IL-17 mRNA Expression Research Articles

Association of interleukin 33 gene polymorphisms with susceptibility and regulation of inflammatory mediators in Systemic lupus erythematosus patients.

Studies have indicated the involvement of interleukin (IL)-33 in the pathogenesis of Systemic lupus erythematosus (SLE). This research intended to evaluate the association of IL33 gene rs1929992 and rs7044343 Single nucleotide polymorphisms (SNPs) with risk of SLE. In addition, the association between these SNPs and inflammatory cytokines was determined. In this study, 200 SLE cases and 200 healthy subjects were recruited. Using allelic discrimination Real-time PCR, IL33 gene rs1929992 and rs7044343 SNPs were genotyped. The mRNA expression levels of IL-1β, IL-6, IL-33, TNF-α were determined in the peripheral blood mononuclear cells (PBMCs). The serum levels of cytokines were also measured. The G allele (OR = 1.57, CI: 1.18-2.08, P = 0.0017), GG genotype (OR = 2.52, CI: 1.33-4.77, P = 0.0043), and GA genotype (OR = 2.12, CI: 1.34-3.34, P = 0.0011) of rs1929992 SNP was significantly associated with an increased SLE risk. The C allele (OR = 1.44, CI: 1.08-1.90; P = 0.0105), CC genotype (OR = 2.07, CI: 1.15-3.71; P = 0.0146), and CT genotype (OR = 1.61, CI: 1.02-2.53, P = 0.0395) of rs7044343 was significantly associated with increased SLE risk. The PBMC mRNA expression and serum levels of IL-1β, IL-6, IL-33, TNF-α were significantly increased in the SLE patients compared to controls. However, there was no significant difference in the mRNA expression and serum levels of IL-1β, IL-6, IL-33, and TNF-α among the SLE patients with three genotypes for both rs1929992 and rs7044343 polymorphisms. IL33 gene rs1929992 and rs7044343 SNPs are involved in SLE pathogenesis but they might not influence on the inflammatory pathway.

Immune phenotype of the endometrium in patients with recurrent implantation failures after the transfer of genetically tested embryos in assisted reproductive technology programs

Recurrent implantation failures (RIF) in assisted reproduction programs are one of the most challenging problems. Among the factors that can adversely affect implantation, endometrial immune structural disorders may be one of the leading causes. The aim of our work was to study the immune features of the endometrium in women with RIF after genetically tested embryo transfer in comparison with fertile gestational carriers. Immune cells in endometrial samples were studied by flow cytometry and RNA expression of IL (interleukin)15, IL18, fibroblast growth factor-inducible 14 receptor (Fn14), and tumor necrosis factor-like weak inducer of apoptosis (TWEAK) by reverse polymerase chain reaction. In one-third of the cases, a unique immune profile of the endometrium, which we called the not transformed endometrial immune phenotype, was found. It is characterized by a combination of features, such as high expression of HLA-DR on natural killers (NK), increased fraction of CD16 + , and a decreased fraction of CD56bright endometrial NK. In addition, when compared to gestational carriers, patients with RIF had a greater discrepancy between IL18 mRNA expression data, reduced mean TWEAK and Fn14 levels, and increased IL18/TWEAK and IL15/Fn14 ratios. Immune abnormalities that were found in more than half of the patients (66.7 %) may be the cause of implantation failures in genetically tested embryo transfer programs.

Fermented Wheat Bran Polysaccharides Improved Intestinal Health of Zebrafish in Terms of Intestinal Motility and Barrier Function

Intestinal barrier dysfunction and gut microbiota disorders have been associated with various intestinal and extraintestinal diseases. Fermented wheat bran polysaccharides (FWBP) are promising natural products for enhancing the growth performance and antioxidant function of zebrafish. The present study was conducted, in order to investigate the effects of FWBP on the intestinal motility and barrier function of zebrafish, which could provide evidence for the further potential of using FWBP as a functional food ingredient in the consideration of gut health. In Experiment 1, the normal or loperamide hydrochloride-induced constipation zebrafish larvae were treated with three concentrations of FWBP (10, 20, 40 μg/mL). In Experiment 2, 180 one month-old healthy zebrafish were randomly divided into three groups (six replicates/group and 10 zebrafish/tank) and fed with a basal diet, 0.05% FWBP, or 0.10% FWBP for eight weeks. The results showed that FWBP treatment for 6 h can reduce the fluorescence intensity and alleviate constipation, thereby promoting the gastrointestinal motility of zebrafish. When compared with control group, zebrafish fed diets containing FWBP showed an increased villus height (p < 0.05), an up-regulated mRNA expression of the tight junction protein 1α, muc2.1, muc5.1, matrix metalloproteinases 9 and defensin1 (p < 0.05), an increased abundance of the phylum Firmicutes (p < 0.05), and a decreased abundance of the phylum Proteobacteria, family Aeromonadaceae, and genus Aeromonas (p < 0.05). In addition, 0.05% FWBP supplementation up-regulated the intestinal mRNA expression of IL-10 and Occludin1 (p < 0.05), enhanced the Shannon and Chao1 indexes (p < 0.05), and increased the abundance of Bacteroidota and Actinobacteriota at the phylum level (p < 0.05). Additionally, 0.1% FWBP supplementation significantly improved the villus height to crypt depth ratio (p < 0.05) and increased the mRNA expression of IL-17 (p < 0.05). These findings reveal that FWBP can promote the intestinal motility and enhance the intestinal barrier function, thus improving the intestinal health of zebrafish.

