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Silymarin, a flavonolignan from the seeds of 'milk thistle' (Silybum marianum), has been widely used from ancient times because of its excellent hepatoprotective action. It is a mixture of mainly three flavonolignans, viz, silybin, silidianin, and silychristine, with silybin being the most active. Silymarin has been used medicinally to treat liver disorders, including acute and chronic viral hepatitis, toxin/drug-induced hepatitis, and cirrhosis and alcoholic liver diseases. It has also been reported to be effective in certain cancers. Its mechanism of action includes inhibition of hepatotoxin binding to receptor sites on the hepatocyte membrane; reduction of glutathione oxidation to enhance its level in the liver and intestine; antioxidant activity; and stimulation of ribosomal RNA polymerase and subsequent protein synthesis, leading to enhanced hepatocyte regeneration. It is orally absorbed but has very poor bioavailability due to its poor water solubility. This review focuses on the various pharmacological activities of silymarin including the clinical trials. For the first time, the review also looks at the formulation work that has been done to enhance its solubility, so as to increase its bioavailability and thus, its hepatoprotective action.
Various experimental studies using compounds that directly or indirectly cause liver damage have been carried out to demonstrate the hepatoprotective action of silymarin in xenobiotic intoxication and fungal intoxication.
Amanita phalloids toxin :Administration of this compound produces acute intoxication in mice /rats/rabbits/dogs. Silymarin at 50-150 mg/kg intravenous dose provided protection and cure.  The increase in liver enzymes and reduction in coagulation factor seen with sublethal doses of A. phalloids can be prevented with silymarin.  This is evident at 5 h with silymarin given intravenously at a dose of 50 mg/kg and at 24 h with a dose of 30 mg/kg.
Gupta et al ., have shown the anti-inflammatory and antiarthritic activity of silymarin, which is mediated through the inhibition of 5-lipoxygenase.  The anti-inflammatory effect was tested on carrageenan-induced oedema, papaya latex-induced oedema, and arachidonic acid (AA)-induced mouse ear oedema. The results indicated that the activity was less marked in the carageenan model but significant in the papaya latex and AA models of inflammation.
In the AA model, a 25 mg/kg P.O. dose of silymarin produced 36.84% inhibition of oedema, which the authors attributed to its inhibitory action on the formation of 5-lipoxygenase and the leukotrienes involved in inflammation.
Antiarthritic activity was tested in mycobacterial adjuvant-induced arthritis in rats. When tested at 6.25, 12.5, and 25 mg/kg P.O. dose, silymarin showed a dose-related inhibition of 14.87, 23.73, and 31.64% on day 13, while boswellic acids employed as the positive control showed 32.91% inhibition. This showed that the silymarin was more effective in cases of developing arthritis compared to developed arthritis.
Škottovaa et al ., investigated currant oil (from Ribes nigrum L.)-induced modulation of the antihypercholesterolemic and LDL antioxidant effects of silymarin and (the better bioavailable) silibinin-phosphatidylcholine complex (SPC) in rats fed on a high-cholesterol, high-fat diet.  They fed rats on a high-cholesterol diet supplemented with 10% of currant oil containing polyunsaturated fatty acids (PUFA, 61.1% of n-6 and 15.4% of n-3) and lower amounts of saturated (SFA, 7.7%) and monounsaturated (MUFA, 14.3%) fatty acids which caused a significant lowering of plasma cholesterol associated with a mild decrease in VLDL-C and an increase in HDL-C, when compared to rats fed on high-cholesterol diet with 10% of lard fat containing low amounts of PUFA (7.7% of n-6 and 0.7% of n-3) and higher amounts of SFA (42.7%) and MUFA (47.5%). However, currant oil feeding led to the increased oxidizability of LDL. It was found that silymarin, but not SPC, was effective in preventing the development of dietary-induced hypercholesterolemia in both dietary fats, with a slightly better result in rats fed the diet containing currant oil. On the other hand, SPC was more effective than silymarin in suppressing LDL oxidizability. The results suggest that the antihypercholesterolemic effect of silymarin in rats fed on a high-cholesterol diet is improved by dietary currant oil, but the currant oil induces an increased oxidizability of LDL. This can be suppressed by improvement of the bioavailability of silibinin, as demonstrated here with the silibinin-phosphatidylcholine complex.
