Fermented formulation of Silybum marianum seeds: Optimization, heavy metal analysis, and hepatoprotective assessment.

BACKGROUND: Fermented formulations are extensively used in Ayurveda due to several benefits like improved palatability, bioavailability, pharmacological potential, and shelf life. These formulations can also quench the heavy metals from the plant material and thus reduce the toxicity. Seeds of Silybum marianum (L.) Gaertn. are widely used for the management of many liver diseases. STUDY DESIGN AND METHODS: In the present study, we developed a novel fermented formulation of S. marianum seeds and evaluated parameters like safety (heavy metal analysis) and effectiveness (hepatoprotective). As the developed formulation's validation is crucial, the critical process variables (time, pH, and sugar concentration) are optimized for alcohol and silybin content using the Box-Behnken design (BBD). RESULTS: The response surface methodology coupled with BBD predicted the optimized conditions (fermentation time (28 days), pH 5.6, and sugar concentration (22.04%)) for the development of a fermented formulation of the selected herb. Moreover, the alcohol content (6.5 ± 0.9%) and silybin concentration (26.1 ± 2.1%) were confirmed in optimized formulation by GC-MS and HPTLC analysis. The optimized formulation was also analyzed for heavy metals (Pb, As, Hg, and Cd); their concentration is significantly less than the decoction of herbs. Further, the comparative evaluation of the developed formulation with the marketed formulation also confirmed that the fermented formulation's silybin concentration and percentage release were significantly enhanced. In addition, the developed fermented formulation's percentage recovery of HepG2 cell lines after treatment with CCl4 was significantly improved compared with the marketed formulation. CONCLUSION: It can be summarized that the developed fermented formulation improves safety and effectiveness compared to other market formulations. Finally, it can be concluded that the developed fermented formulation could be further explored as a better alternative for developing Silybum marianum preparation.

Rosuvastatin and retinoic acid may act as 'pleiotropic agents' against ß-adrenergic agonist-induced acute myocardial injury through modulation of multiple signalling pathways.

Cardiovascular disorders constitute the principal cause of deaths worldwide and will continue as the major disease-burden by the year 2060. A significant proportion of heart failures occur because of use and misuse of drugs and most of the investigational agents fail to achieve any clinical relevance. Here, we investigated rosuvastatin and retinoic acid for their "pharmacological pleiotropy" against high dose ß-adrenergic agonist (isoproterenol)-induced acute myocardial insult. Rats were pretreated with rosuvastatin and/or retinoic acid for seven days and the myocardial injury was induced by administering isoproterenol on the seventh and eighth day. After induction, rats were anaesthetized for electrocardiography, then sacrificed and different samples were collected/stored for various downstream assays. Myocardial injury with isoproterenol resulted in increased cardiac mass, decreased R-wave amplitude, increased QRS and QT durations; elevated levels of cardiac markers like cTnI, CK-MB, ALT and AST; increased lipid peroxidation, protein carbonylation and tissue nitric oxide levels; decreased endogenous antioxidants like SOD, CAT, GR, GST, GPx and total antioxidant activity; increased inflammatory markers like TNF-α and IL-6; decreased the mRNA expression of Nrf2 and Bcl-2; increased the mRNA expression of Bax, eNOS and iNOS genes. Pretreatment with rosuvastatin and/or retinoic acid mitigated many of the above biochemical and pathological alterations. Our results demonstrate that rosuvastatin and retinoic acid exert cardioprotective effects and may act as potential agents in the prevention of ß-adrenergic agonist-induced acute myocardial injury in rats. Cardioprotective potential of rosuvastatin and retinoic acid could be attributed to their influence on the redox pathways, immunomodulation, membrane stability, Nrf2 preservation, iNOS and Bax expression levels. Thus, they may act directly or indirectly at various steps, the breakpoints, in the pathophysiological cascade responsible for cardiac injury. Our study gives insights about the pharmacological pleiotropism of rosuvastatin and retinoic acid.

The toxicity of four native Indian plants: effect on AChE and acid/alkaline phosphatase level in fish Channa marulius.

The latex of four plants viz. Euphorbia royleana, Jatropha gossypifolia (Euphorbiaceae), Nerium indicum and Thevetia peruviana (Apocynaceae) caused significant reduction in acid/alkaline phosphatase activity and anti-acetylcholinesterase activity in nervous tissue of freshwater air breathing fish Channa marulius. The reduction in the activity of both phosphatases and AChE were time as well as dose dependent.

Toxic effect of two common Euphorbiales latices on the freshwater snail Lymnaea acuminata.

The aqueous and serially purified latex extracts of plants Euphorbia pulcherima and Euphorbia hirta (family Euphorbiaceae) have potent molluscicidal activity. Sub-lethal doses (40 and 80% of LC(50)) of aqueous and partially purified latex extracts of both the plants also significantly alter the levels of total protein, total free amino acid, nucleic acid (DNA and RNA) and the activity of enzyme protease and acid and alkaline phosphatase in nervous tissue of the snail Lymnaea acuminata in time and dose dependent manner. E. pulcherima and E. hirta are common medicinal plants of family Euphorbiaceae. E. pulcherima is useful for a variety of conditions, such as rheumatism, snakebite, asthma, obstipation, and skin-diseases. While, E. hirta is also used in cough, asthma, colic, dysentery, and genito urinary diseases.

Piscicidal effect of some common plants of India commonly used in freshwater bodies against target animals.

Mortality caused by the aqueous extracts of leaf and stem bark of four plants belonging to family Euphorbiaceae and Apocynaceae against freshwater fish Channa punctatus has been reported. It was found that dilute aqueous solutions of leaf and stem bark were active in killing the fishes. The toxic effect of stem bark of all the plants were time as well as dose dependent. There was significant negative correlation between LC50 and exposure periods. Thus, the LC50 values of stem bark extracts of Euphorbia royleana, Jatropha gossypifolia, Nerium indicum and Thevelia peruviana were decreased from 0.050 g/l (24 h) > to 0.020 g/l (96 h); 4.61 g/l (24 h) > to 4.34 g/l (96 h); 0.097 g/l (24 h) > to 0.041 g/l (96 h) and 4.05 g/l (24 h) > to 3.17 g/l (96 h), respectively. It has been suggested that these plant products cannot be used directly in freshwater bodies, without their detailed studies on long-term effects on non-target organism as well their structure activity relationship.