Dietary glutamate enhances intestinal immunity by modulating microbiota and Th17/Treg balance-related immune signaling in piglets after lipopolysaccharide challenge

The purpose of this study was to explore the effects of glutamate on piglet growth performance and intestinal immunity function, and to further elucidate its mechanism. In a 2 × 2 factorial design involving immunological challenge (lipopolysaccharide (LPS) or saline) and diet (with or without glutamate), twenty-four piglets were randomly assigned to four groups, each with 6 replicates. Piglets were fed with a basal or glutamate diet for 21 d before being injected intraperitoneally with LPS or saline. Piglet's intestinal samples were collected 4 h after injection. Results showed that glutamate increased daily feed intake, average daily gain, villus length, villus area, and villus length to crypt depth ratio (V/C), and decreased the crypt depth (P < 0.05). Furthermore, glutamate increased the mRNA expression of forkhead box P3 (FOXP3), a signal transducer and activator of transcription 5 (STAT5) and transforming growth factor beta, while decreasing the mRNA expression of RAR-related orphan receptor c and STAT3. Glutamate increased interleukin-10 (IL-10) mRNA expression while decreasing the mRNA expression of IL-1β, IL-6, IL-8, IL-17, IL-21, and tumor necrosis factor-α. At the phylum level, glutamate increased the Actinobacteriota abundance and Firmicutes-to-Bacteroidetes ratio while decreasing Firmicutes abundance. At the genus level, glutamate improved the abundance of beneficial bacteria (e.g., Lactobacillus, Prevotellaceae-NK3B31-group, and UCG-005). Furthermore, glutamate increased the concentrations of short-chain fatty acids (SCFAs). Correlation analysis revealed that the intestinal microbiota is closely related to Th17/Treg balance-related index and SCFAs. Collectively, glutamate can improve piglet growth performance and intestinal immunity by modulating gut microbiota and Th17/Treg balance-related signaling pathways.

The effect of lipid-lowering therapies on the pro-inflammatory and anti-inflammatory properties of vascular endothelial cells.

Atherosclerosis and cardiovascular events can be prevented, or treated, using statin therapy, either alone or in combination with ezetimibe. Chronic inflammation, vascular proliferation, and the development of atherosclerosis are also influenced by 25-hydroxycholesterol (25-OHC). The aim of the study was to compare the direct pleiotropic effects of two commonly-used statins (atorvastatin, rosuvastatin), ezetimibe, and their combinations, on the mRNA expression of pro-inflammatory IL1β, IL-18 and IL-23 and anti-inflammatory TGFβ, IL-35 (EBI3, IL-12 subunits), IL-10 and IL-37, in endothelial cells damaged by 25-OHC. It also analyzed IL-35 expression at the protein level. HUVECs were stimulated with atorvastatin (5 μM), rosuvastatin (10 μM), ezetimibe (1.22 μM), atorvastatin-ezetimibe (5 μM + 1.22 μM) or rosuvastatin-ezetimibe (10 μM + 1.22 μM), with or without pre-incubation with 10 μg/mL 25-OHC. mRNA expression was analyzed by real-time PCR. The protein level of IL-35 was analyzed by ELISA. In the pre-stimulated HUVECs, atorvastatin and rosuvastatin decreased mRNA expression of IL1β, IL-18, IL-23, TGFβ, IL35 and increased mRNA expression of IL-10 and IL-37 compared to 25-OHC. Furthermore, only incubation with rosuvastatin and rosuvastatin-ezetimibe decreased IL-35 mRNA and protein levels. Ezetimibe down-regulated only IL1β. Treatment with rosuvastatin-ezetimibe and atorvastatin-ezetimibe reversed the effect of 25-OHC in IL1β, IL-18 and IL-35 mRNA expression. In conclusion, rosuvastatin has the strongest anti-inflammatory effects and is the best at reducing the effect of oxysterols. Both statins exert a greater anti-inflammatory effect than ezetimibe. The anti-inflammatory effect of the combination therapies appears to be based on the effects of the statins alone and not their combination with ezetimibe.