Lirussi et al ., investigated silybin-β-cyclodextrin complex in the treatment of patients with diabetes mellitus and alcoholic liver disease. In noninsulin dependent diabetes mellitus (type 2) and associated chronic liver disease, plasma levels of glucose, insulin, and triglycerides are high, with increased lipid peroxidation and reduction in natural antioxidant reserves. It was hypothesized by the authors that a better glucose and lipid metabolism could be achieved by the rebalancing of cell redox levels and amelioration of liver function. They assessed the effect of the silybin-β-cyclodextrin formulation in 60 patients with chronic alcoholic liver disease and concomitant type 2 diabetes mellitus in a double-blind study for 6 months.  The parameters measured included fasting and mean daily plasma glucose levels, glycosylated hemoglobin (HbA 1c ), basal and stimulated C-peptide and insulin levels, and HDL-cholesterol and triglycerides levels, in addition to conventional liver function tests. Insulin sensitivity was estimated by HOMA-IR. Malondialdehyde (MDA) was also measured before and after treatment as an index of oxidative stress. The results showed that the oral administration of silybin-β-cyclodextrin in patients with type 2 diabetes and compensated chronic alcoholic liver disease caused a significant decrease in both glucose and triglyceride plasma levels. These effects were attributed to the recovery of energy substrates with an improved insulin activity and a reduced lipid peroxidation.
Kim et al ., studied the comparative bioavailability of Liverman capsule to Legalon capsule (each containing silymarin) and silymarin tablet in 24 healthy Korean volunteers.  Each volunteer received each medicine at the silibinin dose of 120 mg in a 3 × 3 crossover study, with a 1-week washout period between the doses. Plasma concentrations of silibinin were monitored by HPLC for a period of 12 h after the administration. AUC 0-∞, C max , and t max were obtained from the plasma concentration-time data. After oral administration of each medicine, the pharmacokinetic parameters of silibinin, viz, AUC 0-12h , AUC 0-∞, C max , and t max were obtained as follows: Legalon capsule (5.59 mg/ml × h, 6.00 mg/ml × h, 1.33 mg/ml, and 1.83 h), silymarin tablet (4.24 mg/ml × h, 4.63 mg/ml × h, 1.13 mg/ml and 2.10 h) and Liverman capsule (13.9 mg/ml × h, 15.1 mg/ml × h, 6.04 mg/ml and 0.875 h). This showed that all the pharmacokinetic parameters were significantly greater for Liverman capsule compared to Legalon capsule and silymarin tablet, indicating that the absorption and the extent of relative oral bioavailability of silibinin after Liverman capsule were significantly faster and greater than that after Legalon capsule and silymarin tablet, respectively.
In another study designed to assess the absorption of silymarin bound to phosphatidylcholine, plasma silybin levels were determined after administration of single oral doses of silymarin-phosphatidylcholine complex and a similar amount of silymarin to nine healthy volunteers.  The bioavailability of the silymarin-phosphatidylcholine complex was much greater than that of silymarin in spite of the rapid absorption with both preparations, as indicated by the higher plasma silybin levels at all sampling times after intake of the complex. The authors concluded that complexation with phosphatidylcholine greatly increases the bioavailability of silymarin, probably by facilitating its passage across the gastrointestinal mucosa.
Vailati et al ., in their study designed primarily to evaluate the dose-response relationship of phosphatidylcholine-bound silymarin, demonstrated the positive effects of the complex.  In their study, patients with virus- or alcohol-induced chronic hepatitis were given different doses of the complex: twenty patients received 80 mg twice daily, twenty received 120 mg twice daily, and another twenty patients received 120 mg three times daily, for 2 weeks. At all the doses, phosphatidylcholine-bound silymarin produced a statistically significant decrease of mean serum and total bilirubin levels. When used at dosage of 240 or 360 mg per day, similar results were observed with ALT and GGTP liver enzymes. These results indicated the effectiveness of relatively low doses of phosphatidylcholine-bound silymarin in the short-term treatment of hepatitis caused by viruses or alcohol, though the best results are achieved at higher doses.
Carducci et al ., in a case study, reported that silybin hemisuccinate given by the intravenous route had a favourable effect on a family of four suffering from severe liver damage caused by Amanita mushroom (amatoxin) compared to when they were treated with the standard therapy.  Moreover, investigations done after 2 months showed an absence of any morphological alteration in the hepatobiliopancreatic echography, suggesting that silybin may play a significant role in hepatic tissue protection.
The therapeutic indications mentioned above clearly indicates the potential of silymarin as a natural immunomodulator, but the problem with the use of silymarin lies in its poor bioavailability. Due to this reason the dose of silymarin given needs to be large so as to achieve therapeutic plasma levels. Various groups of scientists have used different approaches to address this problem and have succeeded in improving the bioavailability of silymarin.
