IL-10 Expression Research Articles

IL-23 Promotes γδT Cell Activity in Dry Eye Disease Progression.

Conjunctival-resident γδT cells, the predominant ocular source of interleukin-17A (IL-17A), play crucial roles in dry eye disease (DED) pathogenesis. The upstream regulators of these cells are unknown. This study evaluated the role of conjunctival IL-23 expression in mediating γδT cell generation and elucidated its contribution to dry eye inflammatory responses. Single-cell RNA sequencing (scRNA-seq) was used to identify and quantify conjunctival mRNA molecules in γδT cells in mice. The IL-23 level increased in wild-type (WT) and decreased in γδT-deficient (TCRδ-/-) mice after dry eye was induced via an intelligently controlled environmental system (ICES). Flow cytometry and transcriptome sequencing were used to investigate the impact of the changes in IL-23 expression on human γδT cells. The expression of the IL-23 receptor (IL-23R) was greater in γδT cells than in other conjunctival cell types, such as CD4+ T cells, CD8+ T cells and epithelial cells. An increase in IL-23 led to an increase in γδT cell density, which was proportional to dry eye severity. However, in the TCRδ-/- mice, the upregulation of IL-23 failed to increase the expression level of IL-17A and the severity of dry eye. Furthermore, increases in the expression of IL-23 and the number of γδT cells were evident in the ocular surface cells of patients who developed visual display terminal syndrome. An increase in conjunctival IL-23 expression contributes to the induction of the DED inflammatory response through interactions with its cognate receptor on γδT cells and the promotion of their proliferation. The findings of this study suggest that the suppression of IL-17A through the blockade of IL-23R activation may be a viable target for improving the management of inflammation in DED patients.

Human MAIT cell response profiles biased toward IL-17 or IL-10 are distinct effector states directed by the cytokine milieu

Mucosal-associated invariant T (MAIT) cells are unconventional T cells that mediate rapid antimicrobial immune responses to antigens derived from microbial riboflavin pathway metabolites presented by the evolutionarily conserved MR1 molecules. MAIT cells represent a large pre-expanded T cell subset in humans and are involved in both protective immunity and inflammatory immunopathology. However, what controls the functional heterogeneity of human MAIT cell responses is still largely unclear. Here, combining functional and transcriptomic analyses, we investigate how MAIT cell response programs are influenced by the cytokine milieu at the time of antigen recognition. Activation by MR1-presented antigen together with IL-12 induces intermediate levels of IFNγ and TNF, as well as a regulatory profile with substantial IL-10 production and elevated expression of TIM-3, LAG-3, and PD-1. Activation by the combination of antigen and IL-12 induces a c-MAF-dependent program required for IL-10 production. The MAIT cell-derived IL-10 mediates both autocrine and paracrine immune regulation. In contrast, coactivation of MAIT cells with IL-18 induces IL-17, GM-CSF, IFNγ, and TNF, without IL-10. Notably, IL-18 dominantly counteracts IL-10 expression. The activation states biased toward IL-10 or IL-17 production are reversible and do not represent stable subsets. Finally, MR1-restricted TCR-mediated activation without cytokine coactivation drives primarily granzyme B cytolytic arming. Altogether, these findings demonstrate that human MAIT cells adapt their functional effector response during antigen recognition to cytokine cues in the microenvironment, and identify programs biased toward either regulatory c-MAF-dependent IL-10 expression, or an inflammatory IL-17 and GM-CSF profile.

Lactobacillus rhamnosus GG induces STING-dependent IL-10 in intestinal monocytes and alleviates inflammatory colitis in mice.

Preclinical and clinical observations indicate that the probiotic Lactobacillus rhamnosus GG (LGG) can modulate colonic inflammation. However, the underlying mechanisms have not been explored in depth. Here, we demonstrate that oral administration of live LGG alleviated inflammatory colitis by increasing IL-10 expression in intestinal Ly6C+ monocytes. Mechanistically, LGG induced IL-10 production via the stimulator of IFN genes (STING)/TBK1/NF-κB (RELA) signaling pathway in intestinal Ly6C+ monocytes, enhancing their immune-suppressive function. Elevated IL-10 subsequently activated IL-10 signaling in Ly6C+ monocytes, resulting in an IL-10-based autocrine regulatory loop and inhibition of proinflammatory cytokine production. Furthermore, LGG shifted the gut microbial community and its metabolic functions, leading to intestinal immune responses against colitis. Fecal microbiota transplantation from LGG-colonized mice alleviated immune checkpoint blockade-associated colitis. Our findings highlight the importance of STING signaling in IL-10-dependent antiinflammatory immunity and establish an empirical basis for developing oral administration of live LGG as an efficient and safe therapeutic strategy against inflammatory colitis.

