Proinflammatory Cytokine Gene Expression Research Articles

The nanocomposite of chitosan-vitamin C modulates the expression of immune and antioxidant-related genes in Nile tilapia stressed with lead (Pb)

Fish cultivated in a polluted environment usually face complicated disease cases. Pollution is an evitable event in aquaculture that weakens surface immunity, delaying and lowering the immune responses of aquatic animals. In an indoor experiment, Nile tilapia were reared in water polluted with lead (Pb) at a concentration of 51 µg/l for 8 weeks. A trial was conducted to mitigate the immunosuppression using pre-acclimatized Nile tilapia that were subdivided into six groups in which the exposed fish treated with nanocomposite (chitosan-vitamin C-protected nanoparticles (CVC-NPs)) consisted of 1 g of chitosan nanoparticles containing 400 mg vitamin C/kg fish and 200 mg vitamin C/kg fish feed for 1 week. Fish exposed to Pb pollution were immunocompromised as gene expression of immune-related cytokine and antioxidant enzymes significantly declined. Innate immunity was drastically impacted as OBA, phagocytosis and serum antibacterial activity (SAA) showed low values compared to the control fish; meanwhile, exposed fish was challenged against Streptococcus agalactiae and showed a high mortality rate (MR%). Dietary nanocomposite could restore normal immune status, and supplemented fish achieved low MR% with significantly high gene expression of proinflammatory cytokines. It was concluded that adding 200 mg of CVC-NPs/kg fish feed for 1 week could effectively protect Nile tilapia from S. agalactiae, having superiority over the high dose of 400 mg of CVC-NPs.

Sleeve gastrectomy and pancreatic omentoplasty decrease insulin resistance: Impact on pro-inflammatory and anti-inflammatory cytokine gene expression

Sleeve gastrectomy and pancreatic omentoplasty decrease insulin resistance: Impact on pro-inflammatory and anti-inflammatory cytokine gene expression

Evaluation of cellular immune response in rabbits after exposure to cobra venom and purified toxin fraction

Snakebite by a cobra is considered neurotoxic as the cause of neuromuscular paralysis mediated by low molecular weight toxins, which are major toxin components of cobra. However, these toxins represent a problem in generating antibodies owing to their low immunogenicity. Developing complementary strategies to improve the antibody response could be a useful approach to creating better therapeutic antivenoms with higher neutralizing potencies. To develop simple immunization strategies for more potent antivenoms by studying the effects of combining crude cobra venom and toxin fraction in a complementary way. The evaluation of specific cell immunology and cytokine mediators for relevant immune responses will be measured in a rabbit model using four simple immunization strategies. Flow cytometry will be used to quantify the number of B and T cells, and qRT-PCR will be used to ascertain the cytokine genes expressed. B cells with anti-CD20 were seen on D14, and a booster dose was insufficient to maximize the antibodies. Conversely, anti-CD5 for T cells decreased periodically but remained stable. Using a mixture of crude cobra venom and its <10 kDa fraction, peak expression of pro-inflammatory cytokine genes was seen in D42 or D58, with a rise of 4 and 6 folds. Similarly, gene expression of pro-inflammatory cytokines was greater than that of anti-inflammatory cytokines (IL-4 and IL-10), which were up-regulated after D42. Thus, immunization with both the crude and its <10 kDa fraction of cobra venom seems to have synergistic effects that boost cytokines, activate the immune system, and cause lymphocyte differentiation.

Fractionated alpha and mixed beam radiation promote stronger pro-inflammatory effects compared to acute exposure and trigger phagocytosis

Introduction and methodsAiming to evaluate safety aspects of a recently proposed approach to target Alzheimer’s disease, we mimicked a complex boron neutron capture therapy field using a mixed beam consisting of high- and low-linear energy transfer (LET) radiation, 241Am alpha particles (α) and/or X-ray radiation respectively, in human microglial (HMC3) cells.ResultsAcute exposure to 2 Gy X-rays induced the strongest response in the formation of γH2AX foci 30 min post irradiation, while α- and mixed beam-induced damage (α:X-ray = 3:1) sustained longer. Fractionation of the same total dose (0.4 Gy daily) induced a similar number of γH2AX foci as after acute radiation, however, α- or mixed irradiation caused a higher expression of DNA damage response genes CDKN1A and MDM2 24 h after the last fraction, as well as a stronger decrease in cell viability and clonogenic survival compared to acute exposure. Phosphorylation of STING, followed by phosphorylation of NF-κB subunit p65, was rapidly induced (1 or 3 h, respectively) after the last fraction by all radiation qualities. This led to IL-1β secretion into the medium, strongly elevated expression of pro-inflammatory cytokine genes and enhanced phagocytosis after fractionated exposure to α- and mixed beam-irradiation compared to their acute counterparts 24 h post-irradiation. Nevertheless, all inflammatory changes were returning to basal levels or below 10–14 days post irradiation.DiscussionIn conclusion, we demonstrate strong transient pro-inflammatory induction by daily high-LET radiation in a microglia model, triggering phagocytosis which may aid in clearing amyloid beta, but importantly, from a safety perspective, without long-term alterations.