P094 Impacts of 5-aminosalicylic acid on the intestinal microbiota leading to the anti-inflammatory effect

Abstract Background 5-aminosalicylic acid (5-ASA) is widely used for the treatment of inflammatory bowel disease (IBD), while its anti-inflammatory mechanisms remain controversial. The intestinal microbiota plays a certain role in the pathogenesis of IBD. We hypothesised that 5-ASA exerts anti-inflammatory effects by altering the gut microbiota. Elucidating the effects of 5-ASA on gut microbiota and its contribution to anti-inflammatory effects can lead to the development of effective and economical IBD medications. Methods Specific-pathogen-free (SPF) mice were orally administered 5-ASA for 4 weeks. The fecal bacterial compositions were examined with 16S rRNA gene amplicon sequencing analysis in mice of the 5-ASA-treatment group (5-ASA group) and no-treatment group (NT group). The immune profiles of the colonic mucosa were assessed. The feces from female mice in both groups were gavaged into germ-free (GF) female mice in separated isolators. In each isolator, these female mice (5-ASA dams and NT dams) were mated with GF male mice. The populations of CD4+ T cells in mesenteric lymph nodes (MLN) and colonic mucosal immune profiles were analysed in their pups. The pups were used for the dextran sulfate sodium (DSS)-induced colitis model to evaluate the anti-inflammatory effects of the gut microbiota altered by 5-ASA. Results The fecal bacterial compositions changed with 5-ASA treatment over time in SPF mice. At the genus level, the relative abundance of Allobaculum increased significantly in the females of the 5-ASA group compared to the NT group (corrected p &amp;lt; 0.05). The same tendency was observed in male counterparts. The mRNA expression of IL-10 and IL-22 in the colonic mucosa increased significantly in mice of the 5-ASA group compared to their counterparts (p &amp;lt; 0.05 in both sexes). In each isolator, the fecal bacterial compositions were similar between the dams and pups, suggesting that the gut microbiota was vertically transmitted from the dams to the pups in each group. CD4+RORgt+ T cells increased in MLN in the pups of 5-ASA dams compared to those of NT dams (p &amp;lt; 0.01). The mRNA expression of TGF-b, claudin-2 and claudin-3 in the colonic mucosa increased significantly (p &amp;lt; 0.001). The colonic inflammation induced by DSS was milder in 5-ASA pups compared to NT pups (Figure 1). Conclusion Orally-administered 5-ASA altered the intestinal microbiota affecting the host immune system and leading to less susceptibility to DSS-induced colitis. Alteration of gut microbiota could be a part of the anti-inflammatory mechanisms of 5-ASA.

Relationship between IL-22 and IL-22BP in diabetic cognitive dysfunction.

CD4 + T helper (Th)22 cells play a regulatory role in autoimmune diseases such as type 1 diabetes mellitus. The Th22-related cytokine interleukin (IL)-22, the expression of which is increased in diabetes mellitus (DM), can act as a neurotrophic factor to protect neurons from apoptosis. Paradoxically, neuronal apoptosis and learning and memory decline occur in DM. In this study, we investigated the relationship between IL-22 and its receptors IL-22Rα1 and IL-22 binding protein (IL-22BP, a soluble inhibitor of IL-22) in diabetic encephalopathy (DE) and the effects of IL-22 on hippocampal neurons, learning and memory. A C57BL/6 mouse model of diabetes was constructed by intraperitoneal injection of streptozotocin. The mice were randomly divided into 4 groups: the control group, diabetes group, diabetes + recombinantIL-22 (rIL-22) group and diabetes + IL-22BP group. The Morris water maze test was used to evaluate learning and memory, the expression of IL-22 was measured by ELISA, and Evans Blue staining was used to evaluate blood-brain barrier permeability. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to measure the mRNA expression of IL-22 and IL-22Rα1 in the hippocampus. The morphology and number of hippocampal neurons were assessed by Nissl staining, and TUNEL staining was used to detect hippocampal neuronal apoptosis. Immunofluorescence was used to analyze IL-22Rα1 expression and localization in hippocampus, and Western blotting was used to evaluate the expression of IL-22, IL-22Rα1, IL-22BP, and the apoptosis related proteins Caspase-3 and C-caspase-3. Compared with those in the control group, mice in the diabetes group showed cognitive decline; apoptosis of hippocampal neurons; increased expression of hippocampal Caspase-3, C-Caspase-3, IL-22, IL-22Rα1, and IL-22BP; and a decreased IL-22/IL-22BP ratio. Learning and memory were improved, neuronal apoptosis was attenuated, IL-22Rα1 expression and the IL-22/IL-22BP ratio were increased, and caspase-3 and C-caspase-3 expression was decreased in the rIL-22-treated group compared with the diabetes group. IL-22BP treatment aggravated diabetic cognitive dysfunction and pathological alterations in the hippocampus, decreased the IL-22/IL-22BP ratio, and increased the expression of caspase-3 and C-caspase-3 in mice with diabetes. A decrease in the IL-22/IL-22BP ratio plays an important role in diabetic cognitive dysfunction, and rIL-22 can effectively alleviate DE. Herein, we shed light on the interaction between IL-22 and IL-22BP as therapeutic targets for DM.

