IL-6 Content Research Articles

Effect of Gardeniae Fructus Powder on Growth Performance, Antioxidant Capacity, Intestinal Barrier Function, and Colonic Microbiota of Weaned Piglets

The present study aimed to explore the effect of GF powder on the growth performance, diarrhea rate, antioxidant and immune capacity, and intestinal health of weaned piglets. A total of 144 weaned piglets (8.29 ± 0.11 kg) at 21 d old were randomly assigned to four groups, with each treatment consisting of six replicate pens, with six piglets per pen, and each pen containing three barrows and three gilts. The piglets were fed a basal diet supplement with 0%, 0.4%, 0.6%, and 0.8% GF powder (n = 36). Our results indicated that compared with the basal diet, the F/G and diarrhea rate were remarkably decreased in the 0.8% GF group (p < 0.05). Serum biochemical parameters showed that supplementation with GF significantly increased the content of HDL-C (0.6 and 0.8% levels), IL-6 (0.8% level), IL-10 (0.4, 0.6, and 0.8% levels), Ig G (0.4% level), and Ig A (0.8% level) compared with the basal diet (p < 0.05). The index of antioxidant capacity showed that compared with a basal diet, supplementation with GF significantly decreased serum MDA content (0.4% and 0.8% levels) and jejunal and ileal MDA content (0.4%, 0.6%, and 0.8% levels) (p < 0.05). Additionally, compared with the basal diet, supplementation with GF significantly increased serum and ileal T-AOC content (0.4%, 0.6%, and 0.8% levels), serum T-SOD content (0.4% and 0.8% levels), ileal T-SOD content (0.4%, 0.6%, and 0.8% levels), CAT content (0.4%, 0.6%, and 0.8% levels), and jejunal GSH-Px content (0.8% level) (p < 0.05). The results of gene expression indicate that compared with the basal diet, supplementation with GF significantly increased Nrf 2 (0.4% level), NQO (0.4% level), SOD 1 (0.4% and 0.8% levels), and GCLC (0.4% level) and GCLM (0.8% level) abundance in jejunal mucosa; supplementation with GF significantly increased Nrf 2 (0.4%, 0.6%, and 0.8% levels), HO-1 (0.4% level), NQO (0.8% level), SOD 1 (0.4% and 0.8% levels), and GCLC (0.4% level) and GCLM (0.8% level) abundance in ileal mucosa (p < 0.05). Ulteriorly, the present results indicate that supplementation with GF at the 0.8% level significantly increased the villus height in the jejunum and ileum as well as the villus/crypt ratio in the ileum compared with the basal diet (p < 0.05). Compared with the basal diet, 0.4% GF significantly increased Occludin gene expression in ileal mucosa (p < 0.05), 0.6% GF significantly increased ZO-1, Claudin-1, and Occludin gene expression in jejunal mucosa (p < 0.05), and 0.8% GF significantly increased ZO-1 and Occludin gene expression in jejunal mucosa along with Occludin expression in ileal mucosa (p < 0.05). Furthermore, colonic microbiota composition showed that Shannon, observed species, and Chao 1 indices were significantly increased in the 0.8% GF group compared with the basal diet (p < 0.05). At the phylum level, in comparison with the basal diet, the relative abundance of Firmicutes significantly decreased in the 0.4%, 0.6%, and 0.8% GF groups, and Bacteroidetes increased in the 0.8% GF group (p < 0.05). At the genus level, compared with the basal diet, 0.6% and 0.8% GF significantly increased Prevotella abundance, and 0.6% GF significantly decreased Coprococcus abundance (p < 0.05). At the species level, compared with the basal diet, 0.8% GF significantly increased Prevotella copri abundance, and 0.4%, 0.6%, and 0.8% GF significantly decreased Blautia obeum abundance (p < 0.05). In summary, a dietary supplement with 0.8% Gardeniae Fructus powder significantly decreased the F/G and diarrhea rate and improved antioxidant capacity and intestinal barrier function, which may be associated with the improvement of the relative abundance of Prevotella copri. These findings indicate that Gardeniae Fructus powder may be used as a feed additive in swine weaning.