The ameliorative effect of pioglitazone against colistin-induced nephrotoxicity is mediated by inhibition of NF-κB and restoration of Nrf2 signaling: An integrative bioinformatics prediction-guided in vitro study.

Pioglitazone, an anti-diabetic drug, has been previously shown to ameliorate kidney damage through anti-inflammatory and antioxidant effects. In this study, we employed an integrative bioinformatics approach to study the possible mechanisms involved in the mitigative effect of pioglitazone against colistin-induced nephrotoxicity. Next, we validated the results obtained from the bioinformatics study by pre-treating human kidney-2 (HK-2) cell line with pioglitazone 100 μM for 30 minutes and then treating the cells with colistin sulfate 1200 μM for 24 hours. Inflammatory signaling by cytokines and the nuclear factor erythroid 2 related factor 2 (Nrf2) signaling pathways were predicted to be involved in the ameliorative effect of pioglitazone against colistin-induced nephrotoxicity. The nuclear factor kappa B subunit p65 (NF-κB p65) and Nrf2 were among the predicted transcription factors regulating the hub genes. Moreover, miR-24, miR-16, and miR-21 were identified as potential pathogenic miRNAs regulating the hub genes. In contrast, miR-17, miR-27a, and miR-146a were identified as potential protective miRNAs. The in vitro study indicated that pioglitazone pre-treatment increased cell viability in HK-2 cells exposed to colistin. Pioglitazone pre-treatment reduced the expression of pro-inflammatory cytokine genes (IL6 and TNF). Moreover, pioglitazone reduced the protein expression of NF-κB p65 and increased the protein expression of Nrf2. The protective effect of pioglitazone against colistin-induced toxicity in HK-2 cells is related to its anti-inflammatory and antioxidant activity through modulating NF-κB-mediated inflammatory signaling and Nrf2-mediated antioxidative stress signaling.

Dynamic Changes in the Intraepithelial Lymphocyte Numbers Following Salmonella Typhimurium Infection in Broiler Chickens.

At day 21 of age, Ross-308 broilers were orally gavaged with 7.5 × 106 CFU/mL S. Typhimurium (n = 30), and another 30 birds were kept as the control. The body weight of birds was recorded on days 0, 2, 7, and 14 days post-infection (dpi) to calculate body weight gains (BWGs). At each time point, seven birds per group were euthanized for sample collection to acquire IELs and lymphocytes from the ileum and spleen for flow cytometric analysis. A reduction in BWGs of the infected groups compared to the control group was observed only at 2 dpi. Additionally, there were no changes in the expression of IFN-γ, IL-1β, and TNF-α in the ileum at 2 and 7 dpi. The number of IELs increased significantly following Salmonella infection in the ileum at 2 and 7 dpi without any changes in spleen lymphocytes. The increase in the total number of IELs was derived from the elevated numbers of conventional CD8αβ+TCRαβ+ and natural IEL populations (CD4-CD8-TCRαβ+, CD8αα+TCRαβ+, TCRγδ+, non-T cells (TCRneg, and iCD8α cells)). The increase in regulatory IELs and the stable expression of proinflammatory cytokine genes during the first week of infection suggests the potential role of IELs in modulating intestinal inflammation.

