO-1/Nrf-2 pathway.
Animals
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Body Weight
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Cirsium*
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Edema
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In Vitro Techniques
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Luteolin*
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Mice
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Milk Thistle*
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Milk*
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Nitric Oxide
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RAW 264.7 Cells
4.Silibinin Enhances Ultraviolet B-Induced Apoptosis in MCF-7 Human Breast Cancer Cells.
Eun Mi NOH ; Mi Suk YI ; Hyun Jo YOUN ; Byoung Kil LEE ; Young Rae LEE ; Ji Hey HAN ; Hong Nu YU ; Jong Suk KIM ; Sung Hoo JUNG
Journal of Breast Cancer 2011;14(1):8-13
PURPOSE: Chemotherapies for breast cancer generally have strong cellular cytotoxicity and severe side effects. Thus, significant emphasis has been placed on combinations of naturally occurring chemopreventive agents. Silibinin is a major bioactive flavonolignan extracted from milk thistle with chemopreventive activity in various organs including the skin, prostate, and breast. However, the mechanism underlying the inhibitory action of silibinin in breast cancer has not been completely elucidated. Therefore, we investigated the effect of silibinin in MCF-7 human breast cancer cells and determined whether silibinin enhances ultraviolet (UV) B-induced apoptosis. METHODS: The effects of silibinin on MCF-7 cell viability were determined using the MTT assay. The effect of silibinin on PARP cleavage, as the hallmark of apoptotic cell death, and p53 protein expression in MCF-7 cells was analyzed using Western blot. The effect of silibinin on UVB-induced apoptosis in MCF-7 cells was analyzed by flow cytometry. RESULTS: A dose- and time-dependent reduction in viability was observed in MCF-7 cells treated with silibinin. Silibinin strongly induced apoptotic cell death in MCF-7 cells, and induction of apoptosis was associated with increased p53 expression. Moreover, silibinin enhanced UVB-induced apoptosis in MCF-7 cells. CONCLUSION: Silibinin induced a loss of cell viability and apoptotic cell death in MCF-7 cells. Furthermore, the combination of silibinin and UVB resulted in an additive effect on apoptosis in MCF-7 cells. These results suggest that silibinin might be an important supplemental agent for treating patients with breast cancer.
Apoptosis
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Blotting, Western
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Breast
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Breast Neoplasms
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Cell Death
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Cell Survival
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Humans
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MCF-7 Cells
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Milk Thistle
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Prostate
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Silymarin
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Skin
5.Inhibition of Wnt Signaling by Silymarin in Human Colorectal Cancer Cells.
Hyun Ji EO ; Gwang Hun PARK ; Jin Boo JEONG
Biomolecules & Therapeutics 2016;24(4):380-386
Silymarin from milk thistle (Silybum marianum) has been reported to show an anti-cancer activity. In previous study, we reported that silymarin induces cyclin D1 proteasomal degradation through NF-κB-mediated threonine-286 phosphorylation. However, mechanism for the inhibition of Wnt signaling by silymarin still remains unanswered. Thus, we investigated whether silymarin affects Wnt signaling in human colorectal cancer cells to elucidate the additional anti-cancer mechanism of silymarin. Transient transfection with a TOP and FOP FLASH luciferase construct indicated that silymarin suppressed the transcriptional activity of β-catenin/TCF. Silymarin treatment resulted in a decrease of intracellular β-catenin protein but not mRNA. The inhibition of proteasome by MG132 and GSK3β inhibition by SB216763 blocked silymarin-mediated downregulation of β-catenin. In addition, silymarin increased phosphorylation of β-catenin and a point mutation of S33Y attenuated silymarin-mediated β-catenin downregulation. In addition, silymarin decreased TCF4 and increased Axin expression in both protein and mRNA level. From these results, we suggest that silymarin-mediated downregulation of β-catenin and TCF4 may result in the inhibition of Wnt signaling in human colorectal cancer cells.
