Genetic activity and cytotoxic effect of Silybum marianum extract in comparison with the impact of antitumor drugs etoposide and methotrexate: a study on Drosophila melanogaster.

One of the most important directions in medical practice involves the exploration of protectors and adaptogens to mitigate side effects associated with the use of antitumor and potent medications. The extracts of medicinal plants are actively tested as protectors. In connection with this, the aim of the research was to determine the effectiveness of the protective properties of Silybum marianum extract against the drugs 'etoposide' and 'methotrexate' at concentrations of 800 µg/kg using the model object Drosophila melanogaster. To analyse the protective properties, the following research methods were applied: determining the lethal concentration, average individual fertility of individuals, mortality of F1 offspring at embryonic stages, SMART (a method for determining the frequency of somatic mutations and recombinations) and assessing the intensity of programmed cell death. The study revealed that a 5% S. marianum extract reduces the genotoxicity of etoposide and exhibits a high cytotoxic effect, both independently and in combination with etoposide and methotrexate.

Application of SNAP-tag-EGFR cell membrane chromatography model in screening antitumor active components of Silybum marianum (L.) Gaertn.

The SNAP-tag-epidermal growth factor receptor (SNAP-tag-EGFR) cell membrane chromatography (CMC) model is a powerful tool for investigating ligand-receptor interactions and screening active ingredients in traditional Chinese medicine. Most tyrosine kinase inhibitors (TKIs) target epidermal growth factor receptors. However, TKIs associated with significant side effects and drug resistance must be addressed immediately. Therefore, there is an urgent need to develop new TKIs with high efficiency and low toxicity. Because of its low toxicity and side effects, traditional Chinese medicine has been widely employed to treat various diseases, including cancer. Hence, this study aimed to use the SNAP-tag-EGFR/CMC-high-performance liquid chromatography-mass spectrometry (HPLC-MS) two-dimensional system model as the research tool to screen and identify potential EGFR antagonists from the Chinese medicine Silybum marianum (L.) Gaertn. The applicability of the system was verified using the positive control drug osimertinib. Four potential EGFR antagonists were screened from the Chinese medicine Silybum marianum (L.) Gaertn.. They were identified as silydianin, silychristin, silybin, and isosilybin. Additionally, their pharmacological activity was preliminarily verified using a CCK-8 assay. The kinetic parameters of the four active ingredients interacting with EGFR and their binding modes with EGFR were analyzed using nonlinear chromatography (NLC) and molecular docking. This study identified silydianin, silychristin, silybin, and isosilybin from Silybum marianum (L.) Gaertn. and verified their potential antitumor effects on EGFR.

A Genomic Evaluation of Six Selected Inbred Lines of the Naturalized Plants of Milk Thistle (Silybum marianum L. Gaertn.) in Korea.

Milk thistle (Silybum marianum) belongs to the Asteraceae family and is a medicinal plant native to the Mediterranean Basin. Silymarin in achene is a widely used herbal product for chronic liver disease. There is growing interest in natural medicine using milk thistle in Korea, but the raw material completely relies on imports. Despite its economic importance, phenotypic evaluations of native resources of milk thistle in Korea have not been carried out. In addition, genomic research and molecular marker development are very limited in milk thistle. In this study, we evaluated 220 milk thistle resources consisting of 172 accessions collected from the domestic market, and 48 accessions isolated from 6 accessions distributed by the National Agrobiodiversity Center in Korea. Six plant characteristics (height, seed weight, number of flowers, seed weight per flower, spine length, and color at harvest) were measured, and six samples (M01-M06) were selected to represent the genetic diversity of the population for genomic research. To develop PCR-based and co-dominant insertion/deletion (InDel) markers, we performed genome-wide InDel detection by comparing the whole-genome resequencing data of the six selected accessions with the reference genome sequence (GCA_001541825). As a result, 177 InDel markers with high distinguishability and reproducibility were selected from the 30,845 InDel variants. Unknowingly imported alien plant resources could easily be genetically mixed, and jeopardized seed purity can cause continuous difficulties in the development of high value-added agricultural platforms utilizing natural products. The selected plant materials and 177 validated InDel markers developed via whole-genome resequencing analysis could be valuable resources for breeding, conservation, and ecological studies of natives to Korea, along with acceleration of Silybum marianum industrialization.