Anti-inflammatory effects of quercetin in lipopolysaccharide-activated cortical astrocytes

Dysregulated neuroinflammation is a hallmark of multiple neurodegenerative diseases, characterized by increased pro-inflammatory cytokines, and reduced anti-inflammatory cytokines, and is mediated by resident astrocytes. Previous studies have demonstrated the neuroprotective effects of quercetin hydrate, a plant flavonoid, however, limited studies exist showing the targets of quercetin in astrocytes. Here, we demonstrate the anti-inflammatory effects of quercetin hydrate at the gene expression level and the involvement of signalling pathway molecules. The rat primary cortical astrocytes were preincubated with different concentrations of quercetin (25–200 μM) followed by lipopolysaccharide (LPS) stimulation. Semi-quantitative PCR analysis showed the therapeutic potential of quercetin by decreasing the gene expression of pro-inflammatory cytokines (IL-1β, IL-6), and Cyclooxygenase-2, and increasing Hemeoxygenase-1 in LPS-stimulated astrocytes. By Immunoblotting analysis, we document the modulatory effects of quercetin on phospho p38 and phospho ERK1/2 in LPS-stimulated astrocytes. The study indicates the anti-inflammatory effects of quercetin against inflammatory responses in astrocytes.

The effect of hyperbaric oxygen therapy on oxidative stress and inflammation in patients with diabetic foot ulcers: A Preliminary Study

IntroductionPatients with an uncontrolled glycemic index develop a wide variety of pathologies associated with diabetes, such as diabetic foot ulcers (DFUs). Hyperbaric oxygen therapy (HBOT) is an adjunctive therapy used to heal wounds and prevent lower extremity amputations in this population. ObjectiveThis preliminary study aimed to evaluate how HBOT impacts inflammation in patients with Wagner stages 2-4 DFUs by analyzing its effect on the gene expression of key oxidative stress regulators SOD1, SOD2, and GPX2, of pro-inflammatory cytokines TNFα, IL-1β, IL-4, and IL-12, and of the NLRP3 inflammasome. MethodsThe effect of HBOT was assessed in 15 patients with Wagner stages 2-4 DFUs that underwent 30 sessions in the hyperbaric chamber. This protocol is registered on Clinical Trials under the title Hyperbaric Oxygen Therapy in Diabetic Foot (15/07/2024) with the number NCT06502808. Blood samples were collected, and relative gene expression was assessed by quantitative real-time polymerase chain reaction (qPCR). ResultsThe hyperbaric chamber treatment increased the expression of SOD1 and GPX2 genes (0.4 and 3 times, respectively) after 30 sessions compared to baseline levels. Similarly, the gene expression of pro-inflammatory cytokines IL-1β, IL-12, IL-4, and NLRP3 increased after 30 sessions (2.1, 0.4, 1.5, and 1.2, respectively), while the expression of the TNFα gene decreased (0.5 times). Clinically, the patients’ lesions were fully resolved. ConclusionsHBOT directly influences the gene expression of several potent antioxidants and pro-inflammatory cytokines, thus favoring angiogenesis and blood circulation in the extremities.

Pharmacotherapeutic potential of bilobetin to combat chromium induced hepatotoxicity via regulating TLR-4, Nrf-2/Keap-1, JAK1/STAT3 and NF-κB pathway: A pharmacokinetic and molecular dynamic approach

BackgroundChromium (Cr) is one of the top-notch noxious heavy metals that is documented to exert deleterious effects on various body organs including the liver. Bilobetin (BLB) is a natural flavonoid which exhibits a wide range of medicinal properties. AimThis trial was executed to investigate the pharmacotherapeutic potential of BLB to avert Cr instigated hepatotoxicity via modulating TLR4, JAK1/STAT3, Nrf-2/Keap-1 and NF-κB pathway. Research layoutOur trial was executed on thirty-six male albino rats that were segregated into four equal groups including the control, Cr (10 mg/kg), Cr (10 mg/kg) + BLB (12 mg/kg) and BLB (12 mg/kg) alone treated group. Various biochemical parameters were assessed by using qRT-PCR, molecular docking, molecular dynamic simulation and histological approaches. FindingsOur results revealed that Cr administration significantly impaired the health of hepatic tissues by reducing the gene expression of Nrf-2 and its downregulating genes while promoting the levels of oxidative stress markers (ROS and MDA). Moreover, Cr administration upregulated the hepatic enzymes including ALT, GGT, AST, and ALP while concurrently decreasing the levels of total protein and albumin. Cr exposure also elevated the gene expression of pro-inflammatory cytokines including toll-like receptor 4 (TLR4), high mobility group box 1 (HMGB1) nuclear factor kappa B (NF-κB), Janus kinase 1 (JAK1), signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor alpha (TNF-α), C-reactive proteins, interferon-gamma inducible protein-10 (IP-10), Interleukin beta-1(IL-1β), monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2). Hepatic apoptosis was observed to be elevated following the Cr intoxication. Nonetheless, BLB treatment remarkably alleviated the hepatic damages via regulating the biochemical as well as histological profile of liver. Our findings are further endorsed by molecular docking analysis that demonstrated that BLB exhibit strong binding affinity to Keap-1 and STAT3 thus supporting its efficient hepatoprotective potential. ConclusionBLB protected the hepatic tissues via regulating Cr induced impairments. These findings were confirmed by molecular docking and molecular dynamic simulation analysis.

