Hepatoprotective effect of obeticholic acid on acetaminophen induced hepatotoxicity in mice

Acetaminophen (APAP) is commonly used as analgesic and antipyretic drug for relieving mild and moderate pain, but at high doses produces hepatic necrosis. Though, Obeticholic acid (OCA) has been tested in range of diseases, its therapeutic potential against APAP-induced hepatic injury remains to be elucidated. Thus, in this study, we investigated the preventive effect of OCA along with N-acetylcysteine (NAC) and Silymarin (SIL) against acetaminophen-induced hepatotoxicity in mice. SIL (100 mg/kg, po) and OCA (30 mg/kg, po) were administered continuously for six days prior to APAP administration. After sixth dose, animas were fasted for 12 h and treated with 300 mg/kg APAP and then received SIL (100 mg/kg, po), NAC (500 mg/kg, ip) and OCA (30 mg/kg, po) at 1 h after APAP. Mice were sacrificed 6 h after APAP injection. Analysis of serum Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Alkaline phosphatase (ALP), liver glutathione (GSH) and histopathology were employed for assessment of hepatotoxicity. APAP group showed a significant increase in ALT, AST, ALP and centriolobular hepatic necrosis with a significant decrease in glutathione in comparison to control group. All these parameters were significantly improved in all the three treated groups when compared to APAP group. In conclusion, Obeticholic acid (OCA), Silymarin (SIL) and N-acetylcysteine (NAC) are suggested to protect against APAP-induced hepatotoxicity in mice by ameliorating liver enzymes, antioxidant effect and decreasing liver necrosis.

Network pharmacology study of flavone extracts of Silybum marianum in the treatment of nonalcoholic fatty liver disease / 药学学报

To investigate the therapeutic effect and molecular mechanism of the main flavonoid components of Silybum marianum (S. marianum) on nonalcoholic fatty liver disease (NAFLD), we identified nine flavonoids in S. marianum through TCMSP, PubChem database and corresponding literatures. The potential therapeutic targets of NAFLD were predicted by SwissTargetPrediction, GeneCards and Venny 2.1.0 platform, while the protein-protein interaction (PPI) network of potential targets was analyzed using String platform and Cytoscape software. Then GO and KEGG pathway enrichment analysis were performed using David 6.8 database, followed by molecular docking verification using AutoDock software. In vitro, components with higher degree value in the "components-targets-pathway" network were chosen for further analysis. L02 cells were used to establish lipid accumulation model and treated with different components. Furthermore, the effects of four pure active compounds from S. marianum on lipid accumulation in hepatocytes were analyzed by oil red O staining. The results showed that the main nine flavonoids extracted from S. marianum contained 24 potential NAFLD targets. Several critical pathways closely related to NAFLD process were identified by GO and KEGG enrichment analysis, including phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) pathway, type 2 diabetes pathway, tumor necrosis factor (TNF) pathway and insulin resistance pathway. The results of molecular docking further indicated that the core components displayed strong binding abilities with key targets respectively, and silandrin showed better binding activity as compared to other components. The results obtained from L02 cells showed that the lipid accumulation was reduced by treatment with isosilybin A, isosilybin B, silydianin and silychristin, while the activity of isosilybin B was better than that of isosilybin A. Taken together, we concluded that the main flavone components of S. marianum could improve lipid accumulation through multiple signaling pathway in hepatocytes, and this could be a potential new strategy for the treatment of NAFLD.