An Expansion of the Endoplasmic Reticulum that Halts Autophagy is Permissive to Genome Instability.

The links between autophagy and genome stability, and whether they are important for lifespan and health, are not fully understood. We undertook a study to explore this notion at the molecular level using Saccharomyces cerevisiae. On the one hand, we triggered autophagy using rapamycin, to which we exposed mutants defective in preserving genome integrity, then assessed their viability, their ability to induce autophagy and the link between these two parameters. On the other hand, we searched for molecules derived from plant extracts known to have powerful benefits on human health to try to rescue the negative effects rapamycin had against some of these mutants. We uncover that autophagy execution is lethal for mutants unable to repair DNA double strand breaks, while the extract from Silybum marianum seeds induces an expansion of the endoplasmic reticulum (ER) that blocks autophagy and protects them. Our data uncover a connection between genome integrity and homeostasis of the ER whereby ER stress-like scenarios render cells tolerant to sub-optimal genome integrity conditions.

Combinatorial treatment with Silybum marianum essential oil enhances the therapeutic efficacy of a 5-fluorouracil base therapy for hepatocellular carcinoma.

Silybum marianum seeds contain a family of flavonolignans which can regulate cancer cell proliferation and apoptosis. However, research has rarely focused on the effect of S. marianum essential oil in combination with another anticancer drug. Here, we evaluated the antitumor effect of a combination of 5-fluorouracil (5-FU) and S. marianum essential oils on some pathways in hepatocellular carcinoma (HCC) in vitro and in vivo. Gas chromatography-mass spectrography results indicated there was no significant difference between the components of essential oils isolated from two geographical areas (Khuzestan or Isfahan, Sm-K or Sm-I). Each preparation decreased the viability of H22 cells compared to the control group. S. marianum essential oils alone, and combined with 5-FU, reduced the migration and invasion of H22 cells. Angiogenesis-related proteins were significantly reduced both in vivo and in vitro. Apoptosis and autophagy-related proteins were modulated both in vivo and in vitro. Each treatment decreased phospho-NF-κB (p65) and NF-κB (p65) protein levels. Expression levels of Wnt pathway-related genes were also decreased both in vivo and in vitro. Results revealed that the combination of S. marianum and 5-FU prolonged survival in a mouse model of HCC compared to either treatment alone.

Modulation of Insulin Resistance by Silybum marianum Leaves, and its Synergistic Efficacy with Gymnema sylvestre, Momordica charantia, Trigonella-foenum graecum Against Protein Tyrosine Phosphatase 1B.

Prolonged insulin resistance is considered one of the reasons for Type 2 Diabetes Mellitus. Upregulation of Protein tyrosine phosphatase 1B (PTP1B), a negative regulator of insulin signalling, has been well studied as a key regulator in prognosis to insulin resistance. It has been widely studied as a desirable molecular therapeutic target. The study aimed to evaluate the efficacy of leaf extract of the medicinal plants Silybum marianum on the inhibition of PTP1B activity. It also explored the synergistic effect with extracts of Gymnema sylvestre (leaves), Momordica charantia (seeds), and Trigonella foenum graecum (seeds). The S. marianum leaves showed dose-dependent inhibition of PTP1B ranging from 9.48-47.95% (25-1000 µg mL-1). Assay with individual plant extracts showed comparatively lesser inhibition of PTP1B as compared to metformin as a control (38% inhibition). However, a synergistic effect showed nearly 45% PTP1B inhibition (higher than metformin) after the assay was done with selected four plant extracts in combination. The effect of leaf extracts of S. marianum was studied for glucose uptake efficiency in yeast cell lines which was found to be increased by 23% as compared to the control (without extract). Metformin improves glucose upake by yeast cells by ~15-31%. GC-MS analysis revealed 23 phytochemicals, some of which possessed anti-diabetic properties. A dose-dependent increase in antioxidant activity of S. marianum leaves extracts was observed (40-53%). The findings of the study highlighted the presence of various phytochemicals in leaves extracts that are effective against PTP1B inhibition and may help in reinvigorating drug development.