Expression of key cytokines in dog macrophages infected by Leishmania tarentolae opening new avenues for the protection against Leishmania infantum

The detection of Leishmania tarentolae in sympatric areas where Leishmania infantum is endemic raised questions regarding the protective effect exerted in dogs by L. tarentolae when in coinfection. This study aimed monitoring the in vitro gene expression of pro- (IFN- γ; TNF-α; IL-12) and anti-inflammatory (IL-4; IL-6; IL-10) cytokines in primary canine macrophages infected by L. tarentolae and L. infantum in single and in co-infections. Macrophages differentiated from dog blood mononuclear cells were infected with the L. tarentolae field-isolated (RI-325) and laboratory (LEM-124) strains, with L. infantum laboratory strain (IPT1), or both. Infection and the number of amastigotes per infected cell were evaluated microscopically by counting a total of 200 cells between 4 and 96 h. Cytokine gene expression was analyzed by real-time PCR from infected macrophages mRNA. Single infections presented higher expression of the cytokines IL-4 and IL-6, and lower of IL-12. Co-infections induced a lower gene expression of IL-4 and IL-6, and a higher gene expression of IL-12, correlating with the low amastigote burden despite the slight increase of infected cells. Data highlight the potential protective effect of L. tarentolae against L. infantum in co-infection by the reduced anti-inflammatory and increased pro-inflammatory cytokines gene expression, opening new perspectives for a canine vaccine development exploiting the non-pathogenic L. tarentolae.

Modulating the anti-inflammatory and barrier protective effects on intestinal epithelial cells utilizing an aqueous Aspalathus linearis extract in vitro

BackgroundRooibos is a natural herbal plant containing numerous unique polyphenols which have been associated with certain health benefits. Several of these bioactive constituents have been linked to promoting gut health through its beneficial actions such as anti-inflammatory and anti-spasmodic characteristics. PurposeThe study aims to compare the anti-inflammatory and barrier protective effects of an unfermented and fermented rooibos aqueous extract on intestinal porcine epithelial cells (IPEC-J2). MethodsAqueous extracts of unfermented and fermented rooibos were prepared, and chemical and antioxidant analysis were performed of each extract. Intestinal porcine epithelial cells (IPEC-J2) were pre-treated with either unfermented or fermented aqueous rooibos extracts (0.1 and 0.05 mg/ml) then exposed to 10 µg/ml of Escherichia coli lipopolysaccharide (LPS). Cell viability, protein and gene expression of pro-inflammatory cytokines were investigated to determine the anti-inflammatory effect of both extracts. Barrier integrity of IPEC-J2 was investigated by measuring the Transepithelial electrical resistance (TEER) and quantifying tight junction gene expression. ResultsBoth the unfermented and fermented rooibos extracts have significant anti-inflammatory properties by reducing pro-inflammatory cytokine production. Interestingly for certain genes (IL-8, IL-6, IL-1B and CCL20) the fermented extracts, specifically at 0.1 mg/ml, exhibited greater effectiveness by decreasing the expression after LPS-induced inflammation whilst for another gene (TNF-α) the unfermented extract at both concentrations was more efficient in decreasing expression and ultimately protecting the IPEC-J2 cells from inflammation. When evaluating the barrier integrity, 0.1 mg/ml of the unfermented rooibos extract showed the most significant barrier protective effects through upregulation of tight junction proteins, ZO-1, Occludin as well as Claudin-4. ConclusionHigher concentration of the fermented rooibos extract prevented the inflammatory response significantly whilst the higher concentration of the unfermented rooibos extract enhanced tight junction expression and protected barrier integrity in LPS-induced inflamed intestinal cells. This is attributed to the differences in the polyphenol content of each extract and demonstrating the unique activity mechanisms of different rooibos extracts to reduce onset of early inflammation.