Development of Antigen-Capture Enzyme-linked Immunoassay for Chicken Interleukin-34.

Interleukin-34 (IL-34), a recently identified cytokine, is known to share similar functions with macrophage colony-stimulating factor 1 (CSF-1). However, its function and active regions in avian species have not yet been fully understood. In this study, we report the functional characterization and immunomodulatory properties of chicken IL-34 (chIL-34) using a panel of newly developed anti-chIL-34 mouse monoclonal antibodies (mAbs). Five mAbs (3G11, 10A3, 11A7, 12A8, and 13C2) that specifically recognized recombinant chIL-34 protein were selected and characterized based on their binding activity toward recombinant chIL-34 protein via indirect ELISA and western blotting. A new chIL-34-specific antigen-capture sandwich ELISAs was developed using compatible mAb pairs (3G11 as the capture antibody and biotinylated-12A8 as the detection antibody) which have been identified through pairing assays. To validate the antigen-capture assay, we stimulated native chIL-34 production in the chicken HD11 macrophage cell line using three agonists, Lipopolysaccharides (LPS), polyinosinic:polycytidylic acid (poly I:C), and Resquimod-848 (R-848) at varying concentrations (0.5, 1.0, and 2.0 μg/mL). The qRT-PCR confirmed that the significant expression of chIL-34 was induced by 2.0 μg/mL of LPS, which we selected as an agonist for further chIL-34 production in HD11 cell line. We further stimulated the HD11 cell line with 2.0 μg/mL LPS and monitored the resulting changes in chIL-34 production over time using a mAbs combination (12A8 and biotinylated-3G11). The IL-34 production peaked at 24 hours (h) post stimulation, followed by a decrease at 48 h. This pattern paralleled the expression of chIL-34 mRNA as detected by qRT-PCR. The changes in IL-34 levels in the serum of chickens infected with Eimeria maxima (E. maxima) and Eimeria tenella (E. tenella), measured using a combination of 12A8 and biotinylated-3G11, showed different patterns. Compared to uninfected chickens, IL-34 levels in E. maxima-infected chickens increased starting from 1-day post-infection (dpi), peaked between 3 and 5 dpi, and then rapidly decreased by 7 dpi, falling below the levels observed at 1 dpi. In contrast, IL-34 levels in E. tenella-infected chickens increased from 3 dpi and steadily rose at 5 and 7 dpi. All newly produced mAbs (3G11, 10A3, 11A7, 12A8, and 13C2) specific for chIL-34 effectively neutralized the function of IL-34, as measured by cell proliferation assays and IL-10 expression using qRT-PCR analysis. These new anti-chIL-34 mAbs and the antigen-capture ELISA will be valuable tools for both fundamental and applied research in avian immunology.

Folding expression of IL-32, IL-33 and TNF-α in Patient’s Co-infected with Hepatitis C Virus and Toxoplasmosis

Folding expression of IL-32, IL-33 and TNF-α in Patient’s Co-infected with Hepatitis C Virus and Toxoplasmosis

Activation of Local 11β-Hydroxysteroid Dehydrogenase Type 1 by Diosmetin Enhances Endogenous Glucocorticoid Levels to Alleviate Skin Inflammation: Insights Into a Novel Therapeutic Strategy for Atopic Dermatitis.