A review of therapeutic potentials of milk thistle (Silybum marianum L.) and its main constituent, silymarin, on cancer, and their related patents.

For more than 2000 years, Silybum marianum L. (milk thistle) has been used for treating different complications such as jaundice, hepatitis, and cancers. It has also been shown that silymarin, a flavonolignan extract of the plant, demonstrates chemopreventive effects against cancers. This patent review presents and discusses recent patents concerning the anticancer effects of S. marianum and silymarin. The data were gathered by searching an extensive literature review conducted in Google Scholar, PubMed, Scopus, Google Patent, Patent Scope, and US Patent. Milk thistle and silymarin have been used in a variety of medical, therapeutic, and pharmaceutical fields, according to a large number of documents and patents. Milk thistle and silymarin have been used as complementary treatments for cancers such as skin, prostate, and colorectal cancers, as well as hepatoprotective agents. Silymarin exerts a chemopreventive effect on reactivating cell death pathways by modulation of the antiapoptotic proteins and synergizing with agonists of death domain receptors. Based on the results of these patents, silymarin could be beneficial to oncology patients, especially for the treatment of the side effects of anticancer chemotherapeutics. Following the human propensity to use phytocompounds rather than medicines based on chemical constituents, special attention must be paid to tie the value of milk thistle and silymarin from basic science to clinical applications.

Investigation of market herbal products regulated under different categories: How can HPTLC help to detect quality problems?

Background: Herbal products regulated under different categories were found to be of different quality. This has been demonstrated by the increasing number of reports on the quality of herbal products in the scientific literature. Proper identification is an effective way to address this concerning issue early on in a products' manufacturing process. Objectives: To assess the quality of milk thistle, coneflower and black cohosh herbal drugs, preparations and products commercialized under different regulatory categories, and to illustrate the usefulness of HPTLC as a tool for evaluating quality. Methods: HPTLC methods were adapted from the European Pharmacopeia's monographs for milk thistle fruits, black cohosh and purple coneflower. Additional detection modes beyond those described in the monographs were employed, and the entire HPTLC fingerprints were used for examination of identity and purity of the investigated samples. Results: All products regulated as Traditional Herbal Medicinal Products were shown to be of high quality: their fingerprints were consistent and without unexpected zones. A significant number of food supplements show quality issues (mainly adulterations): 52.4% of milk thistle, 33.3% of coneflower, and 45.5% of black cohosh products. The same was observed in 66.6% of black cohosh herbal drugs and preparations.

Hydroponics and elicitation, a combined approach to enhance the production of designer secondary medicinal metabolites in Silybum marianum.