Effects of Dietary Nano-Composite of Copper and Carbon on Antioxidant Capacity, Immunity, and Cecal Microbiota of Weaned Ira White Rabbits

This experiment investigated the effects of dietary supplementation with nano-composites of copper and carbon (NCCC) on antioxidants, immune functions, and the cecum microbiota of weaned Ira white rabbits. A total of 240 weaned 35-day-old Ira white rabbits were randomly allocated to five dietary treatments (n = 6 per treatment, each replicate consisted of eight rabbits) that included the control group (CON) with a basal diet, the SAL group with 60 mg/kg salinomycin (SAL) in addition to the basal diet, and the NCCC I, II, III groups, which were supplemented with 50, 100, and 200 mg/kg NCCC, respectively, in addition to the basal diet. The test lasted for 28 d. The results showed that dietary NCCC supplementation increased the liver Cu/Zn-SOD content and up-regulated the gene expression of Cu/Zn-SOD (p < 0.05), while also reducing the content of MDA in the liver and enhancing the antioxidant capacity of Ira white rabbits. Moreover, the NCCC diet supplementation reduced the content of IL-6 and down-regulated the relative expression of IL-6 and IL-1β genes in the jejunum of Ira white rabbits (p < 0.05). In addition, the metagenomic analysis of 16 S rRNA showed significant differences in the cecal microbial structure of weaned Ira white rabbits in the NCCC III group compared with the CON, NCCC I, and NCCC II groups (p < 0.05). Firmicutes and Bacteroidetes were the dominant phyla of cecal microorganisms in weaned Ira rabbits in the NCCC diet groups. The dominant genera included unidentified Eubacteriaceae, unclassified Lachnospiraceae, Christensenellaceae, and Ruminococcus. Furthermore, the relative abundance of Ruminococcus in the NCCC I and II groups was lower than that in the CON group in the cecum of Ira white rabbits (p < 0.05). In summary, our results showed that diet supplementation with NCCC could enhance the antioxidant capacity in the liver, alleviate intestinal inflammation, and regulate the structure of intestinal flora, improving the health of Ira white rabbits.

Early Weaning Impairs the Growth Performance of Hu Lambs Through Damaging Intestinal Morphology and Disrupting Serum Metabolite Homeostasis.

This study aimed to evaluate the effect of early weaning (EW) on the growth performance, gastrointestinal development, serum parameters, and metabolomics of Hu sheep lambs. Twenty-four male Hu lambs were initially ewe-reared. A total of 12 lambs were weaned at 30 d of age (D30) as the EW group, and the remaining 12 lambs were weaned at 45 d of age (D45) as the control (CON) group. Serum samples were collected from six lambs per treatment on D30, D33, D36, and D45, and the lambs were slaughtered on D45 to collect the rumen and small intestine. The results showed that, compared with the CON group, the average daily gain (ADG), final body weight (p < 0.001), as well as average daily feed intake (ADFI) of lambs in the EW group significantly decreased in the first (p = 0.004) and second (p = 0.013) 5 days of treatment. Additionally, EW increased the ruminal weight and papillae length but reduced the duodenal villus height on D45 (p < 0.05). As for the serum parameters, the concentrations of glucose on D33, D36, and D45 (p < 0.001), and the IL-6 content on D45 (p = 0.018) were observed to be lower, while the levels of immunoglobulin A (IgA) (p = 0.027), IgG (p = 0.035), and IgM (p = 0.002) on the four ages were all higher in the EW group than those in CON group. Additionally, both treatment and age interactively affected the levels of GLU (p = 0.001), TP (p = 0.041), and IL-6 (p = 0.016). Additionally, the serum metabolomics analysis on D45 showed that the contents of 5-HT and arachidonic acid were increased, while L-phenylalanine, L-tyrosine, and L-glutamic acid were reduced in the EW group (p < 0.05). These differential metabolites were enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including inflammatory mediator regulation, protein digestion and absorption, and phenylalanine and tyrosine biosynthesis. The current results identify that EW at D30 decreased the growth performance (ADG and ADFI) of Hu lambs within two weeks post-weaning, which might be associated with impaired duodenal morphology and glucose metabolism. The serum metabolomics analysis revealed that EW altered the concentrations of 5-HT, phenylalanine, tyrosine, and arachidonic acid, which could serve as potential regulatory targets for modulating the health of EW Hu lambs.

