Ameliorative effect of ethanolic extract of Limnophila rugosa (Scrophulariaceae) in paracetamol- and carbon tetrachloride-induced hepatotoxicity in rats

Background Limnophila rugosa (Scrophulariaceae) is a perennial aquatic plant used as a diuretic and digestive tonic as well as in the treatment of diarrhea, dysentery, dyspepsia and urinary ailments. Genus Limnophila has been reported as hepatoprotective. The present study was undertaken to evaluate the hepatoprotective activity of the ethanolic extract of L. rugosa aerial part in paracetamol- and carbon tetrachloride-induced (CCl4) hepatotoxicity in albino Wistar rats. Ethanolic extract was subjected to high-performance liquid chromatography (HPLC) analysis for the estimation of phenolic and flavonoid compounds and gas chromatography–mass spectrometry (GC–MS) analysis for phytochemical analysis. The in vitro antioxidant activity was carried out by 2,2-diphenyl-1-picrylhydrazyl, nitric oxide radical and hydrogen peroxide assay. Hepatoprotective potential of L. rugosa was studied in paracetamol (750 mg/mg)- and CCl4 (1.25 ml/kg)-induced liver damage in albino rats at dose 200 and 300 mg/kg using silymarin (100 mg/kg) as standard. Lipid peroxidation, superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) were determined in liver tissue homogenate. Serum biochemical and histopathological examination was performed. Molecular docking analysis was performed to understand the molecular mechanism of hepatoprotective activity. Results HPLC analysis revealed predominance of rutin. GC–MS analysis revealed camphor as principal component. Ethanolic extract exhibited significant concentration-dependent scavenging efficacy. The altered biochemical chemical parameters: aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, bilirubin, cholesterol, albumin, globulin and total protein, were significantly improved at 200 and 300 mg/kg in experimental rats. Extract signified hepatoprotective by decreasing lipid peroxidation and upregulating SOD, CAT and GSH. The findings were well supported by histological analysis. 2-Butyl-2, 7-octadien-1-ol (-5.8) and camphor (-4.8) gave the highest docking score on the transforming growth factor-β1. Conclusions The ameliorative effect of L. rugosa in the rat model of hepatotoxicity could be attributed to its antioxidant potential and bioactive principles such as betulin, 5-hydroxy-6,7,4′-trimethoxyflavone (salvigenin), betulinic acid, ursolic acid, 3-octanol, acetophenone, anisylacetone, caryophyllene, cis-anethole and the compounds camphor and 2-butyl-2,7-octadien-1-ol identified from GC–MS analysis.

Glutathione detoxication processes in females suffering from sterility

Thirty females suffering from sterility were examined. Fertility disorders were accompanied by hyperandrogeny. Hormones FSH, LH, testosterone and progesterone in blood were studied. Besides, an activity of enzymes GPO, GTD, GST, G-6-PD in washed erythrocytes and tissue homogenate of ovary was investigated. Findings obtained demonstrate noticeable disorders in glutathione detoxication processes in females with infertility and they are related to hormones level, in particular androgens. An increase of GPO in females with hyperandrogeny showed deepupdates in metabolism processes of steroid hormones in ovary. It manifests beforehand for development of follicle that is predictive sign of unovulation in the beginning of menstrual cycle.

Influence of quercetin on morphological changes in rats testes after 180 days during central deprivation of luteinizing hormone

A relevant and popular area of research is the protective effect of the bioflavonoid quercetin, which makes it possible to use it to correct testicular dysfunction of various origins. The aim of the study was to determine the effect of quercetin on the microscopic organization of rat testicles, nitric oxide production and the intensity of oxidative stress in rat testicles on the 180th day of the experiment, during experimental central deprivation of luteinizing hormone synthesis caused by triptorelin solution. The experiment was performed on 20 adult male white rats. Rats were divided into 2 groups of 10 animals in each group: control group (I), group with central deprivation of LH synthesis + quercetin (II). Animals from the group with central deprivation of LH synthesis were injected subcutaneously with triptorelin acetate at a dose of 0.3 mg of active substance per kg and quercetin 100 mg per kg of body weight 3 times a week, while the control group was injected with saline. Ultrathin sections made by ultramicrotome were contrasted with 1 % aqueous uranyl acetate and lead citrate according to the Reynolds method and examined by electron microscopy. According to standard methods, the material was poured into paraffin blocks, from which sections with a thickness of 4 μm were made and stained with hematoxylin and eosin. Histological specimens were examined using a Вiorex 3 light microscope with a digital microfilter with software adapted for these studies. All biochemical studies were performed in 10 % of testicular tissue homogenate using a Ulab 101 spectrophotometer. Statistical processing of the study results was performed using Microsoft Office Excel software and Real Statistics 2019 extension. The nonparametric Mann-Whitney test was used to determine the statistical significance of differences between groups. Our study of the interstitial space in the testes of white rats showed the heterogeneity of populations of endocrinocytes and macrophages and the variability of structural and functional parameters. In the tissues of the testes in conditions of prolonged central deprivation of testosterone synthesis develops oxidative stress, which on the 180th day of the experiment leads to the development of edema of the interstitial space, followed by tissue fibrosis. Changes in the polarization of macrophages in our opinion may cause oxidative stress in the testes, as evidenced by increased iNOS activity and decreased arginase activity, but use of quercetin protects rat testicular tissue from oxidative damage caused by triptorelin by increasing direct antioxidant system and effects on the lipoxygenase pathway of arachidonic acid metabolism.