Glucocorticoids (GCs) are synthesised de novo by peripheral tissues and the adrenal cortex of the hypothalamic-pituitary-adrenal axis. Skin expresses an enzyme called 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1), which reduces cortisone to the active hormone cortisol which activates GC receptors. 11β-HSD1 plays a significant role in alleviating atopic inflammation through the elevation of the concentrations of active GC in the skin. This study aimed to investigate the role of diosmetin as an activator of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). In human keratinocytes, diosmetin was found to upregulate 11β-HSD1 protein expression and cortisol levels, as well as the transcriptional expression of 11β-HSD1 mRNA. However, this upregulation of 11β-HSD1 mRNA was abrogated in keratinocytes transfected with 11β-HSD1 small interfering RNA (siRNA). In an atopic dermatitis (AD) murine model, topical administration of diosmetin significantly attenuated basal transepidermal water loss (TEWL) and the Eczema Area and Severity Index (EASI), while enhancing stratum corneum (SC) hydration. Diosmetin also increased corticosterone levels in the SC and upregulated 11β-HSD1 expression in both the serum and epidermis. Furthermore, diosmetin treatment led to a marked reduction in serum immunoglobulin E (IgE) and tumour necrosis factor-α (TNF-α) levels, and suppressed mRNA expression of thymic stromal lymphopoietin (TSLP), interleukin-1β (IL-1β), IL-4, and IL-13 in the epidermis. Collectively, these findings suggest that diosmetin promotes the endogenous activation of glucocorticoids via local 11β-HSD1 activation, underscoring its potential as a novel topical therapeutic agent for the management of inflammatory skin disorders, such as AD.

The bitter flavor of Banxia Xiexin decoction activates TAS2R38 to ameliorate low-grade inflammation in the duodenum of mice with functional dyspepsia.

Banxia Xiexin Decoction (BXD) is a traditional herbal formulation with a bitter flavor that has a long-standing history of use in Asia for treating functional dyspepsia (FD). In traditional Chinese medicine, the bitter flavor is believed to play a critical role in the therapeutic activity of BXD. The ethnopharmacological properties of bitter plant extracts are closely associated with their anti-inflammatory effects, which may contribute to their efficacy in FD. However, the specific mechanisms remain unknown. The objective of this study is to uncover the bitter active compounds of BXD and their effects in the treatment of FD. The chemical compounds of BXD were identified using HPLC-Q-Exactive-MS. Active compounds in BXD were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database, and bitter active compounds were further identified using BitterDB and PlantMolecularTasteDB. Molecular docking was employed to identify potential targets of these bitter active compounds, and their activation was validated through flow cytometry analysis of Ca2+. Subsequently, a mouse model of FD was established, and our hypothesis was further validated using enzyme linked immunosorbent assay, immunohistochemistry, immunofluorescence, and western blotting. Through HPLC-Q-Exactive-MS analysis, TCMSP, BitterDB, and PlantMolecularTasteDB database, a total of 11 bitter active compounds in BXD were identified: Baicalein, Baicalin, Berberine, Coptisine, Formononetin, Isorhamnetin, Kaempferol, Naringenin, Palmatine, Quercetin, and Wogonin. Molecular docking results indicated that these active compounds exhibited strong affinity for TAS2R38, with Berberine showing the highest scoring. Flow cytometry analysis of Ca2+ revealed that both Berberine and BXD elevated intracellular calcium concentrations, although this effect was partially antagonized by the TAS2R38 inhibitor probenecid. In vivo experiments demonstrated that BXD effectively improved eosinophil infiltration in the duodenum of FD mice, downregulated the expression of inflammatory factors IL-1β, IL-5, and TNF-α, inhibition of NF-κB signaling pathway activation, alleviated damage to the duodenal mucosal barrier, and reversed gastrointestinal motility disorders, with the therapeutic effect enhancing with increasing doses of BXD. However, this therapeutic effect was partially inhibited following probenecid intervention. BXD contains numerous bitter active compounds that play a significant role in regulating inflammatory activity in the duodenum of FD through the activation of TAS2R38. This finding unveils, for the first time, the ethnopharmacological activity of bitter plant taste agents in anti-inflammatory effects, providing new insights for the treatment and drug development of FD.

REDD1 mediates HDM-induced nuclear-cytoplasmic translocation and release of IL-33 in airway epithelial cells by downregulating Nrf2