The prognostic value and immune correlation of IL18 expression and promoter methylation in renal cell carcinoma

BackgroundRenal cell carcinoma (RCC) is not sensitive to immunotherapy and has poor prognosis. DNA methylation regulates gene expression, and its abnormal changes are related to many human diseases. Recently, DNA methylation has been found to participate in immune infiltration in various cancers. However, its pattern in RCC remains poorly understood.ResultsWe found that IL18 was significantly over-expressed in RCC tumor tissues compared to normal adjacent tissues The IL18 promoter region was hypomethylated, which was strongly correlated with elevated IL18 mRNA expression, and predicted advanced clinicopathological characteristics and shorter overall survival. Furthermore, we found that IL18 promoter methylation was significantly related to the down-regulation of immune checkpoint molecules and increase of CD8 + T cell infiltration in RCC tumor tissues.ConclusionsWe have identified the important role of IL18 promoter methylation and expression, which are associated with clinicopathological characteristics, overall survival, immune cell infiltration and expression of immune checkpoint molecules in RCC. We present the rationale for IL18 promoter methylation as a molecular biomarker for predicting the response of RCC to immune checkpoint inhibitors.

Protective effects of Fagopyrum dibotrys on oxidized oil-induced oxidative stress, intestinal barrier impairment, and altered cecal microbiota in broiler chickens

The objective of this study was to evaluate protective effects of Fagopyrum dibotrys on antioxidant ability, intestinal barrier functions, and cecal microbiota in broiler chickens fed oxidized soybean oil. A total of 640 male Tiejiaoma broilers were randomly assigned to 8 treatments with 8 cages (10 birds per cage), as follows: birds fed basal diets containing fresh soybean oil and 0, 0.5, 1, or 2% F. dibotrys (FSCON, FSFAL, FSFAM, and FSFAH, respectively), and birds fed basal diets containing oxidized oil and 0, 0.5, 1, or 2% F. dibotrys (OSCON, OSFAL, OSFAM, and OSFAH). Oxidized oil significantly decreased transcription of Nrf2 and its downstream genes, including CAT and SOD1 in the jejunal mucosa, increased jejunal mucosa IL-6 mRNA expression, and decreased jejunal mucosa IL-22 mRNA expression and downregulated Claudin-1 and ZO-1; however, all these effects were reversed by F. dibotrys. Either 1 or 2% F. dibotrys alleviated the decreased liver SOD induced by oxidized oil on d 42. The decreased SOD and GPX, and increased MDA induced by oxidized oil were reversed by adding 1 or 2% F. dibotrys in jejunal mucosa. In addition, based on 16S rDNA, 2% F. dibotrys promoted the Firmicutes phylum and Candidatus_Arthromitus genera, but suppressed the Proteobacteria phylum and Streptococcus, Enterococcus, and Escherichia genera. In summary, oxidative stress induced by oxidized oil was ameliorated by F. dibotrys upregulating transcription of Nrf2 and its downstream genes to restore redox balance, reinforcing the intestinal barrier via higher expression of Claudin-1/ZO-1, ameliorating the inflammatory response by regulating expression of IL-6 and IL-22, and facilitating growth of Candidatus_arthromitus in the cecum. Therefore, F. dibotrys has potential as a feed additive for poultry by ameliorating oxidative stress caused by oxidized oil, enhancing barrier function, and improving gut microbiome composition.

CCL20/CCR6 chemokine signaling is not essential for pathogenesis in an experimental autoimmune encephalomyelitis mouse model of multiple sclerosis

Multiple sclerosis is an autoimmune disease in which the immune system attacks the nerve myelin sheath. The balance between pathogenic Th17 cells and regulatory Treg cells, both of which express the chemokine receptor CCR6 is critical for determining disease activity. It has been postulated that CCL20, the cognate ligand of CCR6, produced by the blood-brain barrier attracts these immune cells to the central nervous system (CNS). However, the pathological phenotypes of the experimental model of multiple sclerosis in CCR6-knockout (KO) mice are inconclusive, while this has not been addressed in CCL20-KO mice.To address this, we generated CCL20-KO and CCR6-KO mice using the CRISPR/Cas9 system. Clinical phenotypes of experimental autoimmune encephalomyelitis (EAE) in the chronic phase were slightly exacerbated in both mutant mice relative to those in wild-type (WT) mice. Inflammatory cell infiltration and demyelination in the CNS were similar in the KO and WT mice. CNS CD4+ T cell counts were the same for mutant and WT mice. The mutant and WT mice did not differ significantly in the proportions of Th17 and Treg cells in the CNS, or in IL-17 and TGF-β mRNA expression in the CNS.These findings suggest that CCL20/CCR6-mediated cell migration is not necessarily required for the onset of EAE, and may be compensated for by other chemokine signals.