Effect of electroacupuncture on blood-brain barrier and central inflammatory response in obese rats

To observe the effect of electroacupuncture (EA) on central nervous inflammatory response in obese rats, so as to explore its mechanisms underlying improvement of obesity. Male SD rats were randomly allocated to normal control, model, EA and sham-EA groups (n=15/group). Rats of the normal group were fed with normal diet, and the obesity model was established by feeding the rats with high-fat diet for 8 weeks. EA (2 Hz, 1 mA) was applied to "Zusanli"(ST36), "Zhongwan"(CV12), "Guanyuan"(CV4) and "Fenglong"(ST40), and non-acupuncture points (about 5 mm left to CV4 and CV12 and about 5 mm lateral to ST36 and ST40) for 15 min, once every other day, 3 times a week for 8 weeks (no electrical current was delivered to the non-acupoints). The body weight and anal-nose length in each group were recorded before and after the intervention and Lee's index was calculated. The adipocyte morphology of epididymis was detected after H.E. staining, and the contents of serum tumor necrosis factor α (TNF-α), interleukin (IL)-6, IL-10 and free fatty acid (NEFA) were detected by using enzyme-linked immunosorbent assay (ELISA). The expression levels of TNF-α, IL-6, IL-10 and inducible nitric oxide synthase (iNOS) proteins in the hypothalamus tissue were detected by Western blot, and the immunoactivity of ionized calcium-binding adapter molecule 1 (Iba-1) in hypothalamic microglia was detected by immunohistochemistry. The content of Evans blue in the brain tissue samples was detected by Evans blue assay to evaluate the integrality of blood-brain barrier. Compared with the normal group, the body mass (BM), Lee's index, contents of serum TNF-α, IL-6 and NEFA, expression levels of hypothalamic TNF-α, IL-6 and iNOS proteins, the immunoactivity level of Iba-1 and the content of Evans blue were significantly increased (P<0.01, P<0.05), while the content of serum IL-10 and the expression of hypothalamic IL-10 protein considerably decreased (P<0.01) in the model group. After the intervention, modeling induced increase of BM, Lee's index, contents of serum TNF-α, IL-6 and NEFA, expression levels of hypothalamic TNF-α, IL-6 and iNOS proteins, the immunoactivity level of Iba-1 and the content of Evans blue, and decrease of serum content and expression level of IL-10 in the hypothalamus were reversed by EA stimulation (P<0.01, P<0.05) but not by non-acupoint stimulation. H.E. staining showed that the adipocytes of epididymis became larger in size and decreased in the number in the model group relevant to the normal group, and became smaller after EA (not non-acupoint) relevant to the model group. EA can treat obesity, improve body inflammation caused by obesity, and relieve central inflammation by repairing the blood-brain barrier.

Electroacupuncture improves pulmonary function by reducing inflammatory reaction via inhibiting miR-19b-3p to regulate SOCS3/JAK1/STAT3 signaling pathway in mice with chronic obstructive pulmonary disease