ObjectiveThis study aims to investigate whether REDD1 (Regulated in Development and DNA Damage Responses 1) mediates the nuclear-to-cytoplasmic translocation and release of IL-33 in airway epithelial cells induced by house dust mites (HDM).MethodsREDD1 expression levels in bronchial asthma patients were validated using public databases, followed by immunohistochemical analysis of REDD1 protein in airway epithelial cells from these patients. An asthma model was then established using HDM-induced 16HBE cell lines, with REDD1 gene knockout performed. The relationship between varying levels of REDD1 expression, Nrf2, and related inflammatory factors was assessed using Western blot and qPCR. To further investigate the role of the REDD1-Nrf2-IL-33 axis in the development of asthma, we employed Nrf2 activators and inhibitors to reassess the impact of REDD1 on IL-33.ResultsAt both mRNA and protein levels, we found that REDD1 was significantly overexpressed in samples from asthma patients (P < 0.05). In vitro, 24-hour exposure to HDM induced a notable nuclear-to-cytoplasmic translocation of IL-33 and increased its levels in the culture medium of 16HBE cells. In addition, HDM treatment significantly upregulated the expression of both REDD1 and Nrf2. Knockdown of REDD1 markedly suppressed HDM-induced IL-33 release and the expression of TNF-α, IL-6, and IL-1β, while enhancing Nrf2 expression. Moreover, treatment with the Nrf2 agonist curcumin inhibited HDM-induced nuclear-to-cytoplasmic translocation and extracellular secretion of IL-33, whereas the opposite effect was observed when using the Nrf2 antagonist ML385.ConclusionThis study reveals the crucial regulatory role of the REDD1-Nrf2-IL-33 axis in the pathological process of bronchial asthma. REDD1 modulates the expression of IL-33 and other inflammatory factors through the Nrf2 signaling pathway, thereby influencing the onset and progression of asthma.Clinical trial numberNot applicable.

Period circadian clock 3 is crucial for regulation of IL-22-producing type 3 innate lymphoid cells by flavonoids from Shen Ling Bai Zhu San to alleviate colitis.

Type 3 Innate lymphoid cells (ILC3s) functions bear complex response during Inflammatory bowel diseases (IBD). Here, our study first analyzed the main pharmacological components in Shen Ling Bai Zhu San n-butanol extracts (S-Nb), and then explored whether S-Nb administrated immune response of ILC3s, and how it regulates ILC3s. Shen Ling Bai Zhu San (SLBZS) or S-Nb were administrated for 7days to analyze the frequency of ILC3s and their produced cytokine. Using siRNA technology to knock down the expression of period circadian clock 2 (Per2) and period circadian clock 3 (Per3) and Anti-IL-22 antibody was supplied to mice, then detecting the moderator effect of S-Nb on colitis. The most class of S-Nb is flavonoids, with a content of approximately 48%. Oral administration of S-Nb enhanced the production of NCR+ILC3s and IL-22 produced by ILC3s, but did not alter IL-17A. Surprisingly, knocking down the expression of Per3 instead of Per2 inhibited the modulation effect of S-Nb on colitis and reduced IL-22 production, whether originating from NCR+ILC3s or NCR-ILC3s. After neutralizing the expression of IL-22 in mice, S-Nb was deprived of ability to alleviate colitis. The reason why S-Nb alleviates colitis is by enhancing the expression of Per3 via flavonoids, which in turn promotes the secretion of IL-22+ILC3s in intestine.

Circular RNA circ_0061183 regulates microglial polarization induced by airborne ultrafine particles in HMC3 cells via sponging miR-98-5p

Airborne ultrafine particulate matter (PM0.1) can enter the brain, induce microglia activation, and promote the development of Alzheimer’s disease (AD). Circular RNAs (circRNAs) are also involved in AD pathogenesis. However, the role of AD-related circRNAs in PM0.1-induced microglia activation remains unclear. Therefore, we explore cytotoxicity, microglia activation, and AD-associated circRNA expression in human microglial HMC3 cells treated with PM0.1, and further examined circRNA expression in mice and cognitively impaired individuals. The results revealed that PM0.1 exposure decreased cell viability, increased lactate dehydrogenase activity, caused microglia activation, elevated microglial M1 maker expression, downregulated microglial M2 maker expression, and reduced AD-related circ_0061183 expression in vitro. Functionally, circ_0061183 silencing enhanced microglia activation and microglial M1 polarization, but inhibited microglial M2 polarization. Mechanistically, circ_0061183 can bind to miR-98-5p to co-regulate M2 microglial-related IL10 expression, which may affect transforming growth factor-β signaling to regulate PM0.1-inhibited microglial M2 polarization. Moreover, circ_0061183 downregulation was observed in the brain of PM2.5-exposed mice and AD mice and in the blood of cognitively impaired individuals. Furthermore, circ_0061183 was positively related to mini–mental state examination scores and amyloid-β42 peptide expression in elderly individuals. Overall, the current work offers epigenetic insights into the regulatory mechanisms of circRNAs on microglial activation caused by environmental pollutants.