Pathogenicity of field strain of fowl aviadenovirus serotype 11 isolated from chickens with inclusion body hepatitis in Morocco

Outbreaks of inclusion body hepatitis have emerged in Morocco since 2013 and has resulted in significant economic losses to poultry farms. Three isolates of the causative virus, Fowl adenonovirus (FAdV)were characterized from chickens with IBH, but their pathogenicity has never been investigated. In this work, the pathogenicity of an isolate FAdV 11 (MOR300315 strain) was evaluated by inoculating a group of 40 SPF chickens at 3 days of age by oral route. A group of 40 chicks injected with phosphate-buffered saline solution was used as a control group. The infected chickens showed decreased weight gain from 3dpi. Necropsy displayed pallor and enlargement in liver, swelling and slight hemorrhage in kidney and spleen at 6 dpi. Histopathological changes were mainly characterized by severe and extensive hepatic necrosis associated with the presence of basophilic intra-nuclear inclusion bodies within hepatocytes. The FAdV was reisolated in chicken embryo fibroblast cell culture from liver tissue homogenate of infected chicken from 3 to 6 dpi. Viral DNA was detected by PCR in liver, kidney, spleen and cloacal swabs from 3 to 13 dpi. Antibody response against inoculated FAdV was appeared from 9 dpi. These results confirmed that the FAdV 11 strain is pathogenic in chicken. This study is the first experimental infection of FAdV 11 in chicken in Morocco, which increase our understanding of its pathogenicity in chickens and indicate that preventive measures against FAdV infection in poultry farms should be implemented in Morocco.

Thymoquinone Attenuates Immune Mediated Liver Injury Induced by Concanavalin A in Mice

Autoimmune hepatitis is an inflammatory disease and its incidence has been increasing. The features of hepatitis are the release of inflammatory cytokines, the elevation of AST and ALT, and hepatocyte apoptosis and necrosis. Concanavalin A considered as essential model represents the acute immune-mediated liver damage in rodents. Thymoquinone is well known herbal compound that exert hepatoprotective, anti-inflammatory, and antioxidant activity. In this study, we focus on the immunoregulatory and liver protective effect of thymoquinone in a mouse model of concanavalin A-induced liver injury. Twenty-four male mice were randomly divided into four groups each containing six animals: Negative control group, concanavalin A model group, thymoquinone 15mg/kg group, and thymoquinone 30mg/kg group. Blood was collected to detect the activities of alanine transaminase (ALT) and aspartate transaminase (AST) as well as liver tissue for the detection of tumor necrosis factor levels, after 8 hours of concanavalin A injection. Injecting mice with concanavalin resulted in a dramatic increase in serum level of both ALT and AST which indicate severe liver tissue damage with a significant increase in inflammatory cytokines TNF alpha - and INF levels, with a notable increase in NF-kB gene expression in mice liver tissue homogenate. Thymoquinone pretreatment revealed a dose-dependent increase in liver tissue protection. Conclusion: Thymoquinone has hepatoprotective and immune modulatory effects in concanavalin A induced immune mediated liver damage. Thymoquinone exerted its effect through attenuating NF and its downstream effectors TNF and INF in a dose dependent manner.

Emodin Promotes Autophagy and Prevents Apoptosis in Sepsis-Associated Encephalopathy through Activating BDNF/TrkB Signaling

