Probiotyki jako eksperymentalna metoda zapobiegania otyłości: wpływ liczby podawanych szczepów bakteryjnych i ich żywotności

Introduction: a comparative animal study of the efficacy of intermittent short-course administration of lyophilised single-, three-, and live multistrain probiotic on obesity. Methods: We included 70 rats divided into seven groups (n = 10 in each). Rats of group I were intact. Newborn rats of groups II–VII were injected with monosodium glutamate (MSG) (4 mg/g). Rats of group II (MSG-obesity group) were untreated. The group III-V received lyophilised monoprobiotics B. animalis VKL, B. animalis VKB, and L. casei IMVB-7280, respectively. Group VI received the mix of these three probiotic strains. Group VII was treated with multiprobiotic “Symbiter”, which contains 14 live probiotic strains (Lactobacillus, Bifi­dobacterium, Propionibacterium, Acetobacter genera). Results: Neonatal treatment with MSG caused stunted growth, which is why, despite the lack of weight gain dynamics and absence of significant food consumption rate and body weight changes at day 120, we noted the development of obesity in all MSG-obesity rats and in up to 20–70% after probiotic administration. Supplementation of probiotic composition, with preference to live strains, led to a significantly lower prevalence of obesity, and reduction of VAT weight and serum lipid levels as compared to single-strain probiotic. In our comparative single-strain analysis a trend towards more pronounced hypolipidaemic effect and VAT weight reduction was observed for lyophilised L. casei IMVB-7280 as compared to B. animalis VKL and VKB strains. Conclusions: Multistrain formed mutualistic interactions in mixtures and therefore able to share with different metabolites, affect differ­ent receptors and produced various of biologically active compounds which synergistic overall effect greater than the sum of the single effects.  Wstęp: Badania porównawcze na zwierzętach oceniające skuteczność w zapobieganiu otyłości podawania w krótkotrwałych cyklach liofilizowanych preparatów zawierających jeden, trzy lub więcej żywych szczepów probiotycznych. Materiał i metody: Do badania włączono 70 szczurów podzielonych na 7 grup (n = 10 w każdej grupie). Szczury w grupie I nie zostały poddane żadnej interwencji. Noworodkom szczurów w grupach II–VII wstrzyknięto glutaminian jednosodowy (monosodium glutamate, MSG) (4 mg/g). U szczurów z grupy II (z otyłością indukowaną MSG) niż stosowano żadnego leczenia. Szczury w grupach III–V otrzy­mywały liofilizowane probiotyki, odpowiednio B. animalis VKL, B. animalis VKB i L. casei IMVB-7280. Grupie VI podawano mieszankę tych trzech szczepów. W grupie VII stosowano wieloskładnikowy preparat probiotyczny „Symbiter” zawierający 14 żywych szczepów (Lactobacillus, Bifidobacterium, Propionibacterium, Acetobacter). Wyniki: Podanie szczurzym noworodkom MSG spowodowało zahamowanie wzrostu, jednak mimo wolniejszego przyrostu masy ciała i braku istotnych zmian wielkości spożycia karmy i masy ciała po 120 dniach zaobserwowano rozwój otyłości u wszystkich szczurów z otyłością indukowaną MSG i u 20–70% zwierząt otrzymujących probiotyki. Podawanie kompozycji probiotyków, szczególnie żywych szczepów, prowadzi do istotnego zmniejszenia częstości występowania otyłości, ilości tkanki tłuszczowej trzewnej i stężania lipidów w surowicy w porównaniu ze stosowaniem preparatów jednoszczepowych. W analizie porównawczej preparatów jednoskładniko­wych stwierdzono tendencję w kierunku silniejszego działania hipolipemicznego i większej redukcji tkanki tłuszczowej trzewnej w przypadku liofilizowanego szczepu L. casei IMVB-7280 niż szczepów B. animalis VKL i VKB. Wnioski: W preparatach wieloszczepowych powstają wzajemne interakcje umożliwiające wymianę różnych metabolitów, wpływ na różne receptory i produkcję różnych biologicznie czynnych cząsteczek, których ogólny efekt synergistyczny jest większy niż suma efektów jednostkowych.