Time of day dependent reduction in stroke infarct volume by the Reverb agonist SR9009 in mice

Ischemic stroke leads to disability and death worldwide and evidence suggests that stroke severity is affected by the time dimension of the stroke. Rev-Erbα regulates the core circadian clock through repression of the positive clock element Bmal1. However, it remains unclear if a Rev-Erbα agonist (SR9009) alleviates stroke pathology in mice. We found that stroke reduces the level of Rev-Erbα and elevates neuroinflammation and stroke severity at zeitgeber time (ZT) ZT06. Therefore, we hypothesized that SR9009 treatment may reduce neuroinflammation and stroke severity in a mouse suture occlusion model. At 12 to 14 weeks, C57BL/6 J (Wild Type, n = 5–10 mice/group) mice were randomly assigned to undergo MCAO stroke for 60 min at either zeitgeber time ZT06 (MCAO-ZT06-sleep phase) or ZT18 (MCAO-ZT18-awake phase). Stroked mice were treated with SR9009 (100 mg/kg) or vehicle at 1 h and 24 h after MCAO. After forty-eight hours of stroke, TTC staining, Western blot, and qRT-PCR were performed. We found that SR9009 treatment alleviates neuroinflammation and infarct volume by Rev-Erb remodeling in ZT06 stroke mice but not in ZT18 stroke mice. Additionally, monocytic and neutrophilic NLRP3 as well as brain NLRP3 levels were reduced by SR9009 treatment in ZT06 stroke though no effects were observed at ZT18 stroke. SR9009 also reduced TNFα expression and increased IL-10 expression in blood and brain in ZT06 stroke mice and no differences were observed at ZT18. There were no significant effects of SR9009 on neurological deficit score and sensorimotor function at ZT06 or ZT18 at 48 h. Our study demonstrates that SR9009 treatment reduces stroke volume, circulating immune response, circadian expression, and that the protection was circadian- and treatment time-dependent.

Interleukin-13 partly induced by the NLRP3 inflammasome promotes Trichinella spiralis encapsulation in infected mice.

Trichinella spiralis infection is a serious parasitic zoonosis in which a collagenous capsule surrounding the larva is developed in the striated muscle cells. However, the mechanism of T. spiralis encapsulation is currently poorly understood. It has been reported that T. spiralis infection can induce the production of IL-13 via the NLRP3 inflammasome, and it has also been suggested IL-13 thus produced may be involved in T. spiralis encapsulation. This research aimed to clarify the involvement of NLRP3 and IL-13 in the T. spiralis capsule formation process. IL-13 and NLRP3 inhibitors were used in a T. spiralis infected mouse model and in C2C12 cells to analyze the role of IL-13 and NLRP3 in encapsulation. The results showed that T. spiralis infection significantly increased the expression levels of IL-13 and collagen IV and VI. The production of collagen around the T. spiralis encapsulation zone was significantly inhibited when an IL-13 inhibitor was applied. Moreover, the expression levels of IL-13 and collagen IV and VI were significantly decreased by the NLRP3 inhibitor in vitro and in vivo. The above results indicated that NLRP3 can participate in the development of T. spiralis encapsulation by regulating IL-13 expression and stimulating collagen IV and VI synthesis during T. spiralis infection.

Combining Single-Cell RNA Sequencing and Mendelian Randomization to Explore Novel Drug Targets for Parkinson's Disease.

Neuroinflammation is a key pathological factor of PD, and T cells play a central role in neuroinflammatory progression. However, the causal effect of T cell-related genes on the risk of PD is still unclear. We explored single-cell RNA sequencing (scRNA-Seq) datasets of the peripheral blood T cells of PD patients and healthy controls, and screened the differentially expressed genes (DEGs) in the cytotoxic CD4 + T cells relative to the other T cell subsets. Pseudo-time series analysis, cell-cell communication analysis, and metabolic pathway analysis was performed for the cytotoxic CD4 + T cells. The DEGs were also functionally annotated through GO and KEGG pathway enrichment analyses. The MR approach was used to establish causal effects of the DEGs (exposure) on PD risk (outcome), and explore new drug targets for PD. The findings of MR analysis were further validated by Steiger filtering, bidirectional MR, Bayesian colocalization analysis, and phenotype scanning, and the GWAS data from an independent PD case-control cohort was used for external validation of the results. Finally, differences in gene expression between PD patients and healthy controls were further validated in scRNA-Seq and bulk transcriptome sequencing data. We found that increased expression of IL-32, GNLY, MT2A, and ARPC2 was significantly associated with a higher risk of PD. In contrast, the increase in ARRB2 was closely related to a lower risk of PD. IL32, GNLY, MT2A, ARRB2, and ARPC2 are the causal genes and potential drug targets of PD. Cytotoxic CD4 + T cells are likely the key effectors of PD-related neuroinflammation. These findings provide new insights into the pathogenesis and treatment options for PD, and further research and clinical trials based on the five potential drug targets and neuroinflammation are necessary.