<b><i>Objective:</i></b> Sepsis-associated encephalopathy (SAE) is a severe and common complication of sepsis and can induce cognitive dysfunction and apoptosis of neurons and neuroinflammation. Emodin has been confirmed to have anti-inflammatory effects. Thus, we sought to investigate the role of Emodin in SAE. <b><i>Methods:</i></b> The cecal ligation and puncture (CLP) method was used for the establishment of SAE in mice model. For treatment of Emodin, intraperitoneal injection of 20 mg/kg Emodin was performed before the surgery. The Morris water maze and open field tests were carried for measurement of cognitive dysfunction. Hematoxylin and eosin staining was for histological analysis of hippocampus. Cell apoptosis of hippocampus neurons was measured by TUNEL staining. Pro-inflammatory and anti-inflammatory cytokines in hippocampus tissue homogenate were evaluated by ELISA. BDNF/TrkB signaling-related proteins (TrkB, p-TrkB, and BDNF), autophagy-related proteins (LC3 II/I and Beclin-1), and apoptosis-related proteins (Bax, Bcl-2, and cleaved caspase-3) were detected by Western blotting. <b><i>Results:</i></b> Emodin significantly inhibited apoptosis and induced autophagy in hippocampal neurons of CLP-treated mice. In addition, Emodin significantly ameliorated CLP-induced cognitive dysfunction and pathological injury in mice. Meanwhile, Emodin notably inhibited CLP-induced inflammatory responses in mice via upregulation of BDNF/TrkB signaling, while the effect of Emodin was partially reversed in the presence of K252a (BDNF/TrkB signaling inhibitor). <b><i>Conclusion:</i></b> Emodin significantly inhibited the progression of SAE via mediation of BDNF/TrkB signaling. Thus, Emodin might serve as a new agent for SAE treatment.

FUNCTIONING OF NO-CYCLE IN THE SALIVA OF CHILDREN WITH TYPE 1 DIABETES MELLITUS

Introduction. The presence of type 1 diabetes mellitus in children is a major risk factor for periodontal disease. The aim of research work was to determine the activity of NO-synthase and arginase in saliva in children age with insulin-dependent diabetes mellitus. Materials and methods We examined 82 children including 56 children with type 1 diabetes mellitus and 26 children without somatic diseases. NO-synthase (NOS) activity was determined by the difference in nitrite concentration before and after incubation of tissue homogenate. Determining arginase activity was based on analysis the difference in the concentration of L-ornithine before and after incubation in phosphate buffer solution. Statistical processing was performed using Microsoft Office Excel. Research results and their discussion The violation of the indicators’ balance between groups 1 and 3 showed us a decrease in regenerative capacity in the mucous membrane in persons with type 1 diabetes mellitus. Increased ARG activity in group 4 children may lead to rivarly between NOS and ARG for L-arginine. Increased ARG activity in groups 2 and 4 compared with group 1 indicated an adaptive response aimed at repairing gum damage. Based on this, increased NO production from NOS is a consequence of insulin deficiency (systemic factor). Conclusions. The combination of systemic factor (type 1 diabetes mellitus) and local (chronic catarrhal gingivitis) leads to dysregulation of the NO-cycle and increasing of competition between NOS and ARG. ФУНКЦИОНИРОВАНИЕ NO-ЦИКЛА В СЛЮНЕ У ДЕТЕЙ С САХАРНЫМ ДИАБЕТОМ І ТИПА Кузь И.А., Акимов О.Е., Костенко В.А., Шешукова О.В., Максименко А.И., Писаренко Е.А. Полтавский государственный медицинский университет Вступление. Наличие у детей сахарного диабета І типа является основным фактором риска заболеваний пародонта. Целью исследования было определение активности NO-синтазы и аргиназы в слюне у детей с инсулинозависимым сахарным диабетом. Материалы и методы. Обследовано 82 ребенка, в том числе 56 детей с сахарным диабетом І типа и 26 детей без соматических заболеваний. Активность NO-синтазы (NOS) определяли по разнице в концентрации нитрита до и после инкубации гомогената ткани. Определение активности аргиназы основывалось на анализе разницы в концентрации L-орнитина до и после инкубации в фосфатном буферном растворе. Статистическая обработка проводилась с использованием Microsoft Office Excel. Результаты исследований и их обсуждение. Нарушение баланса показателей между 1 и 3 группами свидетельствовало о снижении регенерационной способности слизистой оболочки у лиц с сахарным диабетом І типа. Повышенная активность ARG у детей группы 4 может привести к неравенству между NOS и ARG для L-аргинина. Повышенная активность ARG в группах 2 и 4 по сравнению с группой 1 указала на адаптивный ответ, направленный на восстановление повреждений десен. Исходя из этого, повышенная продукция NO из NOS является следствием дефицита инсулина (системный фактор). Выводы. Сочетание системного фактора (сахарный диабет І типа) и местного (хронический катаральный гингивит) приводит к нарушению регуляции NO-цикла и усилению конкуренции между NOS и ARG.