A novel gut-restricted RIPK1 inhibitor, SZ-15, ameliorates DSS-induced ulcerative colitis

As a key mediator of cell death and inflammation, receptor-interacting protein kinase 1 (RIPK1) responds to a broad set of inflammatory and pro-death stimuli in human diseases. Inhibitors targeting RIPK1 are being investigated for the treatment of a wide range of human diseases, including ulcerative colitis. In the present study, we designed, synthesized, and investigated the anti-necroptosis and RIPK1-inhibition effects of SZ-15—a symmetrical high-molecular-weight (>500 Da) compound. SZ-15 effectively inhibited necroptosis in U937 and HT-29 cells at concentrations of 1 nM and 10 nM, respectively, and SZ-15 at a concentration of 10 nM almost completely blocked RIPK1, RIPK3, and mixed-lineage kinase domain-like (MLKL) protein phosphorylation induced by necrosis inducers. SZ-15 suppressed the pro-necroptosis function of RIPK1 by downregulating the mRNA expression of pro-inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6. The activities of SZ-15 were effectively restricted to the gut: The percent recovery of the parent form of SZ-15 in mouse feces was 85.75%. Nevertheless, SZ-15 was effectively absorbed and detected in colon tissues after 1 h at a concentration of 3335 ± 868 ng/g, indicating that membrane permeability was maintained. SZ-15 alleviated dextran sulfate sodium (DSS)-induced ulcerative colitis in vivo by decreasing TNF-α, IL-1β, IL-22, and IL-6 mRNA expression in colonic tissues. Our preclinical study describes a novel gut-restricted RIPK1 inhibitor that shows great potential for use in the clinical treatment of ulcerative colitis.

Assessing Antipsoriatic Effects of Bitter Pu’er Tea and Its Three Major Compounds, Strictinin, Theacrine and Epigallocatechin Gallate, in Imiquimod-Treated Mice

Psoriasis is a chronic inflammatory skin disease with hyperproliferation and aberrant differentiation of keratinocytes in association with the elevation of interleukin-17A (IL-17A) and IL-23 levels. In an animal model, psoriasis-like dermatitis was induced on the shaved dorsal skin of BALB/c mice by topical application of imiquimod (IMQ), a synthetic ligand of Toll-like receptor 7. Administration of bitter Pu’er tea significantly reduced psoriasis-like dermatitis in IMQ-treated mice, including a reduction in dorsal skin lesions, splenomegaly and the mRNA expression levels of IL-17A and IL-23. To examine putative antipsoriatic constituents, three major compounds in bitter Pu’er tea, strictinin, theacrine and epigallocatechin gallate (EGCG), were separately given as supplements to IMQ-treated mice. The results showed that all the three compounds attenuated the severity of psoriasis by reducing epidermal thickness. Only theacrine significantly attenuated splenomegaly. All the three compounds inhibited the expression of IL-23 mRNA in the skin as well as reduced the content of IL-17A+CD4+ T cells in the spleen, and strictinin was found to be relatively effective. It seemed that the antipsoriatic activity of bitter Pu’er tea was attributed to the additive effects of its multiple active compounds.

Interleukin 23 receptor gene polymorphisms and their role in the inflammatory status of rheumatoid arthritis patients in an Iranian population.

Several investigations have disclosed the involvement of the interleukin (IL)-23/IL-17 pathway in rheumatoid arthritis (RA) pathogenesis. Here we investigated the association of single nucleotide polymorphisms (SNPs) in the IL23 receptor (IL23R) gene with RA risk. In addition, the role of these SNPs with the inflammatory state of the patients were determined. In this case-control study, 200 RA cases and 200 healthy subjects were recruited. Using allelic discrimination real-time polymerase chain reaction, both IL23R rs10489629 and rs1004819 SNPs were genotyped. The messenger RNA (mRNA) expression levels of IL-23R, IL-23, and IL-17A were determined in peripheral blood mononuclear cells (PBMCs). The serum levels of IL-23 and IL-17A were also determined. The A allele (odds ratio [OR]=1.52, 95% CI: 1.15-2.01; P= .0030), AA genotype (OR=2.41, 95% CI: 1.33-4.35; P= .0035), and AG genotype (OR=2.55, 95% CI: 1.56-4.16, P= .0002) of rs1004819 SNP was significantly associated with increased RA risk. The mRNA expression of IL-17A (fold change=2.55, P= .00027), IL-23 (fold change=1.62, P= .0081), and IL-23R (fold change=1.59, P= .0077) was significantly upregulated in the PBMCs from RA patients compared to that of healthy controls. Serum levels of IL-17A (P= .00019) and IL-23 (P= .00055) was significantly higher in the RA patients compared to the controls. No significant association was detected between patient data and SNPs. The IL-23/IL-27 pathway plays a role in RA pathogenesis, but IL23R gene rs1004819 SNP might not be regulating this pathway in RA disease.