Colorectal Neoplasms*
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Cyclin D1
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Down-Regulation
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Humans*
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Luciferases
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Milk Thistle
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Phosphorylation
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Point Mutation
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Proteasome Endopeptidase Complex
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RNA, Messenger
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Silymarin*
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Transfection
6.Silymarin Inhibits Cytokine-Stimulated Pancreatic Beta Cells by Blocking the ERK1/2 Pathway.
Eun Jeong KIM ; Jeeho KIM ; Min Young LEE ; Muddenahalli Srinivasa SUDHANVA ; Sundaravinayagam DEVAKUMAR ; Young Jin JEON
Biomolecules & Therapeutics 2014;22(4):282-287
We show that silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), inhibits cytokine mixture (CM: TNF-alpha, IFN-gamma, and IL-1beta)-induced production of nitric oxide (NO) in the pancreatic beta cell line MIN6N8a. Immunostaining and Western blot analysis showed that silymarin inhibits iNOS gene expression. RT-PCR showed that silymarin inhibits iNOS gene expression in a dose-dependent manner. We also showed that silymarin inhibits extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) phosphorylation. A MEK1 inhibitor abrogated CM-induced nitrite production, similar to silymarin. Treatment of MIN6N8a cells with silymarin also inhibited CM-stimulated activation of NF-kappaB, which is important for iNOS transcription. Collectively, we demonstrate that silymarin inhibits NO production in pancreatic beta cells, and silymarin may represent a useful anti-diabetic agent.
Blotting, Western
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Gene Expression
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Insulin-Secreting Cells*
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Milk Thistle
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NF-kappa B
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Nitric Oxide
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Phosphorylation
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Silymarin*
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Tumor Necrosis Factor-alpha
7.Protective Effects of Silibinin and Its Possible Mechanism of Action in Mice Exposed to Chronic Unpredictable Mild Stress.
Wen Jing YAN ; Ying Chun TAN ; Ji Cheng XU ; Xian Ping TANG ; Chong ZHANG ; Peng Bo ZHANG ; Ze Qiang REN
Biomolecules & Therapeutics 2015;23(3):245-250
Silibinin, a natural flavonoid antioxidant isolated from extracts of the milk thistle herb, has recently been identified as having anti-hepatotoxic and anticancer properties. In this paper, we investigated the effects of silibinin on behavior and neuroplasticity in mice subjected to chronic unpredictable mild stress (CUMS). After 5 consecutive weeks of CUMS, the mice were treated with silibinin (100 mg/kg, 200 mg/kg and 400 mg/kg by oral gavage) for 3 consecutive weeks. The results showed that silibinin administration significantly alleviated the CUMS-induced depressive-like behavior, including the total number of squares crossed and the frequency of rearing in the open field test, the immobility time in the tail suspension test and the forced swimming test. Furthermore, silibinin treatment increased the levels of brain-derived neurotrophic factor (BDNF), serotonin (5-HT) and norepinephrine (NE) in the prefrontal cortex and hippocampus. Our study provides new insight into the protective effects of silibinin on the depressive status of CUMS mice, specifically by improving neuroplasticity and neurotransmission.
Animals
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Brain-Derived Neurotrophic Factor
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Depression
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Hindlimb Suspension
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Hippocampus
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Mice*
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Milk Thistle
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Neuronal Plasticity
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Norepinephrine
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Physical Exertion
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Prefrontal Cortex
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Serotonin
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Synaptic Transmission
8.Beneficial Effects of Silibinin Against Kainic Acid-induced Neurotoxicity in the Hippocampus in vivo.