Medicinal plants have been used to cure human diseases since decades. Silybum marianum, a medicinal plant, is regarded as a source of secondary metabolites with therapeutic value against liver diseases and diabetes. The present study was conducted to enrich the production of secondary metabolites in the vegetative parts of Silybum marianum using elicitation strategy in hydroponic system with different elicitors. The elicitors of fungus Aspergillus niger (0.2 g/L), methyl jasmonate (MeJA) (100 µM) and silver nanoparticles (AgNPs) (1 ppm) were added in hydroponic medium, individually and in combination form to the 15 days old plant. The elicitor-treated plants were harvested at different time points (24-144 h; increment 24 h) and their biochemical parameters like phenolics, flavonoids, nitric oxide (NO), and superoxide dismutase (SOD) were analyzed. The results showed hyper-accumulation of these biochemical contents, especially in response to MeJA (100 µM), followed by AgNPs (1 ppm) and co-treatment of AgNPs (1 ppm) with other elicitors. The results revealed that the treatment with MeJA (100 µM) exhibited the highest flavonoid (304 µg g-1), phenolic (372 µg g-1), and SOD (16.2 U g-1) contents. For NO levels, the maximum value of 198.6 nmole g-1 was achieved in response to the treatment with MeJA + Green synthesized AgNPs (100 µM + 1 ppm). Our findings depicted an enhanced production of medicinally important plant secondary metabolites and antioxidants; hence, the method applied in this study can play a significant role to improve therapeutic values of the plants.

One-step sustainable extraction of Silymarin compounds of wild Algerian milk thistle (Silybum marianum) seeds using Gas Expanded Liquids.

This work reports the application of Gas Expanded Liquid (GXL) extraction to concentrate the flavonolignan fraction (silymarin) and taxifolin from Silybum marianum seeds, which have proven to be highly valuable health-promoting compounds. GXL using green solvents was used to isolate silymarin with the objective of replacing conventional methods. In one hand, the effect of different compositions of solvents, aqueous ethanol (20%, 50% or 80% (v/v)) at different CO2/liquid (25, 50 and 75%) ratios, on the GXL extraction was investigated. The obtained extracts have been chemically and functionally characterized by means of UHPLC-ESI-MS/MS (triple quadrupole) and in-vitro assays such as anti-inflammatory, anti-cholinergic and antioxidant. Results revealed that the operating conditions influenced the extraction yield, the total phenolic content and the presence of the target compounds. The best obtained yield was 55.97% using a ternary mixture of solvents composed of CO2:EtOH:H2O (25:60:15) at 40 °C and 9 MPa in 160 min. Furthermore, the results showed that obtained extracts had significant antioxidant and anti-inflammatory activities (with best IC50 value of 8.80 µg/mL and 28.52 µg/mL, respectively) but a moderate anti-cholinesterase activity (with best IC50 value of 125.09 µg/mL). Otherwise, the concentration of silymarin compounds in extract can go up to 59.6% using the present one-step extraction method without further purification, being silybinA+B the predominant identified compound, achieving value of 545.73 (mg silymarin/g of extract). The obtained results demonstrate the exceptional potential of GXL to extract high-added values molecules under sustainable conditions from different matrices.

Effect of herbal cream containing Fumaria officinalis and silymarin for treatment of eczema: A randomized double-blind controlled clinical trial.

Objective: Atopic dermatitis (AD) is a common skin disorder with symptoms including severe pruritus and eczematous lesions. AD affects between 5 and 20% of people in their life. Silymarin (SM) is a polyphenolic flavonoid from Silybum marianum L. and has several therapeutic characteristics including antiallergic, anticancer, and anti-inflammatory properties. Fumaria officinalis is a small plant that has a high antioxidant power and modulating effects on the immune system. Therefore, the current study intended to examine the influence of these two herbs extract on severity and symptoms of AD in patients. Materials and Methods: 40 patients with mild to moderate eczema randomly received mometasone 0.1% or the herbal cream. Treatment course was 2 weeks and patients were examined before and after 2 weeks of treatment using the SCORAD system. Results: The reduction of SCORAD score was significant in both groups (p=0.04 in the herbal group and p=0.03 in the mometasone group) but no significant difference was observed between the groups. Mean SCORAD score was 27.66±5.9 before therapy and 4.77±1.6 after therapy in the mometasone group and mean SCORAD score was 26.05±7.1 before therapy and 6.944±2.6 after therapy in the herbal group. Conclusion: The current study indicated the impact of these two herbs extract on severity and symptoms of AD in patients; these plants may be a new treatment in reducing eczema symptoms and its problems.

