Sadauskienė, Ilona

Išeminė širdies liga (IŠL) išlieka viena svarbiausių širdies ir kraujagyslių ligų, siejamų su didele sergamumo ir mirtingumo našta. Jos vystymąsi lemia tarpusavyje glaudžiai susiję kraujagyslių endotelio funkcijos sutrikimai, uždegimas, oksidacinis stresas, trombozė ir renino-angiotenzino sistemos (RAS) pusiausvyros pokyčiai. Greta klasikinių RAS mechanizmų vis didesnę reikšmę įgyja CYP4F2–20-HETE–GPR75 signalinis kelias, tačiau jo vaidmuo skirtingų audinių ląstelėse ir klinikinėje IŠL eigoje nėra pakankamai apibrėžtas. Šio darbo tikslas buvo ištirti RAS inhibitorių, žemo dažnio ultragarso ir CYP4F2–20-HETE–GPR75 ašies moduliacijos poveikį endotelio, inkstų ir kepenų ląstelių funkcijai bei įvertinti su RAS–ACE2 ir eikozanoidų metabolizmu susijusių biožymenų klinikinę reikšmę sergant skirtingomis IŠL formomis. Tyrimo metu kompleksiškai įvertinta terapinės RAS moduliacijos, hemostazę veikiančių vaistų ir žemo dažnio ultragarso sąveika su CYP4F2–20-HETE–GPR75 ašimi, sujungiant kelių tipų ląstelių modelių ir pacientų klinikinius duomenis. Nustatyta, kad RAS ir hemostazę veikiantys vaistai keičia CYP4F2 raišką, 20-HETE aktyvumą, uždegimo žymenų raišką ir funkcinius ląstelių atsako rodiklius. Žemo dažnio ultragarsas sustiprino dalį RAS inhibitorių sukeliamų palankių pokyčių endotelio ir inkstų ląstelėse, įskaitant ląstelių migraciją ir azoto oksido susidarymą. 20-HETE mažino endotelio, inkstų ir kepenų ląstelių metabolinį aktyvumą bei didino GPR75 receptoriaus raišką, patvirtindama šios signalinės ašies reikšmę ląstelių funkcijos reguliacijai. Klinikinėje tyrimo dalyje nustatyta, kad ACE2 ir 20-HETE koncentracijos skiriasi tarp stabilios krūtinės anginos, ūminio koronarinio sindromo ir palyginamosios grupės, o mažiausios jų reikšmės būdingos ūminiu koronariniu sindromu sergantiems pacientams. 20-HETE pasižymėjo didesne diagnostine verte diferencijuojant stabilią ir ūminę IŠL formas. Gauti rezultatai papildo žinias apie RAS ir CYP4F2–20-HETE–GPR75 ašių sąveiką bei pagrindžia tolesnius 20-HETE, kaip perspektyvaus IŠL biožymens, tyrimus.

Background: Oxidative stress is a condition in which the balance between the formation of free radicals and the organism’s antioxidant defence system is disrupted. Free radicals are formed in the body during natural processes such as digestion and respiration. Production can also be increased by environmental factors such as ultraviolet radiation, toxins, and heavy metal ions. Excessive amounts of free radicals cause damage to cell membranes, disrupt cell reproduction, block energy production, damage DNA, and cause its modifications. Oxidative stress is associated with the development of many chronic degenerative diseases. Aim(s): analyse the changes of leukoformula in blood smear during oxidative stress using blood samples from laboratory mice. Methods: 16 male BALB/c mice were studied. The mice were divided into two groups of 8. In each group, they were color-coded at different body sites for their differentiation. The first group of mice was called the control group, in which all mice were given physiological solution orally every day for a month. The second group was called the oxidative stress group, in which mice were given aluminium chloride (AlCl3) every day, thus causing oxidative stress in the mice. AlCl3 is a compound that increases the formation of free radicals and damages the antioxidative system. It is widely used in experimental models of oxidative stress. Blood smears were obtained and divided into groups. They were examined under a microscope with a 100x magnification objective with immersion oil. Cells were counted from the edge in a row until 300 cells were counted. Statistical data analysis was performed with the SPSS program, using oneway ANOVA, and checked with the LSD criterion. Data was presented graphically using Microsoft Excel. Data were considered statistically significant if p<0.05. Results: The number of segmented neutrophils in the oxidative stress group is 30.04%, and in the control group, 9.75% less (p<0.001). This shows an activated inflammatory response and a possible immune system reaction to oxidative stress. The number of monocytes in the control group is 6%, and in the oxidative stress group, 1.27 times less (p<0.05). Decreased monocyte numbers may indicate their migration to damaged tissues. The number of large lymphocytes in the control group is 2.34 times higher than in the oxidative stress group (p<0.01). The number of small lymphocytes in the control group is 7.79% more than in the oxidative stress group (p<0.01). A decrease in lymphocytes may be related to immune system suppression or their increased use during oxidative stress. Conclusion: Oxidative stress causes significant changes in leukoformula associated with inflammatory and immune system mechanisms. These changes are important in explaining the role of oxidative stress in the disease’s pathogenesis and assessing its impact on the immune system.