Dynamic Properties of Skeletal Muscle Contraction in Rats with Diabetes

The study was conducted on 20 white nonlinear male rats, which were divided into 2 groups of 10 animals each. Rats in the first group were used as control. Rats in the second group were induced type I diabetes by intraperitoneal (i.p.) administration of streptozotocin (65 mg/kg). Diabetes in rats was confirmed by the presence of hyperglycemia. For the establishment of nociceptive pain sensation, mechanical nociceptive test and tail-flick test were conducted in rats. Further animals were anesthetized by i.p. administration of Nembutal (40 mg/kg). The study of dynamic properties of muscle contraction was performed under conditions of the tibia muscle activation by using the modulated stimulation of efferent n. tibialis. Streptozotocin (STZ) was injected in rats; as a result, the blood glucose level was increased by 4.4 times (p ≤ 0.001). Pain sensitivity in diabetic rats was suppressed, indicating the development of peripheral neuropathy. In rats with diabetes, biomechanical parameters of tibia muscle contraction such as the maximum force of contraction, the speed of maximum force of contraction, the retention time of maximum force of contraction and integrated power of muscle contraction (it is calculated on the total area of the received force curves) were violated. This prevents adequate implementation motor neuron pools muscular system, which will have significant consequences in accurate positional movements

Профіль сироваткових цитокінів у щурів з експериментальним виразкоутворенням на тлі лікувального введення пролінвмісної сполуки

The purpose was to study the therapeutic properties of the low molecular weight organic compound sodium 2-(2-hydroxyphenoxy) acetyl)-L-prolinate in the conditions of gastric ulceration in rats caused by stress, ethanol, non-steroidal anti-inflammatory drugs (indomethacin and aspirin). The test substance is administered in a dose of 1 mg/kg three times for three days after the ulcerogenic stimulus. It was established a considerable acceleration of the gastric mucosa healing of rats under the influence of studied compound. Its antiulcer properties was associated with a decrease of the proinflammatory cytokines content in blood and increase of the content of anti-inflammatory cytokines and prostaglandin E2.Досліджено терапевтичні властивості нової сполуки 2-(2-гідроксифенокси)ацетил)-L-пролінату натрію при стресовій, етаноловій, індометациновій та аспіриновій моделях виразкоутворення в слизовій оболонці шлунка щурів. Антивиразкові властивості сполуки були асоційовані зі зменшенням вмісту прозапальних цитокінів у крові та збільшенням вмісту антизапальних цитокінів і простагландину Е2

Порівняльна дія сучасних прокінетиків та нанокристалічного діоксиду церію на моторну функцію травного тракту у щурів різного віку

In this paper was carried out a comparative study of action of the cerucal, sennosides and NCD on motor function of the stomach and colon in rats. The experiments were performed by ballonographic method.It is shown that cerucal reduces motor, tonic and phasic indexes, amplitude, frequency of the contractions of the stomach and colon in rats. Sennoides increased only the frequency of the contractions, whereas other parameters of motor activity it decreases. It was found that the NCD has stimulated the amplitude, frequency, phase and motor indexes of spontaneous contractile activity of the stomach and colon in rats of both age groups. Thus, the NDC showed effective prokineticffects than current prokinetics. Based on the NCD can be created a prokineticsa new generation.Встановлено, що нанокристалічний діоксид церію посилював у старих щурів моторну активність шлунка та товстої кишки на відміну від церукалу та сенаде. Нанокристалічний діоксид церію – ефективніший прокі-нетику, ніж сучасні. Отримані результати можуть бути підґрунтям для створення на основі нанокриста-лічногодіоксиду церію протизакрепних засобів нового покоління

Probiotics and smectite absorbent gel formulation reduce liver stiffness, transaminase and cytokine levels in NAFLD associated with type 2 diabetes: a randomized clinical study

Introduction. In double-blind single center randomized clinical trial (RCT), the efficacy of alive probiotics sup­plementation with smectite gel vs. placebo in type 2 diabetes patient with non-alcoholic fatty liver disease (NAFLD) detected on ultrasonography (US) were studied. Material and methods. A total of 50 patients met the criteria for inclusion. They were randomly assigned to receive Symbiter Forte combination of probiotic biomass with smectite gel (250 mg) or placebo for 8-weeks. The primary main outcomes were the change in fatty liver index (FLI) and liver stiffness (LS) meas­ured by shear wave elastography (SWE). Secondary outcomes were the changes in transaminases activity, serum lipids and cytokines levels. Results. All subjects completed the study and received more than 90% of prescribed sachets. In respect to our primary endpoints, FLI and LS insignificant de­crease in both interventional and placebo groups. However, when we compare mean changes across groups from baseline, expressed in absolute values, the reduction of both LS (–0.254 ± 0.85 vs. 0.262 ± 0.77; p = 0.031) were observed. Analysis of sec­ondary outcomes showed that co-administration of probiotic with smectite lead to significant reduction of alanine aminotransferase (ALT), aspartate amino­transferase (AST), total cholesterol, IL-1b, and tumor necrosis factor (TNF-a) after 8 weeks. Conclusion. In this RCT, we confirmed previously re­ported animal data, showing that co-administration of probiotic with smectite manifested with reduction of LS, liver transaminases and chronic systemic inflam­mation