Effect of milk thistle (Silybum marianum) supplementation on the serum levels of oxidative stress markers in male half marathon athletes.

CONTEXT: Increased aerobic metabolism during exercise is a potential source of oxidative stress and the use of herbal medicines as a dietary supplement rich in antioxidants is an interesting and controversial concept that have been considered during the past decades. Objective: The purpose of the present study was to investigate the effects of Silybum marianum (SM) on exercise-induced oxidative stress in half marathon athletes. MATERIALS AND METHODS: Phytochemical Analysis in aqueous extract of SM leaves and seeds were determined. Forty healthy male athletes were divided into four groups (n = 10): control group(G1), G2 supplemented with 100 mg of SM leaves/kg/day, G3 supplemented with 100 mg of SM seeds/kg/day, and G4 supplemented with 100 mg of SM leaves + seeds/kg/day. The effects of SM on malondialdehyde (MDA) and antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH)] were assessed. RESULTS: Aqueous extract of SM leaves have good DPPH free radical scavenging activity and the highest content of total polyphenols. A significant increase of serum SOD, CAT, and GSH levels and reduction in the levels of MDA in the serum of athletes supplemented with aqueous extract of seeds and leaves of SM was detected. CONCLUSION: SM supplement offered protection against exercise-induced oxidative stress.

Hepatoprotective mechanism of Silybum marianum on nonalcoholic fatty liver disease based on network pharmacology and experimental verification.

The study aimed to identify the key active components in Silybum marianum (S. marianum) and determine how they protect against nonalcoholic fatty liver disease (NAFLD). TCMSP, DisGeNET, UniProt databases, and Venny 2.1 software were used to identify 11 primary active components, 92 candidate gene targets, and 30 core hepatoprotective gene targets in this investigation, respectively. The PPI network was built using a string database and Cytoscape 3.7.2. The KEGG pathway and GO biological process enrichment, biological annotation, as well as the identified hepatoprotective core gene targets were analyzed using the Metascape database. The effect of silymarin on NAFLD was determined using H&E on pathological alterations in liver tissues. The levels of liver function were assessed using biochemical tests. Western blot experiments were used to observe the proteins that were expressed in the associated signaling pathways on the hepatoprotective effect, which the previous network pharmacology predicted. According to the KEGG enrichment study, there are 35 hepatoprotective signaling pathways. GO enrichment analysis revealed that 61 biological processes related to the hepatoprotective effect of S. marianum were identified, which mainly involved in response to regulation of biological process and immune system process. Silymarin was the major ingredient derived from S. marianum, which exhibited the hepatoprotective effect by reducing the levels of ALT, AST, TC, TG, HDL-C, LDL-C, decreasing protein expressions of IL-6, MAPK1, Caspase 3, p53, VEGFA, increasing protein expression of AKT1. The present study provided new sights and a possible explanation for the molecular mechanisms of S. marianum against NAFLD.

System biology-based investigation of Silymarin to trace hepatoprotective effect.