Background: Bone marrow is the primary hematopoietic organ in mammals and remains highly sensitive to oxidative imbalance. Excessive production of reactive oxygen species (ROS) disrupts hematopoietic homeostasis and promotes inflammatory alterations within the marrow microenvironment. Natural compounds with antioxidant properties may exert protective or modulatory effects on haematopoiesis. Glycyrrhiza glabra (commonly known as liquorice), a medicinal plant widely recognised for its antioxidant and anti-inflammatory activity, has been suggested to influence immune and hematopoietic functions. Aim(s): To evaluate and compare bone marrow cytomorphological changes in BALB/c mice under oxidative stress conditions and following administration of Glycyrrhiza glabra extract. Methods: Male BALB/c mice were randomly divided into groups: (1) control (physiological saline), (2) oxidative stress (AlCl₃ solution), and (3) Glycyrrhiza glabra extract (daily oral herbal extract administration). Mice were weighed every morning. Oxidative stress manifestations appeared within 30 minutes after AlCl₃ administration. Bone marrow smears were obtained from sternums. For each sample, 500 nucleated cells were counted and classified into myeloid and erythroid lineages. Megakaryocytes were evaluated in 10 high-power fields (HPF). The myeloid-to-erythroid (M:E) ratio was calculated. Results: Control animals exhibited balanced haematopoiesis, with segmented neutrophils predominating and active erythropoiesis represented by rubricytes and metarubricytes. The mean M:E ratio was 1.69, corresponding to physiological reference values (1.6–1.9). In the oxidative stress group, a marked increase in segmented neutrophils and their precursors (promyelocytes, metamyelocytes, myeloblasts) indicated stimulated granulopoiesis. Megakaryocyte numbers reached 15/10 HPF, representing a 1.9-fold rise compared to controls. The M:E ratio increased to 2.96, reflecting hematopoietic imbalance and an inflammatory marrow response. In the liquorice group, bone marrow cytology largely resembled the control pattern. Segmented neutrophils predominated, consistent with the BALB/c strain. A slightly elevated eosinophil count compared to controls may suggest a mild hypersensitivity response to the extract. Lymphocyte numbers were moderately reduced. Both erythropoiesis and granulopoiesis remained balanced, with an M:E ratio of 1.63, within physiological limits. Conclusion: AlCl₃-induced oxidative stress significantly alters bone marrow cytomorphology, promoting granulopoiesis and increasing the M:E ratio. In contrast, Glycyrrhiza glabra administration maintains overall hematopoietic balance, with minor immunomodulatory effects. These findings confirm the sensitivity of bone marrow to oxidative stress and suggest a potential modulatory role of antioxidant herbal compounds in maintaining hematopoietic homeostasis.

Introduction. Exposure to environmental and lifestyle stressors (e.g., smoking, air pollution) can enhance oxidative stress, resulting in a higher need for antioxidants. Licorice (Glycyrrhiza glabra L.) roots are a potent source of natural antioxidants, primarily triterpenoid saponins and flavonoids, that exhibit antioxidant and anti-inflammatory effects. However, their biological efficacy is limited by poor stability and low membrane permeability. Liposome-based encapsulation may increase the bioavailability of these herbal constituents. Aim. This study aims to evaluate the effect of liposomes containing licorice (Glycyrrhiza glabra L.) root extract on reduced glutathione and malondialdehyde concentrations in the blood of mice. Materials and methods. The experiments were conducted on 4–6-week-old BALB/c laboratory mice divided into five groups (n=8): 1) control, 2) treated with aluminum solution 3) treated with liposomes containing Glycyrrhiza glabra L. root extract, 4) treated with aluminum solution and liposomes containing Glycyrrhiza glabra L. root extract, 5) treated with phospholipid suspension. Liposomes and phospholipid suspension (10 mL/kg) were administered orally via gavage. An aluminum solution (7.5 mg Al³⁺/kg) was delivered intraperitoneally to induce oxidative stress. Spectrophotometric analysis was used to determine reduced glutathione and malondialdehyde levels in the blood of mice after 21 days of daily exposure to the substances. Statistical significance was determined using one-way analysis of variance (ANOVA) and the unpaired Student’s t-test. The level of significance was taken as a value of p≤0.05. The data were analyzed using Microsoft Excel 2016 (Microsoft Corporation, Redmond, WA, USA). Results and discussion. Exposure to aluminum led to a statistically significant (p≤0,05) decrease in blood GSH concentrations and a significant (p≤0.05) increase in MDA concentrations compared to the control group. The results showed that liposomes containing Glycyrrhiza glabra L. root extract following aluminum exposure significantly (p≤0.05) increased reduced glutathione levels and significantly decreased (p≤0.05) malondialdehyde levels in the blood of mice compared to the aluminum-exposed group. Meanwhile, administration of liposomes containing Glycyrrhiza glabra L. root extract under normal physiological conditions had no effect (p>0,05) on the concentrations of reduced glutathione and malondialdehyde concentrations in the blood of mice. Conclusions. Treatment with Glycyrrhiza glabra L. root extract-loaded liposomes enhanced blood reduced glutathione and malondialdehyde levels under oxidative stress by aluminum, demonstrating their antioxidative activity.