Efficacy of an herbal compound in decreasing steatosis and transaminase activities in non-alcoholic fatty liver disease: A randomized clinical trial

Abstract Hepatoprotective effects of many herbal agents have been reported in animal studies and clinical trials. In this study, five hepatoprotective plants with potent antioxidant, anti-inflammatory, and hypolipidemic effects were chosen to prepare a polyherbal compound for managing NAFLD. Sixty patients with NAFLD were randomly divided into treatment and control groups (2:1 ratio). Both group were advised to take healthy diet and exercise. The treatment group also received herbal capsules containing 400 mg of the mixture of Anethum graveolens, Citrus aurantium, Cynara scolymus, Portulaca oleracea, and Silybum marianum (2 capsules, thrice daily, for two months). The liver ultrasound and biochemical markers including the serum lipids, liver enzymes, and glucose were evaluated before starting the study and at the end of the treatment. Thirty patients in the treatment group and sixteen patients in the control group completed the study. The herbal compound significantly decreased the serum level of alanine transaminase (ALT), aspartate transaminase (AST), and total cholesterol. Treatment with the herbal compound significantly improved the grade of the fatty liver, but no significant change was found in the control group. In conclusion, the formulated herbal compound appeared to be effective in biochemical improvement and decreasing the grade of the fatty liver in the patients with NAFLD.

The benefit of Silybum marianum in ethanol-induced reprotoxicity of male Wistar rat

Abstract This study investigates the toxic effects of ethanol (Eth) on the reproductive system of male rats and the possible protective role of Silybum marianum seeds-infused solution (SMI) over six consecutive weeks of administration. Animals were divided into the following groups: control, SMI positive control (200 mg/kg/day), Eth1 (1 g/kg/day), Eth2 (2 g/kg/day), Eth1+SMI, and Eth2+SMI. Plasma testosterone concentration, epididymal spermatozoa biology, and testicular and epididymal MDA, GSH and GPx levels were evaluated. The results indicated a significant decrease in testis and epididymis weight, testosterone level, sperm concentration, sperm vitality and sperm motility (total motility, progressive motility, curvilinear velocity, straight-line velocity, velocity average path, beat cross frequency, and lateral head displacement) in both Eth1 and Eth2 compared to the control groups and the combined-treatment groups (Eth1+SMI and Eth2+SMI). Furthermore, results showed a significant elevation in MDA concentration with a significant decrease of testicular and epididymal GSH concentration and GPx activity in theEth1 and Eth2 groups compared to the combined-treatment groups. The administration of SMI succeeded in improving the parameters cited above in the combined-treatment groups compared to the Eth1 and Eth2 groups, and bring them to the levels seen in the control groups. To conclude, SMI has clearly protected reproductive indices against ethanol-induced reprotoxicity in male rats

Potential effect of Silybum marianum L. and Cistus ladaniferus L. extracts on urine volume, creatinine clearance and renal function

Objective:To investigate the diuretic and renal effects of Silybum marianum L. and Cistus ladaniferus L. in normal rats.Methods:Four groups of rats were used in each experiment. The first group received water, the second group received Cistus ladaniferus L. extract (100 mg/kg b.wt), the third group received Silybum marianum L. extract (100 mg/kg b.wt), and the fourth group received furosemide (10 mg/kg b.wt). Variables including urine volume, plasma and urine sodium, potassium and creatinine, and creatinine clearance were measured. Two experiments were conducted. A single dose of each intervention was used and the variables were measured during 24 h, and the interventions were given daily for a total of 8 d and the variables were measured during various intervals.Results:The single dose of each plant extract increased urine volume at all-time intervals and increased urine sodium and potassium excretion without affecting plasma sodium and potassium (P<0.05). On the day 8 after daily administration, the plant extracts induced a significant diuresis and natriuresis without affecting serum electrolytes (P<0.05), while furosemide caused hypokalemia. Both plant extracts significantly increased creatinine clearance (P<0.05).Conclusions:Silybum marianum L. and Cistus ladaniferus L. increase creatinine clearance and have a significant diuretic effect without affecting serum electrolytes. Silybum marianum L. is more potent than furosemide or Cistus ladaniferus L.

Determination of Silybin B in the Different Parts of Silybum marianum using HPLC-UV

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.

