Agnieszka Zakrzeska

Central nervous system disorders, such as Alzheimer’s disease, Parkinson’s disease, and multiple sclerosis, are associated with complex pathophysiological mechanisms involving oxidative stress, inflammation, and protein misfolding. According to the literature, betulin, a natural compound derived from the bark of birch trees, demonstrates promising neuroprotective effects. This study investigates the neuroprotective potential of betulin and its complex with cyclodextrin, referred to as Betula Forte, using an in vitro model of differentiated SH-SY5Y neuroblastoma cells. Specifically, the study explores the antioxidant and antiapoptotic properties of these compounds under oxidative stress induced by hydrogen peroxide (H₂O₂). Our results indicated that Betula Forte exhibited lower cytotoxicity compared to betulin alone. Both substances enhanced cell viability in pre-incubation and co-incubation models, with Betula Forte showing superior efficacy under severe oxidative stress. Additionally, both substances exhibited protective effects against H₂O₂-induced oxidative stress and apoptosis, as evidenced by reduced levels of reactive oxygen species (ROS) and a lower number of apoptotic cells compared with the H₂O₂-treated cells. These findings suggest that Betula Forte, due to lower cytotoxicity, may offer a more effective neuroprotective strategy than betulin alone, highlighting its potential as a therapeutic agent for neurodegenerative diseases.

The multifunctional characteristics of Phallus impudicus extract encourage to conduct research for its potential use in medical applications. Diabetes mellitus is a metabolic disease, accompanied by chronic hyperglycemia caused by absolute or relative insulin deficiency. To purpose of this study was to show the potential role of Phallus impudicus extract in accelerating of wound healing in diabetes rats. For the induction of diabetes, streptozotocin (STZ, concentration 100 mg/ml) at a dose of 30 mg/kg b.w. in citrate buffer, pH 4.5 was used. Wounds were excised by cutting a 10-mm-diameter flap of full-thickness skin with a barb under inhalation anaesthesia, 5% isoflurane under aseptic conditions. Wounds were treating with standard used preparation and with prototype product containing an alcoholic extract of Phallus impudicus. A number of metabolites which promote wound healing including amino acids, sugars, organic acids and other compounds were identified in ethanolic extract by GC-MS. In the MTT test, the alcoholic extract of P. impudicus was non-toxic and stimulated fibroblast proliferation, which was also expressed as an increase in DNA and collagen synthesis. A hydrogel containing an extract of the fungus Phallus impudicus was shown to accelerate the period of wound desquamation in the experimental animals compared to the control groups. There was also a statistically significant (p>0.005) reduction in SOD activity and in TBARS compared to the control group, indicating an intensive healing process in which the oxidative enzyme systems play a significant role.

Betulin is natural pentacyclic triterpenoids lupane-structure, which shows a wide range of biological activity, including possibly hypoglycemic effects. Diabetes mellitus is a metabolic disease, accompanied by chronic hyperglycemia caused by absolute or relative insulin deficiency. The main effective drug in the treatment of type 1 diabetes is insulin injections. In the case of type 2 diabetes, it is an adjuvant therapy. To purpose of this study was to show hypoglycemic action of betulin in a model of experimentally induced diabetes in rats. For the induction of diabetes, alloxan at a dose of 170 mg/kg, i.p. was used. The hypoglycemic effect of betulin was evaluated using doses of 50 and 100 mg/kg/day for 30 days. The activity of glucose metabolism enzymes: liver glycogen phosphorylase, hexokinase and liver and skeletal muscle phosphofructokinase in rats, glucose-6-phosphate dehydrogenase (DG-6-P) of the liver was also evaluated. Betulin in a dose-dependent manner normalized of glycemia, increased the basal insulin concentration in the blood serum, increased the activity of carbohydrate metabolizing enzymes and on the gene expression profile of enzymes involved in carbohydrate metabolism. Betulin also significant improved the morphology of the pancreas. In particular, there was an increase in the area of the islands, their diameter, an increase in the number of β cells in the composition of the islands. We conclude that betulin has hypoglycemic properties, improving glycemia and carbohydrate metabolizing enzymes and pancreatic morphology.