Background: Various stressors (smoking, air pollution, etc.) can induce oxidative overload, increasing the body’s demand for antioxidants. Glycyrrhiza glabra L. roots are rich in triterpenoid saponins and flavonoids with antioxidant and anti-inflammatory activities (1). However, their oral bioavailability is limited by poor stability and low membrane permeability. Liposomal encapsulation may enhance the bioavailability of these compounds. Aim: The aim of this study was to assess the effect of liposomes containing Glycyrrhiza glabra L. root extract on brain glutathione (GSH) and malondialdehyde (MDA) concentrations in mice. Methods: Mice were divided into five groups (n=8): 1) control, 2) exposed to aluminum solution 3) exposed to liposomes with Glycyrrhiza glabra L. root extract, 4) exposed to aluminum solution and liposomes with Glycyrrhiza glabra L. root extract, 5) exposed to phospholipid suspension. Liposomes and phospholipid suspension (10 mL/kg) were administered by oral gavage, while an aluminum solution (7.5 mg Al³⁺/kg) was injected intraperitoneally to induce oxidative stress. Brain GSH and MDA levels in mice were determined spectrophotometrically after 21 days of daily exposure. Results: Aluminum exposure resulted in a statistically significant decrease in brain GSH concentrations and a statistically significant increase in MDA concentrations compared to the control group. In contrast, liposomes containing Glycyrrhiza glabra L. root extract significantly reversed the effects of aluminum by increasing GSH levels and reducing MDA levels compared with the aluminumexposed group, restoring both parameters to control values. Conclusion: Glycyrrhiza glabra L. root extract–loaded liposomes increased brain GSH levels and decreased MDA levels in mice under oxidative stress, indicating their antioxidative activity.

