Natural flavonolignan from milk thistle (Silybum marianum) seeds

水飞蓟宾的生长抑制具有时间和剂量依赖性（0-200 mM；持续 72 小时）[1]。水飞蓟宾（68 μM；持续 72 小时）会导致细胞凋亡，并将 G1 期细胞的百分比提高至约 22% [1]。水飞蓟宾 (68 μM) 会诱导 AKT 活性抑制，持续时间为 72 小时[1]。

---

是从水飞蓟中提取的黄酮木脂素，具有保肝、抗氧化和抗炎活性。几项研究表明，水飞蓟宾对预防和治疗不同类型的癌症非常有效，其抗肿瘤机制包括阻止细胞周期和/或细胞凋亡。采用MTT法研究细胞活力，分别采用硫代巴比妥酸活性物质（TBARS）法、NO法和MnSOD法研究脂质过氧化、细胞外NO生成和清除酶活性。流式细胞仪进行细胞周期和凋亡分析。实时PCR检测miRNA谱。在本研究中，我们证明了水飞蓟宾诱导的生长抑制在细胞周期的G1期阻断Hepg2细胞并激活细胞程序性死亡过程。此外，水飞蓟宾的抗增殖作用与参与miRNA分泌调节的神经酰胺数量的强烈增加相平行。特别是水飞蓟宾处理后，miR223-3p和miR16-5p上调，miR-92-3p下调（p < 0.05）。综上所述，我们的研究结果表明，水飞蓟宾诱导的HepG2细胞凋亡与神经酰胺合成和miRNAs分泌的增加同步发生。

在过去的四个星期中，水飞蓟宾（50-100 mg/kg/天；胃内给药）已被证明可以显着减少肝脏和血清脂质积累[2]。

---

水飞蓟宾对肥胖和代谢综合征有良好的作用，但水飞蓟宾的全身调节作用尚未完全揭示。本研究旨在探讨水飞蓟宾对非酒精性脂肪性肝病（NAFLD）代谢的调节作用。C57BL/6 J小鼠连续8周饲喂高脂高胆固醇饲粮，后4周灌胃水飞蓟素（50或100 mg/kg/d）和牛磺酸脱氧胆酸钠（50 mg/kg/d）。采用血液生化指标、肝脏脂质测定及肝脏油红O染色评价水飞蓟宾和TUDCA的模型及降脂效果。此外，血清和肝脏样品通过基于气相色谱-质谱（GC/MS）的代谢组学平台进行检测。多变量/单变量数据分析和途径分析用于研究差异代谢物和代谢途径。结果表明，小鼠NAFLD模型成功建立，水飞蓟宾和TUDCA均能显著降低血清和肝脏脂质积累。血清和肝脏代谢组学分析表明，高脂/高胆固醇饮食导致脂质代谢、多元醇代谢、氨基酸代谢、尿素循环和TCA循环等代谢产物代谢异常。水飞蓟宾和TUDCA处理均能逆转HFD喂养引起的代谢紊乱。综上所述，高脂肪/高胆固醇饮食导致小鼠血清和肝脏代谢异常，水飞蓟宾治疗改善了肝脏脂质积累，调节了整体代谢途径，这可能解释了其多靶点机制[2]。

细胞活力测定[1]
细胞类型： HepG2 细胞生长
测试浓度： 0-200 mM
孵育时间： 72 小时
实验结果： 以时间和剂量依赖性方式抑制生长，IC50 为 68 μM。

---

细胞凋亡分析[1]
细胞类型： HepG2 细胞生长
测试浓度： 68 μM
孵育时间：72小时
实验结果：与未处理的细胞相比，诱导更多细胞凋亡（60%）。

---

细胞周期分析[1]
细胞类型： HepG2 细胞生长
测试浓度： 68 μM
孵育持续时间： 72 小时
实验结果： G1 期细胞增加约 22%，S 期细胞减少 47%。

---

蛋白质印迹分析[1]
细胞类型： HepG2 细胞生长
测试浓度： 68 μM
孵育持续时间： 72 小时
实验结果： 诱导 AKT 活性抑制。

Animal/Disease Models: Male C57BL/6J mice (6-8 weeks old) with nonalcoholic fatty liver disease (NAFLD)[2]
Doses: 50, 100 mg/kg
Route of Administration: Given intragastrically (po); daily; for the last 4 weeks
Experimental Results: Dramatically lowered both serum and hepatic lipid accumulation.

---

Male C57BL/6J mice (6–8 weeks old) were acclimatized under 12 h/12 h dark-light cycles at a constant temperature (22 ± 2℃) and had free access to water and food. All mice were fed a normal diet for one week to acclimate to the environment. After that, they were divided into 5 groups (n = 6): vehicle group, HFD (high-fat/high-cholesterol diet) group, LS (low-dose Silybin) group, HS (high-dose Silybin) group and TUDCA group. The vehicle group was continuously fed a standard normal diet, and the other groups were fed a high-fat/high-cholesterol diet (10 % lard, 10 % yolk, 1 % cholesterol, 0.2 % cholate and 78.8 % standard diet; 60 % of kcal as fat was the prescription) for 8 weeks. The standard normal diet and high-fat/high-cholesterol diet were both obtained from Nanjing Qinglongshan Experimental Animal Center. Silybin (50 or 100 mg/kg/day) and TUDCA (50 mg/kg/day) were ground in 0.5 % carboxymethylcellulose sodium (CMC-Na) and were given intragastrically for the last 4 weeks. At the end of the experiment, blood was collected from the orbital sinus after fasting overnight, and the levels of serum total cholesterol (TC), triglyceride (TG) and nonesterified fatty acid (NEFA) were assayed with commercial kits purchased from xxx. Livers were frozen by dry ice in OCT® compound during tissue collection and then sectioned into 8-μm-thick slices by a cryostat and stained with Oil red O as previously described. Both serum and liver samples were stored at -80 °C until analysis.[2]

Metabolism / Metabolites
Silybin has known human metabolites that include O-demethylated-silybin.

