Investigation of protective effects of gypsophila bicolor extracts agai̇nst paracetamole-induced liver damage in rats

Drug-induced liver damage is one of the most common causes of liver damage and is a global health problem that causes hundreds of thousands of deaths around the world. However, today, a satisfactory mechanism or solutions to prevent drug-induced liver cell damage have not yet been found. In this context, although it is known that the Gypsophila genus, of which Gypsophila bicolor is a member, is used as herbal medicine in world folk medicine, there is no FDA-approved saponin-based drug. For this purpose, in recent years, nanoparticulate drug delivery systems are being investigated and new solutions are being investigated by using nanotechnology in order to increase the effectiveness of biopharmaceuticals, which have significant side effects, at the cellular level, to reduce their undesirable side effects, to increase their bioavailability and therefore their therapeutic effectiveness, and to provide effective and safe treatment is being developed. Transferring these systems to the healthcare field appears to be a promising light in the development of new diagnosis and treatment methods. In light of this information in this study, it was aimed to biochemically, histopathologically and molecularly determine the hepatoprotective effect of Gypsophila bicolor plant extract and nanoparticle-encapsulated nanoparticle forms against liver damage on paracetamol-induced hepatotoxicity in experimental animals. By formulating this plant extract in the form of solid lipid nanoparticles (SLN), it is aimed to create stable, highly effective systems with low side effects. In the first stage of the study, solid lipid nanoparticle (GBE-SLN) loaded with Gypsophila bicolor extract was prepared by microemulsion and ultrasonication method. According to the laser diffraction analysis, these particles are distributed in nano-size and have a negative charge, according to FT-IR (infrared spectroscopy) analysis, Gypsophila bicolor extract is encapsulated in solid lipid nanoparticles, according to SEM images, it is spherical and nano-sized, and according to TGA analysis, it is GBE-SLN. It was concluded that it is stable against temperature. In the second stage of the study, in vivo studies were carried out on rats with paracetamol-induced liver damage. Before experimental application, appropriate doses for plant extracts were determined by acute toxicity testing. 64 male Wistar albino rats used in the 21-day treatment were randomly divided into 8 groups (n = 8). All groups except the normal control group received orally administered Paracetamol solution, prepared in 2 ml of 1X PBS (containing 1% CMC), at a dose of 2 g/kg, administered via gavage, 1 hour after the final application. Applications in other groups, except the control, were also carried out orally. Serum and tissue samples of the rats were obtained 24 hours after paracetamol application. Liver function tests in serum samples and antioxidant enzyme activities (CAT, SOD, GSH-Px, GR and GST) and GSH and MDA levels in liver tissue were measured. In addition, the expression level of TNF-α in liver tissues and histological changes of liver tissues were also examined. iv When examining the obtained results, it was observed that the amounts of ALT, AST and ALP, which increased with paracetamol toxicity, decreased significantly in the groups where Gypsophila bicolor extract and nanoparticle forms were applied. Again, according to the biochemical analysis, we observed that enzyme activities (CAT, SOD, GSH-Px, GR and GST) and GSH levels decreased MDA levels increased in the group receiving paracetamol, whereas these enzyme activities and GSH levels increased and MDA levels decreased in the groups given Gypsophila bicolor plant extract and nanoforms. In the histopathological examination performed on rat liver tissues to which solid lipid nanoparticle loaded with Gypsophila bicolor extract was applied at a dose of 100 mg/kg, almost no pathological findings were found and the most similar results to the control group were obtained. We observed that paracetamol significantly increased the expression of TNF-α in the liver tissue of rats compared to the control group, revealing the inflammatory reaction involved in paracetamol-induced liver damage, whereas Gypsophila bicolor extract significantly inhibited the overexpression of TNF-α in the groups (GBN50 ve GBN100) administered in the form of solid lipid nanoparticles. In conclusion; we can say that the application of solid lipid nanoparticles loaded with Gypsophila bicolor extract alleviates liver damage by reducing the expression level of TNF-α, one of the pro-inflammatory cytokines, against paracetamol-induced liver damage, significantly improving liver morphology and activating the antioxidant system, and therefore may have a hepatoprotective and antioxidant effect. It may provide a basis for future research, especially on the treatment of drug-induced liver damage.