Potential effect of Silybum marianum L. and Cistus ladaniferus L. extracts on urine volume, creatinine clearance and renal function

Objective: To investigate the diuretic and renal effects of Silybum marianum L. and Cistus ladaniferus L. in normal rats. Methods: Four groups of rats were used in each experiment. The first group received water, the second group received Cistus ladaniferus L. extract (100 mg/kg b.wt), the third group received Silybum marianum L. extract (100 mg/kg b.wt), and the fourth group received furosemide (10 mg/kg b.wt). Variables including urine volume, plasma and urine sodium, potassium and creatinine, and creatinine clearance were measured. Two experiments were conducted. A single dose of each intervention was used and the variables were measured during 24 h, and the interventions were given daily for a total of 8 d and the variables were measured during various intervals. Results: The single dose of each plant extract increased urine volume at all-time intervals and increased urine sodium and potassium excretion without affecting plasma sodium and potassium (P<0.05). On the day 8 after daily administration, the plant extracts induced a significant diuresis and natriuresis without affecting serum electrolytes (P<0.05), while furosemide caused hypokalemia. Both plant extracts significantly increased creatinine clearance (P<0.05). Conclusions: Silybum marianum L. and Cistus ladaniferus L. increase creatinine clearance and have a significant diuretic effect without affecting serum electrolytes. Silybum marianum L. is more potent than furosemide or Cistus ladaniferus L. http://www.apjtm.org/article.asp?issn=1995-7645;year=2018;volume=11;issue=6;spage=393;epage=398;aulast=El;type=2.

Anti-inflammatory effect of silymarin on ovarian immunohistochemical localization of TNF-alpha associated with systemic inflammation in polycystic ovarian syndrome / Efecto antiinflamatorio de la silimarina en la localización inmunohistoquímica ovárica del TNF-alfa asociado con la inflamación sistémica en el síndrome de ovario poliquístico

Tumor necrosis factor alpha (TNF-a) and interleukin (IL)-6, are prominent mediators of inflammation and have been confirmed to be elevated in at least a subgroup of women with polycystic ovary syndrome (PCOS). In this study, the effects of Silymarin (SLM) on the expression TNF-a, IL-6, CRP and symptoms of PCOS were studied. In this research, PCOS was induced by injection of Estradiol Valerate. PCOS rats were divided into control and experimental groups received intraperitoneal injection SLM extract daily. After syndrome induction, ovaries were collected for histological and immunohistochemical evaluations. Serum IL-6 was detected by the ELISA kit. The results indicated the significant reduction in inflammatory markers and significant changes follicular layers thickness in the treatment group as compared with control. It can be concluded that having anti-inflammatory substances, Silymarin is effective in symptoms of this syndrome and metabolic syndrome.

El factor de necrosis tumoral alfa (TNF-a) y la interleucina (IL) -6 son mediadores prominentes de la inflamación y se ha confirmado que están elevados en al menos un subgrupo de mujeres con síndrome de ovario poliquístico (SOP). En este estudio se estudiaron los efectos de Silymarin (SLM) en la expresión TNF-a, IL-6, PCR y síntomas de SOP. El SOP fue inducido por inyección de valerato de estradiol. Las ratas SOP se dividieron en grupos control y los grupos experimentales recibieron diariamente un extracto de SLM por inyección intraperitoneal. Después de la inducción del síndrome, los ovarios se analizaron mediante histología e inmunohistoquímica. Se detectó IL-6 en suero mediante el kit ELISA. Los resultados indicaron una reducción significativa en los marcadores inflamatorios y cambios significativos en el espesor de las capas foliculares en el grupo de tratamiento en comparación con el control. Se puede concluir que con sustancias anti-inflamatorias, Silymarin es eficaz en los síntomas de este síndrome y el síndrome metabólico.