To explore whether electroacupuncture(EA) can increase the expression of suppressor of cytokine signaling (SOCS)3 by affecting the expression of miR-19b-3p, inhibiting the continuous activation of janus kinase (JAK)1 /signal transducer and activator of transcription (STAT)3 signaling pathway, and improve pulmonary inflammation in chronic obstructive pulmonary disease (COPD) mice. Forty mice were randomly divided into normal, COPD model, COPD+EA, and COPD+miR-19b-3p agomir (agomir)+EA groups. The COPD model was simulated by cigarette smoke exposure for 1 h, twice a day for 3 months. After modeling, mice in the COPD+EA group and the COPD+agomir+EA group received EA stimulation of "Feishu" (BL13) and "Zusanli"(ST36) for 30 min, once every other day, for 14 days. The mice of the COPD+agomir+EA group received intranasal drops of miR-19b-3p agomir solution (50 μL) 24 h before every EA intervention. The pulmonary ventilation functions including forced vital capacity (FVC), forced expiratory volume (FEV) at the 0.05 s and 0.1 s (FEV0.05 and FEV0.1), FEV0.05/FVC and FEV0.1/FVC were detected using a pulmonary function analysis system. The pathological morphology of lung tissue was observed after H.E. staining. The contents of interleukin(IL)-6, tumor necrosis factor(TNF)-α and IL-1β in the bronchoalveolar lavage fluid were assayed using ELISA. And the expressions of SOCS3, JAK1, STAT3, phosphorylated(p)-JAK1 and p-STAT3 proteins in the lung tissue were detected using Western blot. The expressions of miR-19b-3p, JAK1, STAT3 and SOCS3 mRNA were detected using real-time fluorescence quantitative PCR. Compared with the normal group, the COPD model group had a significant decrease in the levels of FVC, FEV0.05, FEV0.1, FEV0.05/FVC and FEV0.1/FVC, and the expression levels of SOCS3 mRNA and protein (P<0.001) and a significant increase in the contents of IL-6, TNF-α and IL-1β, and the expression levels of miR-19b-3p, JAK1 and STAT1 mRNAs, and p-JAK/JAK1 and p-STAT3/STAT3 protein expression ratio (P<0.001). After EA intervention, the decreased and increased levels of all the indexes mentioned above were reversed in the COPD+EA group (P<0.001, P<0.01, P<0.05). The effect of the EA+agomir were significantly less than EA in up-regulating the levels of FVC, FEV0.05, FEV0.1, FEV0.05/FVC and FEV0.1/FVC, and the expression levels of SOCS3 protein and in down-regulating the contents of IL-6, IL-1β, TNF-α, and expressions of JAK1 and STAT3 mRNA, and p-STAT3/STAT3 (P<0.001, P<0.01, P<0.05), suggesting that the effects of EA were weakened after intranasal drops of miR-19b-3p agomir. H.E. staining showed thickened alveolar wall, obvious inflammatory cell infiltration, with some ruptured alveoli fused into large vesicles in the model group, slightly dilated alveoli and small amount of inflammatory cell infiltration in the COPD+EA group, and slight alveolar fusion, slight thickening of alveolar wall, and light inflammatory cell infiltration in the COPD+agomir+EA group. EA can inhibit the expression of miR-19b-3p, thereby up-regulating SOCS3 expression and inhibiting the overactivation of JAK1/STAT3 signaling, thus reducing lung inflammatory reaction to improve pulmonary function in mice with COPD.

Indoleamine 2, 3-dioxygenase Regulates the Differentiation of T Lymphocytes to Promote the Growth of Gastric Cancer Cells through the PI3K/Akt/mTOR Pathway.