Type 2 diabetes mellitus (T2DM) is a complex chronic metabolic disease characterized by long-term hyperglycemia, which is, in turn, resulted from the impaired insulin signaling caused by a combination of insulin resistance or inadequate insulin production. Prevalence and incidence of T2DM are increasing dramatically across the world, and it is accompanied with severe complications and premature mortality of patients with diabetes. Given the fact that synthetic drugs have disadvantages in view of the side effects, the implementation of naturally occurring compounds for diabetes treatment may be a promising alternative. Betulin is a naturally occurring triterpenoid which has been shown to possess the ability of altering body lipids and exert hypoglycemic and hepatoprotective effects. It is suggested that the application of betulin in T2DM may have a favorable effect to ease the severity of diabetic complications. Thus, the aim of the present study was to assess biological effects of betulin in T2DM conditions. Diabetes-induced rats were administered with two different doses of betulin for 28 consecutive days. It was shown that long-term administration of betulin at the doses of 50 and 100 mg/kg/day to the rats prevented diabetes-associated changes in a body weight of the animals, significantly reversed insulin resistance and abolished the impairment of glucose metabolism. It was accompanied with the dose-dependent normalization of serum lipid contents. Histopathological changes and structural abnormalities in the liver of diabetics were restored by the administration of betulin. Also, betulin was able to restrain systemic inflammation detected in diabetic animals according to the altered levels of serum TNFα. Thus, the results obtained in the current study were found to be in agreement with earlier findings on beneficial effects of betulin in conditions pathogenetically close to T2DM. We hypothesized that the ability of betulin to restrain systemic inflammation and to normalize the lipid metabolism can explain improved insulin resistance and glycemic control and it can provide a possible mechanism for the beneficial antidiabetic effects of betulin.

The aim of this study was to evaluate the therapeutic effects of chlorogenic acid (CGA) in rats with advanced alcoholic steatohepatitis. The rats were fed on a high-fat diet and gavaged with ethanol (4 g/kg) for 8 weeks. The livers of ethanol-treated rats showed steatosis; necrosis and mononuclear infiltration; and significant upregulation of the mRNA expression of the prooxidant (Cyp2e1, iNos), lipogenic (Srebp1, Acc), proinflammatory (Tlr4, Nf-κb, TnfA, Il-1B, and Il-6), and profibrogenic (TgfB, Col1, VegfA) genes. Simultaneously, a downregulation of level of Sod and Nrf2 was observed, which was accompanied by increased serum transaminase, TnfA, and serum and liver triglycerides levels. CGA administration (40 and 80 mg/kg, 8 weeks) to ethanol-fed group reduced the liver expression levels of Cyp2e1 and iNos, whereas it markedly enhanced the expression of Sod, Nrf2, and Ho-1. CGA at both doses downregulated the expressions of lipogenic, proinflammatory, and profibrogenic genes, while the expression of Tlr4 was lowered only after the higher dose of CGA. The higher dose of CGA efficiently prevented the progression of alcohol-induced steatosis and reduced inflammation through regulation of the expression of genes encoding the proteins involved in the Tlr4/Nf-κB signaling pathway and fibrosis. The study revealed hepatoprotective and anti-inflammatory effects of CGA through the regulation of expression of genes encoding Cyp2e1/Nrf2 involved in oxidative stress modulation. These results demonstrate CGA as a therapeutic candidate for the prevention and treatment of alcoholic steatohepatitis.

We investigated the role of aldosterone (ALDO) in the development of arterial thrombosis in streptozotocin-induced diabetic rats. To evaluate the effect of endogenous ALDO, the rats underwent adrenalectomy (ADX). ADX reduced the development of arterial thrombosis. A 1 h infusion of ALDO (30 μg/kg/h) enhanced thrombosis in adrenalectomized rats, while this effect was potentiated in diabetic rats. ALDO shortened bleeding time, increased plasma levels of tissue factor (TF) and plasminogen activator inhibitor, decreased plasma level of nitric oxide (NO) metabolites, and increased oxidative stress. Moreover, 2 h incubation of human umbilical vein endothelial cells (HUVECs) with ALDO (10⁻⁷ M) disrupted hemostatic balance in endothelial cells in normoglycemia (glucose 5.5 mM), and this effect was more pronounced in hyperglycemia (glucose 30 mM). We demonstrated that the acute ALDO infusion enhances arterial thrombosis in rats and hyperglycemia potentiates this prothrombotic effect. The mechanism of ALDO action was partially mediated by mineralocorticoid (MR) and glucocorticoid (GR) receptors and related to impact of the hormone on primary hemostasis, TF-dependent coagulation cascade, fibrinolysis, NO bioavailability, and oxidative stress balance. Our in vitro study confirmed that ALDO induces prothrombotic phenotype in the endothelium, particularly under hyperglycemic conditions.

Sodium alginate is a polymer with unique ability to gel with different cross-linking agents in result of ionic and electrostatic interactions. Chitosan cross-linked alginate provides improvement of swelling and mucoadhesive properties and might be used to design sustained release dosage forms. Therefore, the aim of this research was to develop and evaluate possibility of preparing chitosan cross-linked alginate microparticles containing metformin hydrochloride by the spray-drying method. In addition, influence of cross-linking agent on the properties of microparticles was evaluated. Formulation of microparticles prepared by the spray drying of 2% alginate solution cross-linked by 0.1% chitosan was characterized by good mucoadhesive properties, high drug loading and prolonged metformin hydrochloride release. It was shown that designed microparticles reduced rat glucose blood level, delayed absorption of metformin hydrochloride and provided stable plasma drug concentration. Additionally, histopathological studies of pancreas, liver and kidneys indicated that all prepared microparticles improved degenerative changes in organs of diabetic rats. Moreover, no toxicity effect and no changes in rats behavior after oral administration of chitosan cross-linked alginate microparticles were noted.