TRα1mutant suppresses KLF9 to cause endometrial metaplasia with ectopic IL-33 expression leading to uterine fibrosis and infertility.

Thyroid hormone receptors (TRs) mediate the genomic actions of thyroid hormone. Mutations of THRA gene cause a human disease known as resistance to thyroid hormone (RTHα). We created a mouse model expressing a dominant negative mutated TRα1 (Thra1PV/+ mice) that exhibits growth retardation, bone abnormalities, constipation, and anemia, as found in RTHα patients. In addition, female Thra1PV/+ mice exhibit decreased fertility. In the present study, we aimed to characterize the molecular events leading to infertility. Histologically, there was progressive uterine atrophy in Thra1PV/+ mutant mice, characterized by squamous metaplasia of the endometrial mucosa and endometrial fibrosis. RNA-seq analysis of laser-captured micro-dissected endometrium and spatial transcriptomics revealed a key role for Krüppel-like factor (Klf9), a directly-regulated TR target gene, in normal endometrial differentiation. Klf9 was suppressed in the endometrium of mice harboring mutated TRα1 and pathway analysis revealed that deficient Klf9 signaling was associated with squamous differentiation, consistent with the endometrial metaplasia observed histologically. Further, we showed that this metaplastic endometrial mucosa was the source of ectopic IL-33, which was associated with increased T-cell infiltrates, destruction of glands, and endometrial fibrosis. Our studies provide new insights to understand uterine epithelial morphogenesis and how thyroid dysfunction could lead to female infertility.

NLRP3: a key regulator of skin wound healing and macrophage-fibroblast interactions in mice

Wound healing is a highly coordinated process driven by intricate molecular signaling and dynamic interactions between diverse cell types. Nod-like receptor pyrin domain-containing protein 3 (NLRP3) has been implicated in the regulation of inflammation and tissue repair; however, its specific role in skin wound healing remains unclear. This study highlights the pivotal role of NLRP3 in effective skin wound healing, as demonstrated by delayed wound closure and altered cellular and molecular responses in NLRP3-deficient (NLRP3−/−) mice. Histological analysis revealed impaired healing processes, accompanied by reduced expression of key inflammatory mediators, including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2). Deficiencies in apoptosis were evident through altered expression of cysteine-aspartic acid protease 3 (Caspase-3), P53, and B-cell lymphoma-2 (Bcl-2). Furthermore, critical growth factors such as vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and matrix metalloproteinase-9 (MMP-9) were significantly decreased at the excisional skin wound sites. Furthermore, using co-culture systems, we found that NLRP3 mediated the interaction between macrophages and myofibroblasts. Wild-type fibroblast-conditioned media (MFbCM) enhanced nitric oxide (NO), IL-6, and tumor necrosis factor-α (TNF-α) production in M1 macrophages and arginase activity, chitinase 3-like protein 1 (Ym1), and IL-10 expression in M2 macrophages, effects significantly diminished with NLRP3−/− MFbCM. Similarly, conditioned media from wild-type M1 or M2 macrophages promoted the expression of FGF-2, VEGF, and MMP-2 expression in myofibroblasts, which was attenuated when using NLRP3−/− macrophage-conditioned media. PGE2 levels were reduced in both NLRP3−/− macrophages and myofibroblasts. Supplementing NLRP3−/− conditioned media with PGE2 partially restored the impaired functions, suggesting that PGE2 acts as a downstream mediator of NLRP3-regulated macrophage-myofibroblast interactions. These findings indicate that NLRP3 is a key regulator of skin wound healing, facilitating macrophage-myofibroblast communication.

Biogenic silver nanoparticle as an adjuvant in an S1 subunit recombinant vaccine.