To investigate the regulatory mechanism of indoleamine 2, 3-dioxygenase (IDO) in T lymphocyte differentiation and its role in promoting the growth of gastric cancer (GC) cells through the PI3K/Akt/mTOR pathway. GC cell lines (MFC and NCI-N87) and PBMC cells were co-cultured and IDO inhibitor 1-methyl-tryptophan (1-MT) was added. The proliferation was detected by CCK-8, the apoptosis was detected by flow cytometry, and the contents of TNF-α, IL-1β, IL-6, IL-8, and INF-γ were detected by ELISA. The expression levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, and p-mTOR were tested using Western blot, and the proportion of CD4+/CD8+, CD4+CD25+Foxp3+Treg cells was detected by flow cytometry. C57BL/6 mice were used to establish the MFC GC mouse model and treated with 1-MT. The changes in body weight and tumor diameter were measured. Ki-67, CD4+, CD8+, and CD25+ expressions were detected by immunohistochemistry. IDO promoted the proliferation of MFC and NCI-N87 cells, inhibited apoptosis, and decreased the levels of TNF-α, IL-1β, IL-6, IL-8, and INF-γ in the supernatant after co-culture with BPMC. The expressions of p-AKT, p-mTOR, and p-PI3K increased after 1-MT treatment. The proportion of CD4+/CD8+ cells was increased and the proportion of Treg cells was decreased in PBMC cells after the addition of 1-MT. Overexpression of IDO suppressed T cells differentiation by inhibiting the PI3K/Akt/mTOR pathway. In vivo, 1-MT treatment reduced the tumor size and weight, increased CD4+ and CD8+ positive area proportion, and decreased Ki-67 and CD25+ positive area proportion. Co-culture of GC cells and immune cells promotes the proliferation of GC cells and inhibits apoptosis, which can be reversed by 1-MT. IDO may suppress the proliferation of T lymphocyte through inhibiting the PI3K/Akt/mTOR signaling pathway. This provides new evidence for the potential of exploiting IDO inhibitors for GC treatment.

Serum adipocytokines and inflammatory cytokines in pregnant women with gestational diabetes mellitus: clinical utility and development of a risk prediction model

ABSTRACT Background This study analyzed the clinical utility of serum adipocytokines and inflammatory cytokines in gestational diabetes mellitus (GDM) and developed a quantitative nomogram prediction model. Research Design & Methods General data were collected. Fasting venous blood was taken and levels of fasting plasma glucose (FPG), serum adipocytokines, and inflammatory cytokines were assessed. The main risk factors for GDM were analyzed by implementing univariate and multivariate logistic regression analysis. The weights of the main risk factors were assigned, and the nomogram prediction model for GDM was developed by R software. The efficacy of the nomogram model for GDM prediction was measured and analyzed by the receiver operating characteristic (ROC) curve and calibration curve. Results The observation group possessed a higher proportion of family history of diabetes, raised FPG, LEP, Visfatin, hs-CRP, IL-6, and TNF-α contents, and lower ADP contents (all p < 0.05). Multivariate logistic regression analysis displayed that LEP, ADP, and IL-6 were the main risk factors for GDM (p < 0.05). Calibration curve was basically consistent with the original curve, suggesting good accuracy. Conclusion Serum adipocytokines and inflammatory cytokines were the main risk factors for GDM. Developing a nomogram model can facilitate early diagnosis of GDM by physicians, allowing for timely interventions.

Study on the mechanism of moxibustion regulating ferroptosis-lipid metabolism pathway to improve synovitis inflammatory injury in rheumatoid arthritis rats