Silymarin is used as a hepatoprotective agent since ancient times which could be via its potent anti-oxidant effect. However, the mode of silymarin for the hepatoprotective effect has not been established with the targets involved in hepatic cirrhosis. The present study investigated the multiple interactions of the flavonolignans from Silybum marianum with targets involved in hepatic cirrhosis using a series of system biology approaches. Chemo-informative tools and databases i.e. DIGEP-Pred and DisGeNET were used to predict the targets of flavonolignans and proteins involved in liver cirrhosis respectively. Further, STRING was used to enrich the protein-protein interaction for the flavonolignans-modulated targets. Similarly, molecular docking was performed using AutoDock Vina. Additionally, molecular dynamics simulation and MM-PBSA calculations were carried out for the lead-hit complexes by GROMACS. Thirteen flavonolignans were identified from S. marianum, in which silymonin exhibited the highest drug-likeness score i.e. 1.09. Similarly, CTNNB1 was found to be regulated by the 12 different flavonolignans and was majorly expressed within the compound(s)-protein(s)-pathway(s) network. Further, silymonin had the highest binding affinity; binding energy -9.2 kcal/mol with the CTNNB1 and formed very stable hydrogen bond interactions with Arg332, Ser336, Lys371, and Arg475 throughout 100 ns molecular dynamic production run. The binding free energy of CTNNB1-silymonin complex was found to be -15.83 ± 2.71 kcal/mol. The hepatoprotective property of S. marianum may be due to the presence of silymonin and silychristin; this could majorly modulate CTNNB1, HMOX1, and CASP8 in combination with other flavonolignans. Our findings further suggest designing the in-vitro and in-vivo studies to validate the interaction of flavonolignans with identified targets to strengthen present findings of S. marianum as a hepatoprotective..

Novel nutraceutical supplements with yeast ß-glucan, prebiotics, minerals, and Silybum marianum (silymarin) ameliorate obesity-related metabolic and clinical parameters: A double-blind randomized trial.

Purpose: It is known that obesity has a multifactorial etiology that involves genetic and environmental factors. The WHO estimates the worldwide prevalence of 1.9 billion overweight adults and more than 650 million people with obesity. These alarming data highlight the high and growing prevalence of obesity and represent a risk factor for the development and aggravation of other chronic diseases, such as nonalcoholic fatty liver disease (NAFLD) that is frequently considered the hepatic outcome of type 2 diabetes. The use of non-pharmacological therapies such as food supplements, nutraceuticals, and natural integrative therapies has grown as an alternative tool for obesity-related diseases compared to conventional medications. However, it is a still little explored research field and lacks scientific evidence of therapeutic effectiveness. Considering this, the aim is to evaluate whether a new nutraceutical supplement composition can improve and supply essential mineral nutrients, providing an improvement of obesity-related metabolic and endocrine parameters. Methods: Sedentary volunteers (women and men) with body mass index (BMI) ≤34.9 kg/m2 were divided into two groups: Novel Nutraceutical Supplement_(S) (n = 30) and Novel Nutraceutical Supplement (n = 29), differing in the absence (S) or presence of silymarin, respectively. Volunteers were instructed to take two capsules in the morning and two capsules in the evening. No nutritional intervention was performed during the study period. The data (anthropometrics and anamneses) and harvest blood (biochemistry and hormonal exams) were collected at three different time points: baseline time [day 0 (T0)], day 90 (T90), and day 180 (T180) post-supplementation. Results: In the anthropometric analysis, the waist circumference in middle abdomen (WC-mid) and waist circumference in iliac crest (WC-IC) were reduced. Also, the waist-to-height ratio (WHt R) and waist-to-hip ratio (WHR) seem to slightly decrease alongside the supplementation period with both nutraceutical supplements tested as well as transaminase enzyme ratio [aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio (AAR)], a known as a biomarker of NAFLD, and endocrine hormones cortisol and thyroid-stimulating hormone (TSH) at 90 and 180 days post-supplementation. Conclusions: In a condition associated with sedentary and no nutritional intervention, the new nutraceutical supplement composition demonstrated the ability to be a strong and newfangled tool to improve important biomarkers associated with obesity and its comorbidities.

Silybum marianum total extract, silymarin and silibinin abate hepatocarcinogenesis and hepatocellular carcinoma growth via modulation of the HGF/c-Met, Wnt/ß-catenin, and PI3K/Akt/mTOR signaling pathways.