Adjuvants are crucial for maintaining specific, protective, and long-lasting immunity. Here, we aimed to evaluate the antigenic and immunogenic activity of a recombinant form of the S1 domain of the Spike protein, associated with biogenic silver nanoparticles (bio-AgNP) and Alhydrogel® as an alternative and conventional adjuvant, respectively, for a SARS-CoV-2 subunit vaccine. We produced and evaluated the antigenicity of the recombinant S1 (rS1) protein by testing its recognition by antibodies present in SARS-CoV-2 positive human serum. The immunogenicity of the rS1 protein was assessed in vivo by its ability to induce antibody production in mice. Furthermore, we sought to establish the types of humoral and cellular immune responses generated through intramuscular immunization in BALB/c mice with the rS1 + bio-AgNP vaccine. The recombinant S1 (rS1) protein was successfully cloned, expressed in Escherichia coli, and confirmed to have strong antigenic potential, being recognized by human antibodies up to a 1:51,200 serum dilution. In vivo assays showed that vaccines using rS1 with either bio-AgNP or Alhydrogel® as adjuvants induced high and consistent antibody titers (1:51,200) and a range of antibody isotypes in mice. Cellular immune response analysis revealed significant IL-10 expression with rS1 + bio-AgNP and both IL-10 and TNFα expression with rS1 + Alhydrogel®. These results support rS1 + bio-AgNP as a promising vaccine candidate for future development, highlighting its potential not only as a viable alternative to traditional adjuvants but also as an innovative contribution to advancing more effective and accessible immunological strategies.

Early embryonic thermal programming and post-hatch flavonoid (Scutellaria baicalensis) supplementation enhanced immune response markers in broiler chickens

IntroductionGenetic selection in broiler chickens has led to increased muscle mass without comparable respiratory and cardiovascular system development, limiting the birds’ capacity to withstand high ambient temperatures and making them vulnerable to heat stress (HS). Early embryonic Thermal Manipulation (TM) has been suggested as an effective and sustainable way to mitigate the adverse effects of HS. This study investigated how these interventions influenced the immune status of broiler chickens exposed to HS.MethodsCobb 500 fertile eggs (n = 600) were incubated according to guidelines. On embryonic day (ED) 12, the eggs were split into two groups: (1) Control, kept at standard temperature until hatch day (ED 21) and (2) Thermal Manipulation (TM), exposed to 38.5°C with 55% humidity for 12 h daily from ED 12 to ED 18. After hatching, chicks were divided into (1) Control, (2) TM, (3) Control under Heat Stress (CHS), (4) TM under Heat Stress (TMHS), (5) Control with Heat Stress and Supplementation (CHSS), and (6) TM with Heat Stress and Supplementation (TMHSS). For the first 21 days, all chicks were raised under normal conditions. From day 22 to day 35, groups CHS, TMHS, CHSS, and TMHSS experienced chronic heat stress (32–33°C for 8 h daily), while the Control and TM groups remained in a thermoneutral environment (22–24°C).Results and discussionTM significantly increased (p &amp;lt; 0.05) AvBD11, IL4, and TLR21 expression in the spleen. TM and baicalein supplementation significantly decreased (p &amp;lt; 0.05) TLR15 expression. In the bursa, TM significantly increased (p &amp;lt; 0.05) IL4 expression. The combination of TM with baicalein significantly increased (p &amp;lt; 0.05) CD3 and decreased (p &amp;lt; 0.05) TLR1 expression. Interestingly, TM alone significantly decreased (p &amp;lt; 0.05) IFNg expression under HS condition. In the thymus, TM significantly decreased (p &amp;lt; 0.05) IL10 and TLR15, while incorporating baicalein with TM decreased (p &amp;lt; 0.05) AvBD6 expression.ConclusionTM improved the immune status of broiler chickens under normal conditions. When combined with baicalein, TM mitigated the negative effects of heat stress by boosting key immune-related gene expression in the spleen, bursa, and thymus.

Hallmarks of Quercetin Benefits as a Functional Supplementary in the Management of Diabetes Mellitus-Related Maladies: From Basic to Clinical Applications.