Evaluation of Paromomycin Loaded Chitosan Nanoparticles and Oxidative Stress Activities against Entamoeba Histolytica

This investigation was carried out to estimate the antiparasitic potential of chitosan nanoparticles loaded with paromomycin against Entamoeba histolytica infected. After rats inoculated orally in a dose 103 viable cysts for acute infection; then treated with paromomycin, chitosan nanoparticles as a single or combined therapy given for seven days. Stool examination revealed a significant decrease in the number of Entamoeba histolytica cysts in all treated infected rats compared with infected non-treated. Combined treatment provided better results than single treatment. The best effect was observed in the group of rats treated with chitosan nanoparticles loaded with paromomycin. Also, the oxidative stress markers Glutathione (GSH) and Lipid Peroxide (Malondialdehyde) (MDA) were assessed in liver tissue homogenate. The current work is the first time of using chitosan nanoparticles loaded with paromomycin as therapeutic agents against experimental amoebiasis. It was shown that the highest degree of effectiveness attained by the synergistic action of paromomycin and chitosan nanoparticles as was indicated by lower parasite count and GSH, MDA concentration.

Experimental Study of the Effect of Serum Hydrogen Sulfide on the Course of the Inflammatory Process in the Vaginal Wall

Introduction: Various pathological conditions are characterized by the influence of hydrogen sulfide level on the course of the pathological process. This study examines the effect of serum hydrogen sulfide levels on the inflammatory process in the vaginal wall of rats. Aims: To evaluate the effect of excess and deficiency of serum hydrogen sulfide on the course of the inflammatory process in the vaginal wall of rats. Methodology: The study was performed on 125 female Wistar rats under 1 year of age and weighing 160.0 to 200.0 grams. All animals were divided into 7 groups: control (intact rats) and 6 experimental groups with different H2S levels and different treatment approaches of inflammation in the vaginal wall. The level of serum hydrogen sulfide was studied and the levels of TNF-α and IL-1β in the tissue homogenate of the vaginal wall were determined. In all experimental groups, the study was performed in dynamics - 10 min, 4, 8 and 24h after simulation of inflammation. Results: The dynamics of local levels of TNF-α and IL-1β in all groups had a similar trend and was characterized by the rapid development of the inflammatory process from its simulation to 4 hours of study, followed by gradual attenuation of inflammation and almost complete normalization of the studied indicators for 24 hours. Preliminary serial introduction of sodium hydrosulfide, as a donor of hydrogen sulfide, allowed to reduce the degree of manifestation of the inflammatory process and to achieve faster normalization of the studied parameters. At the same time, the artificially created deficiency of serum hydrogen sulfide (previous serial administration of propargylglycine) prolonged the duration and increased the studied indicators of inflammation in the vaginal wall. Conclusions: The course and intensity of the inflammatory process in the vaginal wall of rats are directly dependent on the background level of serum hydrogen sulfide.

Geniposide Ameliorates Liver Fibrosis Through Reducing Oxidative Stress and Inflammatory Respose, Inhibiting Apoptosis and Modulating Overall Metabolism

Liver fibrosis is a progressive liver damage condition caused by various factors and may progress toward liver cirrhosis, and even hepatocellular carcinoma. Many studies have found that the disfunction in metabolism could contribute to the development of liver fibrosis. Geniposide, derived from Gardenia jasminoides J. Ellis, has been demonstrated with therapeutic effects on liver fibrosis. However, the exact molecular mechanisms of such liver-protection remain largely unknown. The aim of this study was to explored the effect of geniposide on metabolic regulations in liver fibrosis. We used carbon tetrachloride (CCl4) to construct a mouse model of liver fibrosis and subsequently administered geniposide treatment. Therapeutic effects of geniposide on liver fibrosis were accessed through measuring the levels of hepatic enzymes in serum and the pathological changes in liver. We also investigated the effects of geniposide on inflammatory response, oxidative stress and apoptosis in liver. Furthermore, serum untargeted metabolomics were used to explore the metabolic regulatory mechanisms behind geniposide on liver fibrosis. Our results demonstrated that geniposide could reduce the levels of hepatic enzymes in serum and ameliorate the pathological changes in liver fibrosis mice. Geniposide enhanced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and decreased methane dicarboxylic aldehyde (MDA) levels in liver. Geniposide treatment also decreased the levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor-alpha (TNF-a) in liver tissue homogenate. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining demonstrated that geniposide could reduce the apoptosis of hepatocytes. Geniposide increased the protein expression of B-cell lymphoma-2 (Bcl-2) and downregulated the protein expression of Bcl-2 Associated X (Bax), cleaved-Caspase 3, and cleaved-Caspase 9. Serum untargeted metabolomics analysis demonstrated that geniposide treatment improved the metabolic disorders including glycerophospholipid metabolism, arginine and proline metabolism, and arachidonic acid (AA) metabolism. In conclusion, our study demonstrated the protective effects of geniposide on liver fibrosis. We found that geniposide could treat liver fibrosis by inhibiting oxidative stress, reducing inflammatory response and apoptosis in the liver, and modulating glycerophospholipid, and arginine, proline, and AA metabolism processes.