Sehwan KIM ; Un Ju JUNG ; Yong Seok OH ; Min Tae JEON ; Hyung Jun KIM ; Won Ho SHIN ; Jungwan HONG ; Sang Ryong KIM
Experimental Neurobiology 2017;26(5):266-277
Silibinin, an active constituent of silymarin extracted from milk thistle, has been previously reported to confer protection to the adult brain against neurodegeneration. However, its effects against epileptic seizures have not been examined yet. In order to investigate the effects of silibinin against epileptic seizures, we used a relevant mouse model in which seizures are manifested as status epilepticus, induced by kainic acid (KA) treatment. Silibinin was injected intraperitoneally, starting 1 day before an intrahippocampal KA injection and continued daily until analysis of each experiment. Our results indicated that silibinin-treatment could reduce seizure susceptibility and frequency of spontaneous recurrent seizures (SRS) induced by KA administration, and attenuate granule cell dispersion (GCD), a morphological alteration characteristic of the dentate gyrus (DG) in temporal lobe epilepsy (TLE). Moreover, its treatment significantly reduced the aberrant levels of apoptotic, autophagic and pro-inflammatory molecules induced by KA administration, resulting in neuroprotection in the hippocampus. Thus, these results suggest that silibinin may be a beneficial natural compound for preventing epileptic events.
Adult
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Animals
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Brain
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Dentate Gyrus
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Epilepsy
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Epilepsy, Temporal Lobe
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Hippocampus*
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Humans
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Kainic Acid
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Mice
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Milk Thistle
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Neuroprotection
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Seizures
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Silymarin
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Status Epilepticus
9.Determination of concentration and encapsulation efficiency of silybin liposomes.
Jiang-nan YU ; Xi-ming XU ; Yuan ZHU ; Hao GAO
China Journal of Chinese Materia Medica 2003;28(11):1027-1030
OBJECTIVETo determine the encapsulation efficiency of silybin liposomes.
METHODAfter Silybin liposomes solution was separated through sphadex G-100 column, eluent collected was determined by the first derivative UV spectrometry.
RESULTThe calibration curve of Silybin was linear in the range of 0.5-30.0 mg x L(-1) (r = 0.9998). The average recovery was 101.7%. The relative standard deviation was 1.8%. The results showed that the encapsulation efficiency ranged from 65.1%-83.0% when the drug/phospholipid ratio varied from 0.02-0.06. The concentration of SLB in liposomes prepared by optimum technique was 760.4 mg x L(-1) (n = 4).
CONCLUSIONThe method is accurate, simple and suitable for the quality control of Silybin liposomes.
Drug Delivery Systems ; Liposomes ; chemistry ; Milk Thistle ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Silymarin ; administration & dosage ; analysis ; isolation & purification ; Spectrophotometry, Ultraviolet ; Technology, Pharmaceutical ; methods
10.Establishment of culture system of Silybum marianum hairy roots and determination of silybin.
Shu-Li ZHANG ; Tian-Zhu ZHANG ; Shi-Hai YANG
China Journal of Chinese Materia Medica 2014;39(11):2005-2010
This research uses six Agrobacterium rhizogenes R1601, R15384, R1000, A4, R1025 and R1 to infect silymarin explants to induce hairy roots and silibin. All of the six A. rhizogenes can induce Silybum marianum to generate hairy roots and the A. rhizogene A4 shows comparatively high infection on the plant. This research determines the condition to induce silymarin hairy roots by the factors of infection time, pre-culturing, co-culturing and pH value. The fact that MS liquid medium fits the proliferation of silymarin hairy roots is determined. Through PCR molecular identification, it can be seen that the DNA plasmids in the A. rhizogenes are successfully integrated into the genome of transformed roots. Using liquid chromatography, it is determined that the silibin content in silymarin hairy roots is 2.5 times that in the plant In this research, the silymarin hairy roots culturing system is established, which lays a foundation for the study of culturing silymarin hairy roots and producing silibin.
Agrobacterium
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genetics
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physiology
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Cell Culture Techniques
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methods
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Milk Thistle
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chemistry
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genetics
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growth & development
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microbiology
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Plant Roots
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chemistry
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genetics
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growth & development
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microbiology
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Silymarin
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analysis
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Transformation, Genetic