Įžanga. Oksidacinį stresą sukeliantys veiksniai (oro tarša, rūkymas ir kt.) lemia didesnį antioksidantų poreikį organizme. Glycyrrhiza glabra L. šaknyse daugiausia aptinkama triterpeninių saponinų ir flavonoidų, natūralių antioksidantų, kuriems būdingas ne tik antioksidacinis aktyvumas, bet ir kiti biologiniai veikimo būdai, tokie kaip uždegimą mažinantis, gastroprotekcinis, antimikrobinis poveikis [1]. Įkapsuliuojant į liposomas šias išorės veiksniams jautrias medžiagas galima pagerinti junginių bioprieinamumą bei pasiekti tikslesnį jų nukreipimą į atitinkamas organizmo vietas [2]. Tikslas. Įvertinti liposomų su paprastųjų saldymedžių (Glycyrrhiza glabra L.) šaknų ekstraktu poveikį redukuoto glutationo ir malondialdehido koncentracijoms pelių kepenyse. Metodai. Tyrimas atliktas su 4-6 savaičių amžiaus BALB/c linijos pelėmis (n=40). Pelės suskirstytos į 5 grupes pagal instiliuojamas medžiagas: 1) kontrolinę, 2) veikiamą aliuminio tirpalu, 3) veikiamą liposomų su Glycyrrhiza glabra L. šaknų ekstraktu, 4) veikiamą aliuminio tirpalu ir liposomų su Glycyrrhiza glabra L. šaknų ekstraktu, 5) veikiamą fosfolipidų suspensija. Oksidacinio streso sukėlimui naudotas AlCl3 tirpalas (7,5 mg/kg). Po 21 dieną trukusio tyrimo su pelėmis, nustatytos redukuoto glutationo (GSH) ir malondialdehido (MDA) koncentracijos pelių kepenyse taikant spektrofotometrijos metodą. Rezultatai. AlCl3 tirpalas statistiškai reikšmingai padidino MDA koncentraciją (p ≤ 0,05) ir statistiškai reikšmingai sumažino (p ≤ 0,05) GSH koncentraciją pelių kepenyse, lyginant su kontroline grupe. Statistiškai reikšmingas MDA koncentracijos sumažėjimas, lyginant su aliuminio tirpalu paveiktų pelių grupe, nustatytas pelių grupėje, kuriai buvo instiliuojamos liposomos su Glycyrrhiza glabra L. šaknų ekstraktu (p ≤ 0,05). Tyrimo metu taip pat nustatyta, jog liposomos su Glycyrrhiza glabra L. šaknų ekstraktu statistiškai reikšmingai sumažina oksidacinio streso metu padidėjusį MDA kiekį kepenyse (p ≤ 0,05), tačiau statistiškai reikšmingos įtakos MDA koncentracijai, lyginant su kontroline grupe, neturi (p > 0,05). Liposomos su Glycyrrhiza glabra L. šaknų ekstraktu statistiškai reikšmingai padidino GSH koncentraciją (p ≤ 0,05) pelių kepenyse, lyginant tiek su kontroline grupe, tiek su aliuminio tirpalu paveiktų pelių grupe. Ta pati tendencija pastebėta pelių grupėje, kuriai buvo girdomos liposomos su Glycyrrhiza glabra L. šaknų ekstraktu prieš tai sukėlus oksidacinį stresą aliuminio tirpalu. Išvados. Tyrimo metu nustatyta, jog liposomos su Glycyrrhiza glabra L. šaknų ekstraktu geba mažinti lipidų peroksidacijos žymens MDA koncentraciją ir padidinti GSH koncentraciją pelių kepenyse esant oksidaciniam stresui, o tai rodo galimą šių liposomų hepatoprotekcinį poveikį.

Glioma is the most common tumor of the central nervous system, originating from glial cells or their precursors, and accounts for approximately 80% of all malignant brain tumors. Astrocytoma develops as a result of mutations in star-shaped astrocyte cells and constitutes about 75% of all gliomas, among which grade IV astrocytoma and glioblastoma are the most frequently diagnosed. The prognosis for patients diagnosed with this grade of brain tumor is extremely poor, with an overall 5-year survival rate of only 5%. Catalase (CAT) is an enzyme predominantly localized in peroxisomes and plays a crucial role in protecting cells against oxidative stress by preventing the accumulation of hydrogen peroxide (H₂O₂). Elevated CAT expression levels have been reported in various cancer tissues compared to their normal counterparts. In gliomas, catalase appears to be constitutively overexpressed relative to astrocytes. However, the molecular mechanisms regulating CAT expression in astrocytoma remain not fully elucidated. Metal homeostasis is essential for the proper functioning of the brain, which is particularly vulnerable to toxic environmental pollutants. The balance of metals within the brain is maintained through the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier. Based on the ability of metals to cross the BBB, it has been hypothesized that prolonged exposure to certain metals may increase the risk of brain cancer. Nevertheless, no clear association between metal exposure and brain cancer development has been established. The aim of the study was to evaluate the correlations between catalase enzymatic activity and the concentrations of selected trace elements in the blood of patients diagnosed with astrocytoma. The study material consisted of blood and plasma samples obtained from patients diagnosed with astrocytoma. Catalase activity was determined in erythrocytes spectrophotometrically. Copper and zinc concentrations were measured in blood plasma, while selenium concentration was determined in whole blood. Trace element concentrations were determined using inductively coupled plasma mass spectrometry (ICP-MS). The experiments showed that catalase activity increased significantly - by 12% seven days after surgical removal of the tumor.