PNSC5325 prevents acute respiratory distress syndrome by alleviating inflammation and inhibiting extracellular matrix degradation of alveolar macrophages

BackgroundAcute respiratory distress syndrome (ARDS) is characterized by severe inflammation and significant extracellular matrix (ECM) degradation in the lungs. Our prior research identified the CtBP2-p300-NF-κB (C-terminal-binding protein 2-histone acetyltransferase p300-nuclear factor kappa B) transcriptional complex as critical in ARDS by activating pro-inflammatory cytokine genes. MethodsAn ARDS mouse model was established using intratracheal instillation of lipopolysaccharide (LPS). Small molecules that inhibit the CtBP2-p300 interaction were identified through AlphaScreen. RNA sequencing (RNA-Seq) was conducted to determine differential gene expression. Immunoprecipitation and co-immunoprecipitation analyzed protein interactions. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunoblotting detected gene and protein expression. Histological staining evaluated tissue damage. ResultsThrough AlphaScreen, two natural compounds, PNSC2477 and PNSC5325, were identified for their ability to inhibit the CtBP2-p300 interaction. While PNSC2477 demonstrated toxicity and was deemed unsuitable for further research, PNSC5325 exhibited minimal toxicity. PNSC5325 effectively inhibited the CtBP2-p300 interaction and reduced pro-inflammatory cytokine gene expression. RNA-Seq analysis of PNSC5325-treated cells indicated significant suppression of pro-inflammatory cytokine genes and matrix metalloproteinases (MMPs). Further molecular studies revealed that the CtBP2-p300 complex, in conjunction with activator protein 1 (AP1), activates MMP expression. PNSC5325 simultaneously suppressed both pro-inflammatory cytokines and MMPs by targeting the CtBP2-p300 complex. In LPS-injected mice, PNSC5325 administration significantly reduced ARDS incidence by inhibiting inflammatory and MMP genes. ConclusionThese findings suggest that PNSC5325 protects against ARDS by inhibiting key inflammatory and ECM degradation pathways, highlighting its potential as a novel therapeutic agent for ARDS and paving the way for further clinical investigations.

Effects of Conditioned Media From Human Umbilical Cord-Derived Mesenchymal Stem Cells on Tenocytes From Degenerative Rotator Cuff Tears in an Interleukin 1β-Induced Tendinopathic Condition.

Evidence suggests that mesenchymal stem cells (MSCs) are safe for treating different tendinopathies. Synovial fluid is a pooled environment of biomarkers from the inflammatory and degenerative joint cavity. Understanding the effects of synovial fluid on MSCs is important, as it is the first microenvironment that administered MSCs encounter. Several studies have reported that exposure to osteoarthritic synovial fluid-activated MSCs increased the release of soluble factors; however, the paracrine effects of shoulder synovial fluid-stimulated umbilical cord-derived MSCs (SF-UC-MSCs) on tendinopathy have yet to be investigated. To assess the effects of the conditioned media from SF-UC-MSCs on tenocytes from degenerative rotator cuff tears in an interleukin-1β (IL-1β)-induced tendinopathic condition. Controlled laboratory study. UC-MSCs were isolated and cultured from healthy, full-term deliveries by cesarean section. Tenocytes were isolated and cultured from patients with degenerative rotator cuff tears. Conditioned media were obtained from UC-MSCs stimulated with synovial fluid. To evaluate the gene expression of proinflammatory and anti-inflammatory cytokines, enzymes and their inhibitors, matrix molecules, and growth factors, the tenocytes were cultured with IL-1β and 50% of the conditioned media from the SF-UC-MSCs; quantitative, real-time, reverse transcriptase polymerase chain reaction was also performed. A prostaglandin E2 (PGE2) assay was performed to investigate the PGE2 level secreted by the tenocytes. Western blotting was performed to examine protein synthesis of collagen type I and III. Cell viability, senescence, and apoptosis assays were also performed. The conditioned media from the SF-UC-MSCs interfered with the inflammatory gene expression on tenocytes induced by IL-1β, but it increased the gene expression of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-3. Meanwhile, the conditioned media decreased the PGE2 level on cells induced by IL-1β. It did increase the type I/III ratio of gene expression and protein synthesis, mainly through the induction of type I collagen. Conditioned media of SF-UC-MSCs reversed senescence and apoptosis induced by IL-1β. Study findings indicated that the conditioned media from SF-UC-MSCs had anti-inflammatory effects and cytoprotective effects on IL-1β-treated tenocytes from degenerative rotator cuff tears. UC-MSCs have useful potential for the treatment of tendinopathy in practice.

Features of Brain Damage after Neutron Irradiation of the Head and Modification of the Damage by Lactoferrin.