Antioxidant and Anti-aging Activities of Silybum Marianum Protein Hydrolysate in Mice Treated with D-galactose / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In the present study, we investigated the antioxidant and anti-aging effects of Silybum marianum protein hydrolysate (SMPH) in D-galactose-treated mice.</p><p><b>METHODS</b>D-galactose (500 mg/kg body weight) was intraperitoneally injected daily for 7 weeks to accelerate aging, and SMPH (400, 800, 1,200 mg/kg body weight, respectively) was simultaneously administered orally. The antioxidant and anti-aging effects of SMPH in the liver and brain were measured by biochemical assays. Transmission electron microscopy (TEM) was performed to study the ultrastructure of liver mitochondri.</p><p><b>RESULTS</b>SMPH decreased triglyceride and cholesterol levels in the D-galactose-treated mice. It significantly elevated the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC), which were suppressed by D-galactose. Monoamine oxidase (MAO) and malondialdehyde (MDA) levels as well as the concentrations of caspase-3 and 8-OHdG in the liver and brain were significantly reduced by SMPH. Moreover, it increased Bcl-2 levels in the liver and brain. Furthermore, SMPH significantly attenuated D-galactose-induced liver mitochondrial dysfunction by improving the activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase as well as mitochondrial membrane potential (ΔΨm) and fluidity. TEM showed that the degree of liver mitochondrial damage was significantly decreased by SMPH.</p><p><b>CONCLUSION</b>The results indicated that SMPH protects against D-galactose-induced accelerated aging in mice through its antioxidant and anti-aging activities.</p>

Bioactive lignans from Silybum marianum / 中国中药杂志

The seeds of Silybum marianum were extracted by hot water, and the extract was isolated by D101 macroporous resin, MCI resin, MPLC, HPLC, et al. As a result, 7 compounds including tricin 4'-O-[threo-β-guaiacyl-(7″-O-methyl)-glyceryl] ether(1), tricin 4'-O-[erythro-β-guaiacyl-(7″-O-methyl)-glyceryl] ether(2), 5'-methoxyhydnocarpin-D(3),palstatin(4),(8R,7'S,8'R)-5,5'-dimethoxy-7-oxolariciresinol 9'-O-D-xylopyranoside(5), 9-O-D-glucopyranoside(6), and(-)-haplomyrtoside(7) were isolated and identified for the first time. Compounds 1, 3, 4, and 5 exhibited activity against influenza A(H5N1)with IC₅₀ value of 0.65, 0.21, 0.32, and 0.56 μmol•L⁻¹, respectively. Compounds 1, 2, 6, and 7 exhibited cytotoxity against HepG-2 with IC₅₀ value of 0.35, 0.25, 0.53, 0.66 μmol•L⁻¹, respectively.

Establishment of Silybum marianum cell suspension culture system and effects of different factors on silybin content / 中草药

To establish the suspension culture system of Silybum marianum cell and study the effects of different factors on silybin content. Orthogonal test was adopted to determine S. marianum cell suspension culture system and HPLC to silybin content.. The best growth cycle of cell suspension culture was defined when cultured in liquid MS medium with 6-BA 1.5 mg/L + NAA 1.0 mg/L + ZT 1.5 mg/L, light 12 h/d, pH value = 7, revolution = 110 r/min. The effects of salicylic acid, chitosan, and sodium nitroprusside on cell suspension culture were investigated. When the concentration of three impact factors were 0.05, 3, and 1 μ g/L, respectively, the growth amount of S. marianum cell reached the maximum, i. e. At the same time, the silybin content also reached the maximum. The established cell suspension culture system is suitable to use for the rapid propagation of S. marianum. Chitosan and sodium nitroprusside at proper concentration are benifit to the growth of suspensious cells and accumulation of silybin.

Study of quality testing method for seeds of Silybum marianum / 中草药

Objective: To study the quality testing method for the seeds of Silybum marianum, so as to provide the basis for the development of testing procedures and quality grading standard for the seeds of S. marianum. Methods: Refering to "The International Seed Testing Rules" and "China Crop Seed Testing Rules" to carry out the quality testing for the seeds of S. marianum. Results: The seed cleanliness was analyzed by winnowing method; The authenticity was identified by morphological appearance compared with 1000 grain weight by 1000 grain weight determination method; The germination conditions for seed germination were washed with running tap water before 2 h, the double filter paper was used as germination bed, and the seeds were incubated at 20 ℃, 8000 lx light, counting time for 4-14 d; The viability was determined by electrical conductivity method. Conclusion: The method is simple and reliable to the quality testing for the seeds of S. marianum.