In order to improve the dissolution rate, Soo Woo et al . formulated silymarin in the form of a self-microemulsifying drug-delivery system (SMEDDS).  Based on pseudo-ternary phase diagram, the optimum formulation was obtained, consisting of 15% silymarin, 10% glyceryl monooleate as the oil phase, 37.5% mixture of polysorbate 20 and HCO-50 (1:1) as the surfactant, and 37.5% transcutol as the cosurfactant, with a surfactant/cosurfactant ratio of 1. The mean droplet size of the oil phase in the microemulsion was 67 nm. The release profile of silybin from the prepared SMEDDS capsule showed a significant increase in drug release over the reference capsule (Legalonβ). The percentage release of silybin from SMEDDS was 2.5 times higher than reference after 6 h. The pharmacokinetic evaluation of silymarin showed that there was an approximately 2-fold decrease in the t max from SMEDDS, 8-fold enhancement in the C max , and 3.6 times increase in the AUC, showing that there was a significant increase in the bioavailability of silymarin from SMEDDS over the conventional reference dosage form.
Yanyu et al . prepared silybin-phospholipid complexes to increase the bioavailability of orally administered silymarin and compared the pharmacokinetic characteristics and bioavailability after administration of silybin-phospholipid complex and silybin-N-methylglucamine in rats.  With ethanol as the reaction medium, silybin and phospholipid were dissolved in the organic medium and the solvent was removed under vacuum, which resulted in the formation of the complex. The solubility studies showed a 1000-fold increase in the solubility of the silybin complex in water and octanol compared to the solubility of the physical mixture. Dissolution of silybin complex at pH 6.8 was significantly more than at pH 1.2. At the end of 60 min the amount of silybin dissolved was 158.4 mg at pH 6.8 compared to 6.3 mg at pH 1.2. It was found that the rise in pH increased the dissolution of silybin. Rat bioavailability studies showed more than 5-fold enhancement in the bioavailability of silymarin from the phospholipid complex than the silybin-N-methylglucamine.
Yanyu et al . prepared silymarin proliposomes to increase the bioavailability of oral silymarin in beagle dogs.  The proliposomes were prepared by film-deposition of a silymarin and phospholipid mixture on a mannitol carrier in a round-bottom flask. Dissolution of proliposomes at pH 1.2 and 6.8 showed that the dissolution of silymarin was complete after 20 min, irrespective of the pH of the media. The dissolution of silymarin from the proliposomes was more than that from the control (silymarin-loaded mannitol powder prepared by same method as the proliposomes, but without the phospholipds). The encapsulation efficacy of the formulation was more than 90%, had a mean particle size of 196.4 nm, and was stable for 3 months at 40°C. The pharmacokinetic parameters for silymarin proliposomes and silymarin showed a t max of 30 min for both and C max of 472.62 and 89.78 ng/ml, and AUC of 2606.21 and 697 ng/ml, respectively, demonstrating enhanced GI absorption of silymarin from the proliposomes.
El-Samaligy et al . prepared the silymarin hybrid liposomes by reverse evaporation technique, using lecithin, cholesterol, stearyl amine, and tween 20 in a molar ratio of 9:1:1:0.5.  The prepared liposomes showed an encapsulation efficiency of 69.22%. Mixing silymarin-loaded liposomes with unloaded ones in 1:1 proportion was useful in the prevention of aggregates, which threaten liposomes stability. This selected formulation was stable with respect to encapsulation efficiency and particle size after 3 months of storage at 4°C. Differential scanning calorimetery and Fourier Transform Infrared spectroscopy gave evidence of silymarin and phospholipid interaction, which can help in enhancing permeation and hence the bioavailability of silymarin. In vivo studies using acute carbon tetrachloride administration produced a significant increase in serum SGPT level that was significantly reduced by the liposomal formulation and the silymarin suspension. The liposomal formulation significantly ( P < 0.001) reduced the SGPT level compared to suspension. Histopathological examination revealed similar effects with silymarin liposomes and suspension in the improvement of the necrosed area induced by carbon tetrachloride.
Wu et al . prepared a lipid-based SMEDDS, using ethyl linoleate (oil phase), tween 80 (surfactant), and ethanol (cosurfactant).  In vitro drug release studies carried out using the dialysis bag method showed the release of silymarin from SMEDDS as incomplete and having sustained characteristics. The relative bioavailability of SMEDDS was enhanced by an average of 1.88-fold and 48.82-fold from silymarin PEG 400 solution and suspension, respectively. The high bioavailability from the SMEDDS was attributed to its promotion of lymphatic transport  and the presence of long-chain oil, which promotes lipoprotein synthesis and subsequent lymphatic absorption.
The excellent hepatoprotective activity of silymarin, besides its immunomodulatory, antioxidant, and anti-inflammatory activities, as evident by a number of studies cited above, makes it a very promising drug of natural origin. Its good safety profile, easy availability, and low cost are added advantages. It may prove superior to polyherbal formulations in the near future because of its better standardization, quality control, and freedom from microbial and metal contamination.
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