: Mouse heads were irradiated in a beam of neutrons and gamma rays from the IR-8 nuclear reactor. The brain cells of control and irradiated mice were isolated using Percoll. Neurons and resting and activated microglial cells were analyzed using the fluorescently labeled antibodies and flow cytometry. The level of DNA double-strand breaks in neurons was determined by γH2AX histone content. Cytokine gene expression in the hippocampus was studied by RT-PCR. Behavior and cognitive functions were studied using the open field, Morris water maze, and novel object recognition tests. LF was isolated from female colostrum by preparative ion-exchange chromatography and purified by affinity chromatography on heparin-Sepharose. : γ,n-Irradiation of the mouse head at a dose of 1.5 Gy led to an increase in the level of DNA double-strand breaks in neurons. Twenty-four hours after irradiation the total number of cells and the number of neurons in the isolated fraction of brain cells decreased, but the number of microglial cells remained unchanged. The number of resting and activated microglia did not change within 3-72 h after γ,n-irradiation. The expression level of the TNFα, IL-1β, and IL-6 genes increased 2 months after γ,n-irradiation of the mouse head at a dose of 1.5 Gy, indicating the development of neuroinflammation. At this time, irradiated mice demonstrated the anxiety-like behavior and impaired spatial and episodic memory. A single i.p. administration of human LF to mice immediately after γ,n-irradiation of the head did not affect the observed radiation-induced disturbances, but decreased the gene expression levels of TNFα, IL-1β, and IL-6 pro-inflammatory cytokines and increased the gene expression level of TGFβ anti-inflammatory cytokine in the hippocampus 2 months after radiation exposure. The obtained results indicate a partial decrease in the level of hippocampal neuroinflammation of irradiated animals treated with LF. . γ,n-Irradiation of the mouse head at a dose of 1.5 Gy leads to DNA damage of neurons and the decrease in the number of neurons. Microglia cells are more resistant to such radiation exposure. Late after head-only γ,n-irradiation, mice develop neuroinflammation, which is detected by an increase in the pro-inflammatory cytokine gene expression in the hippocampus and also by anxiety-like behavior and impaired cognitive functions. A single LF administration leads to a partial decrease in the neuroinflammation level, but does not affect the other studied parameters. The optimal dosing regimen of LF remains to be determined to preserve cognitive functions after γ,n-irradiation of the brain.

SBP1 contributes to mesangial proliferation and inflammation through mitochondrial respiration in glomerulus during IgA nephropathy

Mesangial expansion and proliferation have been implicated in the pathogenesis of IgA nephropathy (IgAN). Mesangial cells in glomerulus are important contributors to commencement of IgAN. From minimal mesangial expansion to diffuse proliferation, the mesangial alteration is linked to clinical and pathological features of IgAN. Although selenium-binding protein 1 (SBP1) is associated with tissue injury, the roles of SBP1 in mesangial proliferation and inflammation in glomerulus during IgAN remains unclear. In the present study, we found that SBP1 gene levels were elevated in kidney tissues of patients with IgAN. Also, SBP1 protein levels were elevated in proliferative mesangial cells of glomerulus in kidney tissues from patients with IgAN. Urinary SBP1 protein levels were elevated in patients with IgAN. Elevated urinary SBP1 levels were positively correlated with segmental glomerulosclerosis of the Oxford classification related to mesangial proliferation in patients with IgAN. Over-expression of SBP1 induced cellular proliferation via mitochondrial respiration in human renal mesangial cells. Consistently, SBP1 knockdown and mitochondrial respiration inhibition suppressed cellular proliferation and induced mitochondrial oxidative stress in human renal mesangial cells. Furthermore, SBP1 induced pro-inflammatory phenotype by gene expression and production of pro-inflammatory cytokines and chemokines including IL-6, CXCL10, and CCL5 via NF-κB activation in human renal mesangial cells. These results suggest that SBP1 contributes to mesangial proliferation and inflammation via mitochondrial respiration during IgAN.