Hepatocellular carcinoma (HCC) has been identified as one of the most deadly malignancies with limited therapeutic efficacy worldwide. However, understanding the molecular mechanisms of crosstalk between signaling pathways in HCC and predicting cancer cell responses to targeted therapeutic interventions remain to be challenge. Thus, in this study, we aimed to evaluate the anticancerous efficacy of Silybum marianum total extract (STE), silymarin (Sm), and silibinin (Sb) against experimentally-induced HCC in rats. In vitro investigations were also performed and the anticancer effects against HCC cell lines (HepG2 and Huh7) were confirmed. Wistar rats were given diethylnitrosamine (DEN)/2-acetylaminofluorene (AAF)/carbon tetrachloride (CCl4) and were orally treated with STE (200 mg/kg body weight (bw)), Sm (150 mg/kg bw), and Sb (5 mg/kg bw) every other day from the 1st or 16th week to the 25th week of DEN/AAF/CCl4 injection. Treatment with STE, Sm, and Sb inhibited the growth of cancerous lesions in DEN/AAF/CCl4-treated rats. This inhibition was associated with inhibition of Ki-67 expression and repression of HGF/cMet, Wnt/ß-catenin, and PI3K/Akt/mTOR signaling pathways. STE, Sm, and Sb improved liver function biomarkers and tumor markers (AFP, CEA, and CA19.9) and increased total protein and albumin levels in serum. STE, Sm, and Sb treatment was also noted to reduce the hepatic production of lipid peroxides, increase hepatic glutathione content, and induce the activities of hepatic antioxidant enzymes in DEN/AAF/CCl4-treated rats. These results indicate that STE, Sm, and Sb exert anti-HCC effects through multiple pathways, including suppression of Ki-67 expression and HGF/cMet, Wnt/ß-catenin, and PI3K/Akt/mTOR pathways and enhancement of antioxidant defense mechanisms.

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.

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

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.

Evaluation the effect of Silybum marianum ointment on episiotomy wound healing and pain intensity in primiparous women: a randomized triple blind clinical trial.

BACKGROUND: Episiotomy is the most commonn surgical procedure in midwifery which as any other wounds can cause infection or delay in healing. The current study aimed to determine effect of Silybum marianum ointment on pain severity and healing of episiotomy wound in primiparous women referring to Shahid Nourani Hospital at 2019. METHODS: This research was done as a randomized, triple-blind clinical trial on 87 priiparous women (44 indivdiuals in Silybum marianum ointment group and 43 indivdiuals in placebo group) referred to Shahid Nourani Hospital in Talesh (Guilan Province), Iran at September 2019. After labor and performing episiotomy, twice a day for 10 days as a fingertip size of the ointment was prescribed to be topically used on the episiotomy incision for both groups (Silybum marianum ointment or placebo ointment). Data gathering was done using demographic and midwifery information questionnaire, Episiotomy healing assessment: Redness, Edema, Ecchymosis, Discharge, Approximation)REEDA Scale (REEDA Scale: Redness(R); Edema (E), Ecchymosis(E), Discharge from the wound(D); Approximation of the perineal tissues(A))(scale, and visual analogue scale of pain. Examination of healing status of the perinea incision was performed during first 12 h, fifth day and tenth day after labor.Kolmogrov-Smirnov test was used in order to investiagte nomrality of data distribution of quantitative data, and two- independent samples t test, Chi square, repeated measures two factorial analysis of variance and Fischer's exact test were used. SPSS software version 23 was used to analyze data and 0.05 was considered as signifcance level. RESULTS: Both groups of Silybum marianum and placebo groups did not differ regarding demographic and midwifery characteristics, hygiene status prior to intervention (P > 0.05). Mean difference of pain severity and REEDA scale in Silybum marianum ointment group in 12 h after labor, at fifth day and tenth day after labor was significant comparing to control group which indicates decline in episiotomy pain severity and faster wound healing (P < 0.001). CONCLUSIONS: Silybum marianum ointment ointment accelerates episiotomy wound healing rate due to its healing properties and decreases pain severity. TRIAL REGISTRATION: This study was registered in Iranian Registry of Clinical Trials in 10/08/2019 with the IRCT ID: IRCT201811100411603N1.