[1]. Silybin-Induced Apoptosis Occurs in Parallel to the Increase of Ceramides Synthesis and miRNAs Secretion in Human Hepatocarcinoma Cells. Int J Mol Sci. 2019 May 3;20(9):2190.

[2]. Silybin ameliorates hepatic lipid accumulation and modulates global metabolism in an NAFLD mouse model. Biomed Pharmacother. 2020 Mar;123:109721.

Silibinin is a flavonolignan isolated from milk thistle, Silybum marianum, that has been shown to exhibit antioxidant and antineoplastic activities. It has a role as an antioxidant, an antineoplastic agent, a hepatoprotective agent and a plant metabolite. It is a flavonolignan, a polyphenol, an aromatic ether, a benzodioxine and a secondary alpha-hydroxy ketone.
Silibinin is the major active constituent of silymarin, a standardized extract of the milk thistle seeds, containing a mixture of flavonolignans consisting of silibinin, isosilibinin, silicristin, silidianin and others. Silibinin is presented as a mixture of two diastereomers, silybin A and silybin B, which are found in an approximately equimolar ratio. Both in vitro and animal research suggest that silibinin has hepatoprotective (antihepatotoxic) properties that protect liver cells against toxins. Silibinin has also demonstrated in vitro anti-cancer effects against human prostate adenocarcinoma cells, estrogen-dependent and -independent human breast carcinoma cells, human ectocervical carcinoma cells, human colon cancer cells, and both small and nonsmall human lung carcinoma cells.
Silibinin has been reported in Aspergillus iizukae, Silybum eburneum, and other organisms with data available.
Silymarin is a mixture of flavonolignans isolated from the milk thistle plant Silybum marianum. Silymarin may act as an antioxidant, protecting hepatic cells from chemotherapy-related free radical damage. This agent may also promote the growth of new hepatic cells. (NCI04)
The major active component of silymarin flavonoids extracted from seeds of the MILK THISTLE, Silybum marianum; it is used in the treatment of HEPATITIS; LIVER CIRRHOSIS; and CHEMICAL AND DRUG INDUCED LIVER INJURY, and has antineoplastic activity; silybins A and B are diastereomers.

---

Drug Indication
Currently being tested as a treatment of severe intoxications with hepatotoxic substances, such as death cap (Amanita phalloides) poisoning.

---

In conclusion, these data suggest that silybin through the synthesis of ceramides may to activate the processes of programmed death by inducing the secretion of specific miRNAs that likely target (PTEN)/AKT pathway. These findings offer new therapies to treat cancer that is poorly sensitive to pharmacological treatments, such as HCC at an advanced stage. The aim of this study was to investigate in depth the molecular mechanisms of silybin antitumor efficacy in a noninvasive and faster way with respect to all the in vivo procedures.[1]

---

In conclusion, NAFLD, at least in our model, showed significantly increased lipid accumulation in the serum and liver accompanied by a global abnormal metabolism involving lipid metabolism, polyol metabolism, amino acid metabolism, the urea cycle and the TCA cycle. TUDCA significantly modulated these metabolic pathways. Silybin had a marked modulation effect on these metabolic pathways, and the modulated pathways were consistent with its multiple target mechanism. Although most metabolites involved in energy metabolism, such as carbohydrates, amino acids and fatty acids, were detected by GC–MS, there was a lack of more comprehensive coverage of endogenous metabolites. Furthermore, a mechanism study of these pathways modulated by silybin based on molecular biology needs to be further conducted.[2]

C25H22O10

482.44

482.121

802918-57-6

Silybin A;22888-70-6;Isosilybin;72581-71-6;Silybin B;142797-34-0

31553

White to off-white solid powder

152-153°C

2.362

155.14

5

10

4

35

750

4

COC1=C(C=CC(=C1)[C@@H]2[C@H](OC3=C(O2)C=C(C=C3)[C@@H]4[C@H](C(=O)C5=C(C=C(C=C5O4)O)O)O)CO)O

SEBFKMXJBCUCAI-HKTJVKLFSA-N

InChI=1S/C25H22O10/c1-32-17-6-11(2-4-14(17)28)24-20(10-26)33-16-5-3-12(7-18(16)34-24)25-23(31)22(30)21-15(29)8-13(27)9-19(21)35-25/h2-9,20,23-29,31H,10H2,1H3/t20-,23+,24-,25-/m1/s1

(2R,3R)-3,5,7-trihydroxy-2-[(2R,3R)-3-(4-hydroxy-3-methoxyphenyl)-2-(hydroxymethyl)-2,3-dihydro-1,4-benzodioxin-6-yl]-2,3-dihydrochromen-4-one

36804-17-8; 678-483-8; 802918-57-6; Legalon; SILYMARIN; Silybin (Standard); CHEMBL9509;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 25 mg/mL (51.82 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.18 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL 澄清 DMSO 储备液加入900 μL 玉米油中，混合均匀。

---

请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。