Phytochemical profiling and biological evaluation of the residues from industrial hemp (Cannabis sativa L.) inflorescences trimming: Focus on water extract

The study focused on investigating the phytochemical composition and bio-pharmacological properties of a water extract obtained from industrial hemp by-products, after trimming female inflorescences of Cannabis sativa L., cultivar Kompolti. The extract was found to be rich in phenolic compounds, particularly gallic acid, catechin, coumaric acid, ferulic acid, and benzoic acid. It also contained significant amounts of cannabidiolic acid and cannabigerolic acid as the main terpenophenols. To assess the biological potential of the extract, various ecotoxicological assays were conducted. The extract did not show significant phytotoxic effects on seedling germination in the allelopathic assay. In the brine shrimp (Artemia salina) lethality test, the LC50 value was 1.726 mg/mL, and in the Daphnia magna test, the extract showed no cardiotoxicity effects at the same concentration; thus, confirming its biocompatibility with these eukaryotic organisms. Additionally, the extract did not induce cytotoxicity in the murine C2C12 cell line till to the concentration of 1000 μg/mL. In isolated mouse prostate specimens, the extract (10–1000 μg/mL) also prevented LPS-induced gene expression of pro-inflammatory cytokines, namely tumor necrosis factor (TNF)α, interleukin (IL)-6, and IL-1β. Furthermore, the extract exhibited inhibitory effects on the growth of pathogenic bacterial, fungal, and dermatophyte species commonly associated with prostatitis. These results suggest that the residual powder from female inflorescence trimming can be considered an innovative plant material from the industrial hemp supply chain that deserves to be valorised in terms of recycling and upcycling; thus, reducing the environmental impact of the by-products and also opening the way for innovative health-promoting agents.

Skeletal myocyte-specific knockout of Ptpn1 and Ptpn2 enhances inflammatory response in muscle and increases mortality in polymicrobial sepsis

Abstract Background Critically ill patients often suffer from heart and skeletal muscle atrophy and failure (intensive care unit acquired weakness; ICUAW). Inflammation and sepsis are major risk factors for ICUAW. Critically ill patients with sepsis frequently develop insulin resistance that decreases protein synthesis and increases protein degradation in muscle eventually leading to ICUAW. The link between inflammation and insulin resistance is not well understood, but protein tyrosine phosphatases (PTP), that inhibit insulin signaling via insulin receptor (INSR) dephosphorylation as well as inflammation-associated pathways, are implicated. Hypothesis: We hypothesized that PTP1B and TC-PTP mediate inflammation-induced insulin resistance and muscle atrophy in heart and skeletal muscle. Methods and Results qRT-PCR analyses showed that the proinflammatory cytokines tumor necrosis factor (TNF), interleukin (IL)-1β, and IL-6 caused a ~2-fold increase in Ptpn1/PTP1B and Ptpn2/TC-PTP mRNA expression in C2C12 myocytes as well as in neonatal rat ventricular cardiomyocytes. Inhibition of the IL-6/GP130 pathway by siRNA and inhibitors in myocytes in vitro and skeletal muscle specific gp130 knockout mice in vivo attenuated IL-6-induced Ptpn1/PTP1B and Ptpn2/TC-PTP mRNA expression. CLP-induced polymicrobial sepsis was associated with a significant increase in gene expression and protein content of pro-inflammatory cytokines (Tnf, Il1b, Ifng) and leukocyte markers (CD68, CD11c, F4/80) in muscle of skeletal myocyte specific double knockout (Ptpn1+Ptpn2loxP/loxP, MCK cre, dKO) compared to wildtype (Ptpn1+Ptpn2loxP/loxP) mice in sepsis. This also resulted in a lower survival rate of dKO mice in sepsis (42% WT vs. 23% dKO). Immunohistological analyses revealed centralized nuclei, macrophage/monocyte infiltration and enhanced regeneration (Myh3, Pax7, Ki67) in tibialis anterior muscle of septic dKO mice. A strong activation of the NLRP3-inflammasome, as indicated by cleavage of caspase 1, GSDMD, GSDME and caspase 8, indicative for a pronounced inflammation and activated cell death pathways was observed in muscle of septic dKO mice. Conclusion The proinflammatory cytokine IL-6 increases the expression of Ptpn1 and Ptpn2 and enhances overall PTP activity. Selective inhibition of the IL-6R/GP130 signaling pathway by JAK2- and STAT3-inhibition as well as Gp130 deletion in myocytes in vivo attenuates those effects. Muscle-specific deletion of Ptpn1 and Ptpn2 in skeletal muscle leads to increased mortality, enhanced pro-inflammatory response and induced inflammatory, programmed cell death pathways in sepsis. Our data show that Ptpn1 and Ptpn2 play a key anti-inflammatory role in skeletal muscle.