Background: Bone marrow (medulla ossium) is the principal hematopoietic organ in mammals, accounting for 1.5–2% of body weight in mice. Oxidative stress, defined as an imbalance between the production of reactive oxygen species (ROS) and antioxidant defense, is not only a physiological byproduct of metabolism but also a key pathological factor when excessive. Elevated ROS can disrupt bone marrow homeostasis, impair hematopoietic balance, and promote inflammatory responses. Aim: To investigate in vivo the cytomorphological alterations of bone marrow under oxidative stress in BALB/c mice. Methods: Male BALB/c mice were randomly allocated into two groups: (1) control (physiological saline) and (2) oxidative stress (AlCl₃ solution). Oxidative stress symptoms appeared within 30 minutes of AlCl₃ administration. Bone marrow smears were prepared from femurs. For each specimen, 500 cells were counted and classified into myeloid and erythroid lineages. Megakaryocytes were quantified across 10 high-power fields (HPF). The myeloid-to-erythroid (M:E) ratio was calculated for comparative evaluation. Results: Control animals showed a physiologically balanced hematopoietic profile, with segmented neutrophils predominating and active erythropoiesis represented by rubricytes and metarubricytes. The mean M:E ratio was 1.69, consistent with reported physiological values (1.6–1.9). In the oxidative stress group, a significant increase in segmented neutrophils and their precursors (promyelocytes, metamyelocytes, myeloblasts) indicated enhanced granulopoiesis. Megakaryocytes averaged 15/10 HPF, a 1.9-fold increase compared to controls. The M:E ratio rose to 2.96, demonstrating hematopoietic imbalance and an inflammatory marrow environment. Conclusion: AlCl₃-induced oxidative stress significantly alters bone marrow cytomorphology in BALB/c mice, characterized by increased granulopoiesis, elevated megakaryocyte density, and a pathologically high M:E ratio. These findings confirm that oxidative stress profoundly disrupts hematopoietic homeostasis and highlight the suitability of this in vivo model for studying ROS-mediated hematological alterations.

Selenium is an essential microelement involved in antioxidant defence, redox regulation, and cellular signalling. Both its inorganic form, sodium selenite (Na₂SeO₃), and organic form, selenomethionine (SeMet), influence gene expression related to oxidative stress, apoptosis, and cell survival. The brain and liver, being highly vulnerable to oxidative stress, are critical tissues in understanding selenium’s biological effects. This study evaluates the impact of sodium selenite and selenomethionine on gene expression in these tissues, focusing on genes involved in redox regulation (MsrB1), apoptosis (Caspase-3), and cell cycle control (GADD45). The study was performed on 4-6-week-old BALB/c mice, which were divided into 4 experimental groups, that were given solutions of different concentrations of SeMet or Na₂SeO₃ (0.2 and 0.4 mg Se/kg body weight) for 8 weeks, and a control group that had free access to tap water. Then, mRNA was extracted from mouse liver and brain tissues and used to synthesize cDNA for RT-PCR analysis. The results show that MsrB1 expression in both brain and liver was higher in mice receiving 0.2 mg Se/kg of either SeMet or Na₂SeO₃ compared to the control group and those receiving 0.4 mg Se/kg. Caspase-3 expression in brain and liver was increased in mice receiving 0.2 mg Se/kg of Na₂SeO₃, but lower in those receiving 0.2 mg Se/kg of SeMet compared to the control group and those given 0.4 mg Se/kg of either selenium form. GADD45 expression in the brain and liver was elevated in mice given 0.2 mg Se/kg of Na₂SeO₃, while in SeMet-treated mice, GADD45 expression in the liver increased with higher concentrations, whereas in the brain, it decreased with increasing SeMet dosage. Overall, Na₂SeO₃ may promote stronger oxidative stress defence and apoptosis regulation, while SeMet could be more selective in its influence on apoptosis and cell cycle control, depending on tissue type.

Introduction. Glycyrrhiza glabra L., is a member of the Fabaceae family and one of the most valuable plants in the world for commerce. The chemical composition of G. glabra is glycyrrhizin, glycyrrhetic acid, isoliquiritin, isoflavones, etc., and their derivatives have been reported for several pharmacological activities like antiulcer, anticancer, anti-inflammatory, antidiabetic and antioxidant [1]. Materials and methods. The study was performed on 6-week-old BALB/c mice (n=32). The AlCl3 solution was used to induce oxidative stress. Catalase activity (CAT) in the brain and liver of mice were determined spectrophotometrically after 21 days of administration of Glycyrrhiza glabra L. extract. Results and their discussion. The results showed that aluminium statistically significant increased CAT activity in mice liver by 23.34% and in brain by 68.69%, compared to control group (36.25 ± 2.37 U/mg protein and 65.30 ± 5,95 U/mg protein respectively, p ≤ 0,05). Glycyrrhiza glabra L. extract after aluminium exposure increased CAT activity in liver by 10.68% and in brain by 15.32% compared to control group, but these results were not statistically significant (32.53 ± 1.81 U/mg protein and 44.64 ± 3.84 U/mg protein, p > 0,05). Compared to aluminium-treated group, Glycyrrhiza glabra L. extract statistically insignificant (p > 0,05) reduced CAT activity in both the liver by 10.26% and in brain by 31.64% after aluminium exposure. These results showed that the administration of Glycyrrhiza glabra L. extract did not show any statistically significant decrease in CAT activity in the liver and brain after aluminium exposure.