Quercetin (QE), a particular flavonoid, is well known for its medicinal effects, including anti-oxidant, hypoglycemic, and anti-inflammatory effects. In this review, the findings of QE effects on diabetes STZinduced, alloxan-induced, and its complications have been summarized with a particular focus on in vitro, in vivo, and clinical trials. Consequently, QE mediates several mechanisms, including ameliorating tumor necrosis factor (TNF)-α, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interleukin (IL)-1β, IL-8, and IL-10 expression, increasing insulin glucose uptake to inhibit insulin resistance. Moreover, QE stimulates insulin secretion and attenuates insulin resistance through various pathways, namely transient KATP channel, motivating peroxisome proliferator-activated receptor expression, increasing glucose transporter-4, and decreasing inducible nitric oxide synthase in skeletal muscle. QE has protective effects on the complications caused by diabetes, such as polycystic ovary syndrome, high-fat diet-induced obesity, diabetic-induced hepatic damage, vascular inflammation, nephropathy, and neuropathy.

Saussurea costus alleviates ulcerative colitis by regulating the gut microbiota and improving intestinal barrier integrity

IntroductionThe global health challenge of ulcerative colitis (UC) has been classified by the WHO as a modern refractory disease, commonly referred to as green cancer, with limited treatment options still available, highlighting the urgent need for the development of new therapeutic strategies. Recent pharmacological research has shown that traditional Chinese medicine saussurea costus (SC) possesses beneficial antibacterial and anti-inflammatory properties. Nevertheless, its underlying mechanism remains elusive.MethodsFirstly, we identified the main active components of SC through UHPLC-QTOF-MS analysis. Subsequently, UC mice were induced using DSS and administered different doses of SC to evaluate its efficacy. Additionally, the impact of SC on the repair of the intestinal mucosal barrier was evaluated through immunofluorescence and western blot. Furthermore, 16s rRNA gene sequencing was conducted to elucidate the contribution of gut microbiota to UC pathogenesis.ResultsThe primary components of SC include Proline, Phenylalanine, Isoleucine, Lucidenic acid M, and Pyroglutamic acid. The efficacy of SC was concurrently assessed, revealing its potential to ameliorate histological injury in colitis mice. Furthermore, SC was found to decrease levels of TNF-α, IL-1β, IL-8, and IL-18 while promoting the expression of IL-10 and IL-22. Similarly, we also found that the expression of ZO-1 and Occludin was reversed by SC in colitis mice. In addition, analysis of 16S rRNA gene sequencing indicated that SC reduced harmful bacterial populations, such as Proteobacteria, while simultaneously enhancing the levels of beneficial bacteria like Lactobacillus, thereby contributing to the improvement of UC pathology.ConclusionThis study highlights the therapeutic potential of SC in managing UC through its ability to attenuate inflammatory responses, restore intestinal barrier functionality, and modulate gut microbiota composition, which findings offer insights into potential strategies for advancing UC treatment.

Dietary addition of compound organic acids improves the growth performance, carcass trait, and body health of broilers

IntroductionThe poultry industry constantly seeks strategies to enhance broiler growth performance and overall health. Organic acidifiers, including L-lactic acid, L-malic acid, and acetic acid, have gained attention as potential feed additives to improve animal production by modulating gut health, enhancing nutrient absorption, and supporting immune function. Despite their promising effects in other animal species, the impact of this novel compound organic acidifier on broiler performance, metabolism, and immune response has not been fully elucidated. This study aims to evaluate the effects of this compound acidifier on growth performance, serum lipid profile, antioxidant status, and immune parameters in broilers, providing insights into its potential benefits as a dietary supplement for broiler health and productivity.MethodsA total of 240 broilers were randomly divided into four groups: a control group and three treatment groups receiving 0.25%, 0.5%, or 1.0% acidifier, with six replicates of ten birds each. Over a 6-week period, various parameters were measured, including serum triglycerides, high-density lipoproteins, lysozyme, immunoglobulins (IgA, IgM), superoxide dismutase (SOD) activity, IL-2, TNF-α, and gene expressions related to lipid metabolism.ResultsOver a 6-week period, the acidifier decreased serum triglycerides and high-density lipoproteins while also enhancing growth performance. Additionally, it raised the serum levels of lysozyme, IgA, IgM, and the SOD. Additionally, IL-2 and TNF-α concentrations in the jejunum mucosa decreased. The acidifier upregulated PPARα, AMPK, FABP1 and MTTP expressions, and downregulated APOB100. Overall, the acidifier effectively improved broiler growth performance during the early development phase primarily by enhancing hepatic lipid metabolism, antioxidant capacity, and immune function.ConclusionThese results suggest that the acidifier may accelerate liver lipid metabolism in broilers by modulating the gene expression profiles involved in lipid metabolism.