To explore the mechanism of moxibustion in improving synovitis inflammatory injury in rheumatoid arthritis (RA) model rats based on the ferroptosis-lipid metabolism pathway. Forty-eight SD rats were randomly divided into normal, model, moxibustion and moxibustion + long-chain acyl-CoA synthetase 4 (ACSL4) inhibitor groups, with 12 rats in each group. The RA model was replicated using environmental factors of wind, cold, and dampness combined with Freund's complete adjuvant injection. The moxibustion group received moxibustion at "Shenshu" (BL23) and "Zusanli" (ST36) for 20 minutes per acupoint each time, once daily at a single acupoint (both sides) with alternating acupoints over 15 consecutive days. The moxibustion + ACSL4 inhibitor group received intraperitoneal injection of the ACSL4 inhibitor rosiglitazone (0.4 mg/kg) after moxibustion, once daily for 15 consecutive days. Histopathological changes in synovial tissue were observed using HE staining；serum contents of glutathione (GSH) and malondialdehyde (MDA) were detected using biochemical methods；reactive oxygen species (ROS) levels in synovial tissues were detected using flow cytometry；the expressions of glutathione peroxidase 4 (GPX4), ACSL4, and lysophosphatidylcholine acyltransferase 3 (LPCAT3) proteins in synovial tissues were detected using Western blot；and serum contents of interleukin (IL)-6 and tumor necrosis factor-alpha (TNF-α) were detected using ELISA. Compared with the normal group, the serum contents of GSH decreased (P<0.01) while MDA, IL-6, and TNF-α contents increased (P<0.01)；the ROS levels, ACSL4 and LPCAT3 protein expressions increased (P<0.01) while GPX4 protein expression decreased (P<0.01) in synovial tissue in the model group. Compared with the model group, the serum contents of GSH increased (P<0.01) while MDA, IL-6, and TNF-α contents decreased (P<0.01)；the ROS levels, ACSL4 and LPCAT3 protein expressions decreased (P<0.01) while GPX4 protein expression increased (P<0.01) in synovial tissue in the moxibustion group and moxibustion + ACSL4 inhibitor group. Compared with the moxibustion group, the serum contents of GSH increased (P<0.01) while MDA, IL-6, and TNF-α contents decreased (P<0.01)；the ROS levels, ACSL4 and LPCAT3 protein expressions decreased (P<0.01, P<0.05) while GPX4 protein expression increased (P<0.05) in synovial tissue in the moxibustion + ACSL4 inhibitor group. HE staining showed that the model group had significantly increased synovial hyperplasia and inflammatory cell infiltration；the moxibustion group and moxibustion + ACSL4 inhibitor group showed varying degrees of alleviation in inflammatory cell infiltration and hyperplasia in synovial tissue；compared with the moxibustion group, the moxibustion + ACSL4 inhibitor group showed more significant improvements in inflammatory infiltration and hyperplasia of synovial cells, reduced layers of synovium. Moxibustion at BL23 and ST36 can alleviate synovial inflammatory injury, and its mechanism may be related to reducing lipid peroxidation and ROS levels, and inhibiting the occurrence of ferroptosis.

Ameliorative effect of phenolic compound-pterostilbene on corticosterone-induced hepatic lipid metabolic disorder in broilers

The aim of this study was to investigate the ameliorative effects of pterostilbene (PTE), a polyphenolic compound, on stress-induced lipid metabolic disorders in the liver of broiler chickens. Six hundred healthy, one-day-old Arbor Acres (AA) with similar weight were randomly assigned to five groups, each consisting of 8 replicates with 15 broilers per replicate. The groups included: a control group (fed a basal diet), and four groups treated with corticosterone (CORT) at varying dietary levels of PTE supplementation: CORT (0 mg/kg PTE), CORT-PT200 (200 mg/kg PTE), CORT-PT400 (400 mg/kg PTE), and CORT-PT600 (600 mg/kg PTE). The results indicated that PTE administration to corticosterone (CORT)-injected broilers significantly improved weight gain, reduced liver index, and lowered the elevation of serum AST, GGT, Glu, TC, TG, and LDL-c concentrations induced by CORT injection (P < 0.05), but had no significant effect on serum CORT concentration (P > 0.05). PTE also significantly reduced the increased rate of abdominal fat deposition induced by CORT, decreased the average size of adipocytes, and downregulated the expression of the FAS gene (P < 0.05). It reversed the increase in liver TC, TG, LDL-c, and NEFA content induced by CORT (P < 0.05). PTE had no significant effect on the expression of the glucocorticoid receptor GR (P > 0.05), but significantly upregulated the protein expression of Sirt1 and p-AMPK (P < 0.05), promoted the expression of lipid autophagy genes MAP1LC3B and lipolytic genes LPL, but inhibited the expression of fatty acid synthesis genes SREBP-1c, ACC, and SCD (P < 0.05). The addition of PTE to the diet alleviated CORT-induced oxidative stress and inflammation by enhancing T-SOD and GSH-Px activities, reducing MDA content, inhibiting p-NF-κB p65 and NLRP3 expression and the release of TNF-α and IL-1β in the serum, and increasing IL-4 content (P < 0.05). Overall, dietary PTE effectively regulates lipid metabolism and antioxidant status, offering a potential strategy to mitigate stress-induced metabolic disruptions in broilers.