Differential Expression of Mitogen-Activated Protein Kinase Kinase 3 (MAP2K3) and Interleukin12B (IL12B) in the Bursa of Fabricius of chicken infected with Infectious Bursal Disease Virus

Infectious Bursal Disease (IBD) caused by Infectious Bursal Disease virus (IBDV), poses a significant threat to the poultry industry, causing immunosuppression and economic losses since its discovery in 1957. To understand the host response to IBDV infection, this study examined the modulation of MAP2K3 and IL12B in chickens infected with IBDV. A local IBDV isolate propagated in embryonated chicken eggs was inoculated into 18-day-old chicken. The mRNA transcription levels of MAP2K3 and IL12B in bursal tissue were quantified by real-time quantitative polymerase chain reaction on day two post-infection and compared with those of uninfected, age-matched control birds (sham inoculated). The study revealed increased expression of the MAP2K3 and IL12B genes, indicating their role in inflammatory response during IBDV infection. Thus, the present investigation sheds light on the involvement of MAP2K3 and IL12B in the inflammatory response to IBDV infection in chickens. Keywords: Infectious bursal disease virus, immunosuppression, MAP2K3, IL12B, pro-inflammatory cytokine gene expression

Synergistic effect of zinc oxide-cinnamic acid nanoparticles for wound healing management: in vitro and zebrafish model studies

Wound infections resulting from pathogen infiltration pose a significant challenge in healthcare settings and everyday life. When the skin barrier is compromised due to injuries, surgeries, or chronic conditions, pathogens such as bacteria, fungi, and viruses can enter the body, leading to infections. These infections can range from mild to severe, causing discomfort, delayed healing, and, in some cases, life-threatening complications. Zinc oxide (ZnO) nanoparticles (NPs) have been widely recognized for their antimicrobial and wound healing properties, while cinnamic acid is known for its antioxidant and anti-inflammatory activities. Based on these properties, the combination of ZnO NPs with cinnamic acid (CA) was hypothesized to have enhanced efficacy in addressing wound infections and promoting healing. This study aimed to synthesize and evaluate the potential of ZnO-CN NPs as a multifunctional agent for wound treatment. ZnO-CN NPs were synthesized and characterized using key techniques to confirm their structure and composition. The antioxidant and anti-inflammatory potential of ZnO-CN NPs was evaluated through standard in vitro assays, demonstrating strong free radical scavenging and inhibition of protein denaturation. The antimicrobial activity of the nanoparticles was tested against common wound pathogens, revealing effective inhibition at a minimal concentration. A zebrafish wound healing model was employed to assess both the safety and therapeutic efficacy of the nanoparticles, showing no toxicity at tested concentrations and facilitating faster wound closure. Additionally, pro-inflammatory cytokine gene expression was analyzed to understand the role of ZnO-CN NPs in wound healing mechanisms. In conclusion, ZnO-CN NPs demonstrate potent antioxidant, anti-inflammatory, and antimicrobial properties, making them promising candidates for wound treatment. Given their multifunctional properties and non-toxicity at tested concentrations, ZnO-CN NPs hold significant potential as a therapeutic agent for clinical wound management, warranting further investigation in human models.

Pro-inflammatory cytokines gene expression in liver and kidneys of rats exposed to a sub-lethal dose of Bitis arietans snake venom.

Bitis arietans (Puff adder) is a poisonous snake and its bite causes pain, edema, blistering, tissue damage and neutrophilia. There are limited studies on inflammatory process involved in Bitis arietans envenomation. We therefore investigated the role of proinflammatory cytokines in Bitis arietans venom (BAV)-induced liver and kidney toxicities in rats. Adult male Sprague Dawley rats were treated with BAV (0.5 mg/kg) and were sacrificed after specific time intervals (2 h, 24 h, 1 week). Blood samples were collected for liver and renal function tests and tissues were collected for histopathology and gene expression analysis of IL-1β, IL-6, and TNF-α in liver and kidneys. There was no significant difference in serum ALT activities among different treatment groups. Serum AST was significantly increased at 24 h following BAV injection. In both organs, injection of BAV resulted in mild inflammatory cell infiltration at 2 h post-dosing which normalized after 1 week. In liver, there was a significant increase in IL-1β expression in BAV-treated rats at 2 and 24 h post-dosing that reduced after one week. Significant increases in IL-6 and TNF-α were observed at 24 h and 1 week after BAV exposure. In kidneys, there were significant increases in IL-1β and TNF-α expression at 24 h that subsided after 1 week. In conclusion, a single sub-lethal dose of BAV caused an acute phase inflammation in liver and kidneys. It is most probable that a higher dose of BAV may result in greater and irreversible damage to these organs.