Gastric bypass surgery weight loss-independently induces gut Il-22 release in association with improved glycemic control in obese Zucker fatty rats

BackgroundRoux-en-Y gastric bypass surgery (RYGB) improves glycemic control in individuals with severe obesity beyond the effects of weight loss alone. To identify potential underlying mechanisms, we asked how equivalent weight loss from RYGB and from chronic caloric restriction impact gut release of the metabolically beneficial cytokine interleukin-22 (Il-22). MethodsObese male Zucker fatty rats were randomized into sham-operated (Sham), RYGB, and sham-operated, body weight-matched to RYGB (BWM) groups. Food intake and body weight were measured regularly for 4 weeks. An oral glucose tolerance test (OGTT) was performed on postoperative day 27. Portal vein plasma, systemic plasma, and whole-wall samples from throughout the gut were collected on postoperative day 28. Gut Il-22 mRNA expression was determined by real-time quantitative PCR. Plasma Il-22 levels were determined by enzyme-linked immunosorbant assay (ELISA). ResultsRYGB and BWM rats had lower food intake and body weight as well as superior blood glucose clearing capability compared with Sham rats. RYGB rats also had superior blood glucose clearing capability compared with BWM rats despite having similar body weights and higher food intake. Il-22 mRNA expression was approximately 100-fold higher specifically in the upper jejunum in RYGB rats compared with Sham rats. Il-22 protein was only detectable in portal vein (34.1 ± 9.4 pg/mL) and systemic (46.9 ± 10.5 pg/mL) plasma in RYGB rats. Area under the curve of blood glucose during the OGTT, but not food intake or body weight, negatively correlated with portal vein and systemic plasma Il-22 levels in RYGB rats. ConclusionsThese results suggest that induction of gut Il-22 release might partly account for the weight loss-independent improvements in glycemic control after RYGB, and further support the use of this cytokine for the treatment of metabolic disease.

Effects of lutein on jejunal mucosal barrier function and inflammatory responses in lipopolysaccharide-challenged yellow-feather broilers

Broilers are frequently exposed to various immunological stresses, which lead to intestinal damage, weakened immunity, and even growth retardation. Lutein, as a kind of carotenoid, possesses antioxidant and immunomodulatory functions. Therefore, this study was conducted to investigate the effects of lutein on jejunal mucosal barrier function and inflammatory responses of yellow-feather broilers challenged with lipopolysaccharide (LPS). A total of two hundred eight-eight 1-day-old yellow-feather broilers were randomly allocated to 3 groups with 8 replicate cages containing 12 birds each. Birds were fed broken-rice-soybean basal diet containing 0, 20 and 40 mg/kg lutein (CON, LU20 and LU40) for 26 d. On days 21, 23, and 25 of the trial, broilers were intraperitoneally injected with LPS (1 mg/kg body weight). The results showed that, compared with CON group, LU40 supplementations significantly increased the average daily gain (ADG) of broilers at 1 to 21 and 22 to 26 d of age (P < 0.05), significantly decreased the ratio of feed to gain (F/G) of broilers at 22 to 26 d of age (P < 0.05). LU20 and LU40 supplementations increased goblet cell density in jejunum of broilers under LPS challenge, and LU20 supplementation elevated the villus area (P < 0.05). Scanning electron microscopy of jejunal mucosa revealed significant villi damage, while transmission electron microscopy demonstrated severe enterocyte damage and loss of cellular integrity in CON group. In particular, mitochondria were morphologically altered, appearing irregular or swollen. Apical junctional complexes between adjacent enterocytes were obviously shorter and saccular in CON group. LU20 and LU40 supplementations increased the mRNA expressions of Occludin, Claudin-1, and ZO-1 in the jejunal mucosa of broilers under LPS challenge (P < 0.05), restrained TLR4/MyD88/NF-κB pathway activation in the jejunal mucosa, decreased the mRNA expressions of IL-1β and IL-6, and strengthened the mRNA expressions of IL-4 and IL-10 (P < 0.05). Meanwhile, the protein expressions of p38 and JNK in LU40 group were lower than CON group (P < 0.05). It can be concluded that 40 mg/kg lutein supplementation improved LPS-induced jejunal mucosal barrier function and tamed inflammation of yellow-feather broilers.

Effect of Qing Chang oral liquid on the treatment of artificially infected chicken coccidiosis and the cellular immunity.