Wpływ Z56822977 na biosyntezę serotoniny w mózgu szczurów z otyłością wywołaną przez podawanie glutaminianu sodu

Wstęp: Badanie przeprowadzono w celu wyjaśnienia wpływu Z56822977 na biosyntezę serotoniny w mózgu szczurów z otyłością wy­wołaną podawaniem glutaminianu sodu (monosodium glutamate, MSG). Materiał i metody: W badaniu wykorzystano 18 samców szczura. Zwierzęta podzielono na trzy grupy: 1 — grupa kontrolna, 2 — grupa MSG, 3 — grupa MSG + Z56822977. Szczurzym oseskom w grupie 2 i 3 podawano podskórnie MSG rozpuszczony w soli fizjologicznej w dawce 4 mg/g masy ciała w objętości 8 μl/g w 2., 4., 6., 8. i 10. dniu życia. Grupie 3 podawano doustnie wodny roztwór Z56822977 w dawce 25 mg/kg w objętości 1 ml/kg. Pierwszą dawkę Z56822977 podawano po ukończeniu 4 tygodni życia, a następnie kontynuowa­no podawanie badanej substancji cyklicznie wedlug schematu tydzień podawania substancji badanej/3 tygodnie przerwy. Zwierzętom z grupy MSG podawano odpowiednio 1 ml/kg wody doustnie. Przez pierwsze 4 miesiące życia szczury otrzymywały standardową karmę. Zmierzono zawartość serotoniny, tryptofanu i 5-hydroksytryptofanu (5-HTr) oraz aktywność hydroksylazy tryptofanowej (tryptophan hydroxylase, TRH), dekarboksylazy aminokwasów (amino acid decarboxylase, AADC) i monoaminooksydazy (MAO) w tkance mózgowej. Wyniki: Wykazano, że podawanie Z56822977 ma pozytywny wpływ na główne wskaźniki otyłości, co odzwierciedlają zmiany podsta­wowych parametrów fizjologicznych i biochemicznych [zmniejszenie masy ciała o 13% vs. MSG (p < 0,05); zmniejszenie wskaźnika masy ciała (body mass index, BMI), wskaźnika Lee oraz masy tkanki tłuszczowej trzewnej odpowiednio o 18%, 7% i 55%, (p < 0,05) w porównaniu z grupą MSG]. Zawartość tryptofanu i serotoniny była istotnie niższa (p < 0,05) u szczurów z otyłością wywołaną przez MSG. W badaniach wykazano, że u otyłych szczurów aktywność MAO zwiększa się o 97% (p < 0,05), a aktywność TRH i AADC odpowiednio o 44% i 53% (p < 0,05). Podawanie Z56822977 powodowało zwiększenie zawartości serotoniny i tryptofanu w mózgach szczurów i przywracało poziom aktywności enzymów (MAO, TRH, AADC) do wartości mierzonych u zwierząt kontrolnych. Wnioski: Wiadomo, że otyłość wiąże się z zaburzeniem syntezy serotoniny w mózgu szczurów. Jednak podawanie Z56822977 prowadzi do normalizacji stężenia serotoniny i tryptofanu oraz przywrócenia prawidłowej aktywności enzymów uczestniczących w biosynte­zie i degradacji serotoniny. Podawanie Z56822977, cząsteczki wpływającej na układ serotoninergiczny, może powodować korzystne efekty w leczeniu otyłości wywołanej przez MSG u szczurów. Można rozważać zastosowanie cząsteczki Z56822977 jako nowego leku stosowanego w otyłości, jednak konieczne są dalsze badania w celu potwierdzenia jej działania

Pharmacological Correction of Stress-Induced Gastric Ulceration by Novel Small-Molecule Agents with Antioxidant Profile