Isolation of Aliphatic Carbonyl Hydrocarbons and in-vitro Antioxidant Activity of Silybum Marianum Seeds.

Silybum marianum (milk thistle) seeds and other parts of this medicinal plant have been used traditionally for centuries in the treatment of liver and several other diseases. Purified extracts made from the fruits of Silybum marianum (SM) chiefly contains silymarin and are available commercially. Silymarin is used as a dietary supplement and is one of the widely used alternative medicines. The present study was conducted to investigate the in- vitro antioxidant potential of SM seed extracts by DPPH method and to isolate the phytoconstituents present in ethyl acetate extract. Phytochemical investigation of the seeds of SM yielded eight aliphatic carbonyl hydrocarbons. The chemical structure of all the phytoconstituents were elucidated on the basis of spectral data analysis. Antioxidant activity of methanolic seed extract was found to be better than the ethyl acetate as well as standard ascorbic acid which could be due to high flavonoid content. Thus result of this study support use of silymarin as a dietary natural antioxidant supplement in drug and food industry to prevent free radical related diseases.

Effects of exogenous nitric oxide on physiology of seed germination and seedling growth of Silybum marianum under NaCl stress / 中草药

Objective: To investigate the effects of exogenous nitric oxide (NO) on the physiology of the seed germination and seedling growth of Silybum marianum under NaCl stress. Methods: The seeds of S. marianum were treated by sodium nitro prusside (SNP) at the concentration of 0.05-0.60 mmol/L under 0.7% NaCl stress. Some physiological indexes were measured, such as germination energy, germination rate, germination index, and vigor index of the seeds, and contents of malondialdehyde (MDA), photosynthetic pigment, osmosis substances, and the activities of the protective enzymes in leaves. Results: The seed germination and seedling growth of S. marianum were obviously inhibited under NaCl stress. Soaking seeds with 0.05-0.60 mmol/L SNP could alleviate the damage of NaCl stress. Under this treatment, the contents of photosynthetic pigment (including chlorophyll a, chlorophyll b, total chlorophyll, and carotinoid) and osmosis substances (including soluble sugar, soluble protein, and proline), and the activities of protective enzymes (including SOD, POD, and CAT) in the leaves were significantly increased, while the MDA content in the leaves was decreased. Conclusion: Soaking seeds with 0.05-0.60 mmol/L SNP could promote the salt resistance of the seeds and seedlings of S. marianum. The different cultivars of S. marianum differ in the sensitivity to SNP. The optimal concentration of SNP for the seed soaking of S. marianum with white and black skins is 0.10 and 0.40 mmol/L, respectively.

Silybin-derived flavonolignans and their structure-activity relationships: Research advances / 国际药学研究杂志

Silybum marianum (L.) Gaertn. is recorded by the pharmacopoeias of many countries to have such bioactivities as antioxidant, anti-hepatocirrhosis, anti-inflammation, immunomodulation and liver regeneration. It has been used clinically to treat hepatitis, fatty liver, liver cirrhosis, ischemic damage, radiation injury, etc. With silybin entering the phase II clinical trial as an anti-cancer drug, its correlative investigations including derivatives preparation, structure-activity relationship (SAR) detection, and the detailed action mechanisms have been developing rapidly. Recently, the new synthetic strategy of silybin and dehydrosilybin (DHS) along with their anti-HSV (herpes simple virus) activity has also been reported. This paper reviews about one hundred preparative derivatives of silybin and DHS which appeared in recent years, with the emphasis on their pharmacological activity and the results of their SAR. The future development direction of structural modification of silybin and DHS is also discussed and predicted.