Qing Chang oral liquid (QOL) is a veterinary drug, which mainly composed of Artemisiae annuae herba, Dichroae radix, Agrimonia pilosa and Sanguisorbae radix. This study aims to explore the effect of Qing Chang Oral Liquid (QOL) on the treatment effect of artificially infected chicken coccidiosis and to the cellular immunity. Healthy Roman chickens were randomly divided into five groups: blank group, model group, QOL high-, medium- and low-dose groups. All the groups were orally administered with 1 × 104 sporulated oocysts (except the blank group). After 5 days of oral administration, the high-, medium- and low-dose groups of QOL were added to the drinking water at 2.4, 1.8 and 1.2ml/kg, respectively. The blank and model groups were fed normally, and this experiment lasted for 7 days. The clinical signs were observed, and the relative weight gain, survival rate, cecum lesion score and oocyst value were measured to evaluate the effect of QOL. Meanwhile, the peripheral blood T-lymphocyte subsets and cecal IL-2, IL-17, IFN-γ mRNA expression were detected by flow cytometry and fluorescent quantitative PCR. The chickens in the model group were in poor mental state, gathered together, had loose stools or bloody stools and had less food intake and less exercise. The chicken mental state improved, the food intake and drinking water increased, and the faeces are normal in the high-, medium- and low-dose groups, especially in the high-dose group, the anti-coccidial indexes were up to 173.08. No significant differences were observed (p > 0.05) in the peripheral blood CD3+ , CD3+ CD4+ between the experimental groups. Compared with the blank group, there were different degrees of increase in each dose drug group of the cecal IFN-γ, IL-2 and IL-17 mRNA expression, but the high-dose group was significantly reduced compared with the model group (p < 0.01), but there was no significant difference (p > 0.05). This study suggests that QOL has positive anti-coccidial effect but has no obvious effect on the cellular immunity.

PI3K/AKT/mTOR Signaling Pathway Is Downregulated by Runzaoling (RZL) in Sjögren's Syndrome.

Infiltration and aggregation of lymphocytes in exocrine glands are the basic pathological manifestations of Sjögren's syndrome (SS), and the incidence of SS has been increasing year by year in recent years. To explore the potential signaling pathway of Runzaoling (RZL) in alleviating SS, the possible targets of RZL in SS were firstly explored through network pharmacology, and then, the regulation of PI3K/AKT/mTOR signaling in NOD mice and Th17 cells was verified. 75 8-week-old NOD mice were casually classified into 5 groups: model; hydroxychloroquine; high, medium, and low dose RZL groups, with 15 in each; and 15 BALB/c mice were employed as control group. After 10 weeks of continuous intragastric administration in mice and 24 hours of drugs intervention in Th17 cells, histopathology was observed by HE staining, and the gene transcription levels were identified by real-time quantitative PCR (RT-qPCR). The protein expressions were detected by western blotting (WB). The findings showed that high and medium dose RZL group could attenuate the submandibular gland tissue damage. The results indicated that the mRNA expressions of PI3K, AKT, mTOR, STAT3, and IL-17 in SS mice and in IL-17 stimulation of Th17 cells were dramatically increased compared with control group and decreased to varying degrees after RZL intervention. The trend of phosphorylated PI3K/AKT/mTOR and STAT3 and IL-17 protein expression in NOD mice and Th17 cells were consistent with mRNA. RZL can downregulate STAT3 and IL-17 expressions in the submandibular gland of NOD mice and in Th17 cells via regulating the PI3K/AKT/mTOR signaling pathway. Moreover, RZL could reduce the activation of CD4+ T lymphocyte differentiation to Th17 cells.

Effect of dietary resveratrol supplementation on growth performance, antioxidant capacity, intestinal immunity and gut microbiota in yellow-feathered broilers challenged with lipopolysaccharide

Resveratrol (RES) displays strong antioxidant and anti-inflammatory properties in protecting the animals from various stressors and inflammatory injuries, but its interrelationship with the gut microbiota remained largely unclear. This study was carried out to investigate the effects of dietary RES supplementation on growth performance, antioxidant capacity, intestinal immunity and gut microbiota in yellow-feathered broilers challenged by lipopolysaccharide (LPS). A total of 240 yellow-feathered broilers were randomly assigned to four treatment groups in a 2 × 2 factorial design. The broilers were fed with the control diet or control diet supplemented with 400 mg/kg RES, followed by challenge with LPS or the same amount of saline. Dietary RES supplementation significantly alleviated the decreases in the final body weight (BW), average daily gain (ADG), and ADFI induced by LPS (P < 0.05). LPS challenge significantly increased plasma concentrations of triglyceride, high-density lipoprotein cholesterol (HDL-C), aspartate aminotransferase (AST), and cortisol levels, but decreased triiodothyronine (T3) and insulin levels (P < 0.05). Dietary supplementation with RES significantly reversed the elevated creatinine concentrations and the decreased concentrations of T3 and insulin caused by LPS (P < 0.05). Moreover, dietary RES supplementation significantly increased plasma total antioxidant capacity (T-AOC) and catalase (CAT) activities and superoxide dismutase (SOD) and T-AOC activities in jejunal mucosa and reduced malondialdehyde (MDA) concentration in the plasma (P < 0.05). The reduction in the villus height to crypt depth ratio in duodenum, jejunum and ileum and the shortening of villus height in jejunum and ileum caused by LPS were also alleviated by RES treatment (P < 0.05). Furthermore, the increased concentrations of intestinal tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β caused by LPS were significantly decreased by RES treatment (P < 0.05). Dietary RES treatment increased the mRNA expression of claudin-1, claudin-5, occludin, and zonula occludens-1 (ZO-1), and decreased mRNA expression of IL-1β, IL-8, IL-17, and TNF-α after LPS challenge (P < 0.05). Dietary RES treatments significantly decreased the dominance of cecal microbiota, and increased the Pieiou-e and Simpson index. Moreover, dietary RES supplementation increased relative abundance of UCG_ 009, Erysipelotrichaceae, Christensenellaceae_R-7_group, Anaerotruncus, RF39, and Ruminococcus while decreasing the abundance of Alistipes at genus level. Spearman correlation analysis revealed that the microbes at the order and genus levels significantly correlated with indicators of growth performance, antioxidant capacity, and intestinal health. Collectively, dietary supplementation with 400 mg/kg RES could improve growth performance and antioxidant capacity, and modulate intestinal immunity in yellow-feathered broilers challenged by LPS at early stage, which might be closely associated with the regulation of gut microbiota community composition.