This study was designed to determine novel small-molecule agents influencing the pathogenesis of gastric lesions induced by stress. To achieve this goal, four novel organic compounds containing structural fragments with known antioxidant activity were synthesized, characterized by physicochemical methods, and evaluated in vivo at water immersion restraint conditions. The levels of lipid peroxidation products and activities of antioxidative system enzymes were measured in gastric mucosa and correlated with the observed gastroprotective activity of the active compounds. Prophylactic single-dose 1 mg/kg treatment with (2-hydroxyphenyl)thioacetyl derivatives of L-lysine and L-proline efficiently decreases up to 86% stress-induced stomach ulceration in rats. Discovered small-molecule antiulcer agents modulate activities of gastric mucosa tissue superoxide dismutase, catalase, and xanthine oxidase in concerted directions. Gastroprotective effect of (2-hydroxyphenyl)thioacetyl derivatives of L-lysine and L-proline at least partially depends on the correction of gastric mucosa oxidative balance

Probiotics and smectite absorbent gel formulation reduce liver stiffness, transaminase and cytokine levels in NAFLD associated with type 2 diabetes: a randomized clinical study

Introduction. In double-blind single center randomized clinical trial (RCT), the efficacy of alive probiotics sup­plementation with smectite gel vs. placebo in type 2 diabetes patient with non-alcoholic fatty liver disease (NAFLD) detected on ultrasonography (US) were studied. Material and methods. A total of 50 patients met the criteria for inclusion. They were randomly assigned to receive Symbiter Forte combination of probiotic biomass with smectite gel (250 mg) or placebo for 8-weeks. The primary main outcomes were the change in fatty liver index (FLI) and liver stiffness (LS) meas­ured by shear wave elastography (SWE). Secondary outcomes were the changes in transaminases activity, serum lipids and cytokines levels. Results. All subjects completed the study and received more than 90% of prescribed sachets. In respect to our primary endpoints, FLI and LS insignificant de­crease in both interventional and placebo groups. However, when we compare mean changes across groups from baseline, expressed in absolute values, the reduction of both LS (–0.254 ± 0.85 vs. 0.262 ± 0.77; p = 0.031) were observed. Analysis of sec­ondary outcomes showed that co-administration of probiotic with smectite lead to significant reduction of alanine aminotransferase (ALT), aspartate amino­transferase (AST), total cholesterol, IL-1b, and tumor necrosis factor (TNF-a) after 8 weeks. Conclusion. In this RCT, we confirmed previously re­ported animal data, showing that co-administration of probiotic with smectite manifested with reduction of LS, liver transaminases and chronic systemic inflam­mation

Evaluation of proteolytic activity and serine proteases distribution in plasma from patients with bladder cancer

BackgroundBladder cancer (BC) is an aggressive disease with a poor prognosis. A bladder tumor, like other malignant neoplasms, is characterized by the presence of both cancer cells and stromal cells which secrete cytokines, chemokines, growth factors, and proteolytic enzymes. One such class of proteolytic enzymes are serine proteases, which take part in the tumor microenvironment formation via supporting and contributing to tumor progression. This study aims to evaluate the proteolytic activity and serine protease contribution in plasma from BC patients.MethodsThe research involved patients of Alexandrovsky city clinical hospital aged 52–76 with transitional cell carcinoma of the bladder. All examined patients were divided into five groups: the control group included conditionally healthy donors, while other patients were grouped according to their tumor stage (I, II, III and IV). Plasma plasminogen levels were determined by enzyme-linked immunosorbent assay, and the potential activity was measured by chromogenic plasminogen assay. Serine proteases fractions were obtained by the affinity chromatography method, and enzyme concentration in the selected fractions were determined by the Bradford method. Serine proteases distribution was investigated by electrophoresis in a polyacrylamide gel.ResultsIt was determined that the concentration, potential activity of plasminogen, and the total amount of serine proteases in plasma from BC patients were greater than the values of the corresponding indicators in healthy donors. This could be one of the factors contributing to increased proteolysis seen in the process of carcinogenesis. Plasminogen concentration in BC patients with stage IV disease; however, displayed a tendency to be reduced compared to earlier stages, and the potential activity of plasminogen was significantly lower in patients with stages III – IV BC. Futhermore, a tumor stage specific gradual decline in the serine protease plasma content was shown. The results of electrophoretic analysis established a significant diminishment in the percentage of high molecular weight components (under non-reducing conditions) and their complete disappearance (under reducing conditions) in plasma serine protease fractions from BC patients. A decline in the percentage of heavy and light plasmin chains in BC patients was also observed. Additionally, a rise in the degraded forms of plasminogen/plasmin content was seen in BC samples, as well as the presence of fractions corresponding to trypsin and NE (under non-reducing conditions) that were absent in the control samples.ConclusionThe results indicate significant changes in the proteolytic activity of plasma, from BC patients when compared to healthy controls, which is accompanied by alterations in serine protease distribution caused by tumor microenvironment pecularlities at the different stages of oncopathology