CHANGES IN TRYPTOPHAN METABOLISM AND CYTOKINE CONCENTRATION IN SYNOVIAL JOINT TISSUE IN RHEUMATOID ARTHRITIS

The aim of the research. To identify changes in tryptophan and cytokine metabolism in synovial joint tissue in rheumatoid arthritis. Research materials and methods. The experiments were performed on 40 WISTAR rats. At the beginning of the study, each animal of the experimental groups received an intraperitoneal injection of collagen type 2 solution (Chondrex, Inc., USA) in Freund's incomplete adjuvant. The content of tryptophan, kynurenine, 3-hydroxykinurinine, L-5-hydrotryptophan by HPLC with fluorimetric and spectrophotometric detection, and the concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa were determined in the tissues of the synovial membrane of the knee joint. They were determined using Rat Inflammation Panel V02 multiplex assay kits (Biolegend, USA) on a Cytomics FC500 flow cytofluorimeter (Beckman Coulter, USA). Statistical analysis was performed using the Jamovi program version 2.3.Results. The concentration of cytokines: IL1b, IL6, IL10, IL17, IL18, TNFa was found to be high during the acute development of rheumatoid arthritis. With experimental rheumatoid arthritis, the content of tryptophan metabolites along the kynurenine pathway increases and the concentration of metabolites along the serotonin pathway decreases. Direct positive correlations of IL1b, IL6, IL10, IL17, IL18, TNFa with the content of tryptophan metabolites along the kynurenine pathway have been established.Conclusions. Tryptophan metabolism and IL1b, IL6, IL10, IL17, IL18, and TNFa content are altered in rheumotoid arthritis.

Exploring the Effect and Mechanism of DaYuan Yin Against Acute Lung Injury by Network Pharmacology, Molecular Docking, and Experimental Validation.

DaYuan Yin (DYY), a traditional Chinese medicine for lung diseases, requires further study to understand how it improves acute lung injury (ALI). This study seeks to elucidate the material basis and molecular mechanisms underlying the treatment of ALI with DYY through network pharmacology, molecular docking, and experimental validation. DYY's active components and targets were identified using TCMSP and UHPLC-MS/MS, and a herb-component-target network was created with Cytoscape 3.7.2. ALI target genes were sourced from GeneCards, DisGeNET, and DrugBank. A PPI network was built, with core targets analyzed through GO and KEGG enrichment via Metscape. The therapeutic effects and mechanisms of DYY on LPS-induced ALI in rats were explored, and molecular docking evaluated the interactions between Nrf2, HO-1, TLR4, and the components. The study identified 95 active compounds, 234 therapeutic targets, and 2529 ALI-related genes, with 111 shared targets between DYY and ALI. KEGG analysis indicates that the PI3K-AKT, MAPK, and oxidative stress pathways are associated with DYY's anti-ALI effects. Network pharmacology and UHPLC-MS/MS analysis revealed active ingredients like quercetin, Magnolol, and Wogonin. Compared with the model group, DYY reduced the lung dry-wet ratio (W/D) of ALI rats from (5.31 ± 0.51) to (4.47 ± 0.73)(P < 0.05). Meanwhile, the contents of IL-6 and TNF-α in bronchoalveolar lavage fluid (BALF) and MDA, NO and ROS in lung tissue were also significantly decreased. Notably, DYY enhances UCP2 mRNA expression, boosts Nrf2 and HO-1 expression, and inhibits TLR4-mediated pro-inflammatory mediators. Molecular docking analysis showed that the main components of DYY had strong binding ability with HO-1. DYY can alleviate inflammation, oxidative stress, and ALI-related changes by targeting the Nrf2/HO-1 mediated TLR4 pathway, providing insights for developing effective ALI treatments.

Arsenic exposure accelerates type 1 diabetes mellitus progression via pyroptosis pathway in mice.