Anemoside B4 ameliorates experimental autoimmune encephalomyelitis in mice by modulating inflammatory responses and the gut microbiota

BackgroundAnemoside B4 (AB4) is a representative component of Pulsatilla decoction that is used in traditional Chinese medicine for treating inflammatory conditions. It is not known whether AB4 has beneficial effects on multiple sclerosis (MS). MethodsIn the present study, we examined the preventative and therapeutic effects of AB4, and the possible mechanism by which it protects female mice against experimental autoimmune encephalomyelitis (EAE). ResultsPreventative treatment with AB4 (given orally at 100 and 200 mg/kg for 18 days) reduced the clinical severity of EAE significantly (from 3.6 ± 1.3 to 1.8 ± 1.5 and 1.6 ± 0.6, respectively), and inhibited demyelination and inflammatory infiltration of the spinal cord. In the therapeutic protocol, oral administration of 200 mg/kg AB4 for 21 days after initiation of EAE significantly alleviated disease severity (from 2.6 ± 1.3 to 0.9 ± 0.6) and was as effective as the clinically used drug fingolimod (0.3 ± 0.6). Furthermore, both doses of AB4 significantly inhibited mRNA expression of TNF-α, IL-6, and IL-17, and STAT3 activation, in the spinal cord; and the ex vivo and iv vitro AB4 treatment markedly inhibited secretion of the three cytokines from lymphocytes of EAE mice upon in vitro restimulation. In addition, AB4 reversed the changes in the composition of the intestinal microbiome observed in EAE mice. ConclusionWe reveal for the first time that AB4 protects against EAE by modulating inflammatory responses and the gut microbiota, demonstrating that AB4 may have potential as a therapeutic agent for treating MS in humans.

Detergent exposure induces epithelial barrier dysfunction and eosinophilic inflammation in the esophagus.

Eosinophilic esophagitis (EoE) is a chronic allergic disease associated with type 2 inflammation and epithelial barrier dysfunction. The etiology is unknown, however, genetic heritability studies suggest environmental factors play a key role in pathogenesis. Detergents, such as sodium dodecyl sulfate (SDS), are common ingredients in household products such as dish soap and toothpaste. We hypothesized detergent exposure decreases epithelial barrier function and induces esophageal inflammation. Immortalized esophageal epithelial cells (EPC2) were cultured in air-liquid interface (ALI) and exposed to SDS. Barrier function/activity was assessed by transepithelial electrical resistance (TEER), FITC-dextran flux, and RT-PCR. Additionally, SDS-treated mouse esophageal organoids were evaluated for morphology. To investigate the effects of SDS in vivo, mice were treated with 0.5% SDS in drinking water for 14 days. Esophagi were assessed by gross morphology, histopathology, protein expression, and bulk RNA sequencing. When EPC2 cells were exposed to SDS (5μg/ml) for 96 h, TEER decreased (p=0.03), and FITC-dextran flux increased (p=0.0002). mRNA expression of IL-33 increased 4.5-fold (p=0.02) at 6h and DSG1 decreased (p < 0.0001) by 72 h. Disrupted epithelial integrity was noted in SDS-treated esophageal organoids. When mice were exposed to SDS, they showed increased esophageal width, chemokine, and metalloprotease levels. Mice treated with SDS also showed increased IL-33 protein expression, basal zone hyperplasia, CD4+ cell infiltration, and esophageal eosinophilia. RNA sequencing revealed upregulation of immune response pathway genes. Exposure to SDS decreases esophageal barrier integrity, stimulates IL-33 production, and promotes epithelial hyperplasia and tissue eosinophilia. Detergents may be a key environmental trigger in EoE pathogenesis.