The relationship between arsenic exposure and the development of diabetes mellitus has garnered significant interest in recent years. However, current experimental studies have not definitively established the role of arsenic in the onset of diabetes mellitus. To investigate this relationship specifically concerning type 1 diabetes mellitus, Streptozocin (STZ) was utilized as an inducer to initiate the fundamental pathological changes associated with the disease. A high dose of STZ (50mg/kg) served as the positive control, while a low dose of STZ (20mg/kg) was administered in combination with arsenic at varying doses. The objective was to determine whether arsenic enhances the effects of STZ, thereby leading to an expedited onset and progression of type 1 diabetes mellitus. The preliminary investigation into the impact of arsenic exposure on experimental type 1 diabetic mice focused on the NLRP3/Caspase-1/GSDMD mediated pyroptosis pathway. The results showed that fasting blood glucose (FBG) was increased, glucose tolerance was impaired, insulin sensitivity was decreased, fasting serum insulin and the homeostatic model assessment-β (HOMA-β) were significantly reduced, hair arsenic content was increased, reactive oxygen species(ROS), interleukin (IL)-1β and IL-18 contents were increased, and the pathological morphology of pancreas was more serious in the combined group. Moreover, the expression levels of proteins associated with the NLRP3/Caspase-1/GSDMD-mediated pyroptosis pathway were elevated in the combined group. This study illustrates that exposure to arsenic, combined with low-dose STZ, not only leads to pancreatic injury in mice, impacting insulin secretion and causing elevated blood glucose levels, thereby hastening the progression of type 1 diabetes, but also induces pyroptosis in pancreatic tissues by influencing the NLRP3/Caspase-1/GSDMD signaling pathway, further facilitating the development of type 1 diabetes.

Application of Lactobacillus plantarum and Pediococcus lactis on Lipid Metabolism, Anti-Inflammatory, and Fecal Microbiota in Cats.

Probiotics have been used in functional foods and dietary supplements, and in recent years, they have become more widely used in pets. In our previous experiment, Lactobacillus plantarum L-27-2 and Pediococcus lactis L-14-1 were isolated from cat feces and proved to have positive effects on lipid metabolism in mice. To further discuss their possible effects in cats, a total of 12 healthy cats (British Shorthair) were randomly divided into two groups. One group was fed Pediococcus lactis L-14-1 (1 × 109 CFU/kg/d, n = 6), and the other group was fed Lactobacillus plantarum L-27-2 (1 × 109 CFU/kg/d, n = 6), and the experiment was conducted for 28 days. Blood and feces were collected on days 0 and 28 separately. ELISA was used to detect blood biochemical indexes in cats. The results showed that L-27-2 and L-14-1 could reduce the content of TG (triglyceride, p < 0.05) and LDL-C (low-density lipoprotein cholesterol, p < 0.01) in the blood, increase the content of HDL-C (high-density lipoprotein, p < 0.01), and L-27-2 could significantly reduce the content of IL-6 (p < 0.01). The diversity of feces microbiota was also tested. On the phylum level, there was no significance in the phylum level of Firmicutes and Bacteroidetes (p > 0.05), but on the genus level, in the L-14-1 group, the abundance of Lantiplantibacillus and Cetobacterium was increased (p < 0.05), and the abundance of Ruminococcus, Olsenella, and Labanicoccus was decreased (p < 0.05), while in the L-27-2 group, the abundance of Libanicoccus was also decreased in L-14-1 (p < 0.05). Above all, L-27-2 and L-14-1 can be considered potential probiotics to improve cat gut health and lipid metabolism.

Acupoint injection inhibiting abnormal secretion of mucin and inflammatory reaction of nasal mucosa in rats with allergic rhinitis through the p38 mitogen-activated protein kinase pathway

To observe the effects of acupoint injection on the expression of p38 mitogen-activated protein kinase (MAPK), phosphorylated (p)-p38 MAPK, mucin 5 subtype AC (MUC5AC) in the nasal mucosa, and serum inflammatory factors of rats with allergic rhinitis, so as to explore the mechanism of acupoint injection in improving inflammatory reactions in the nasal mucosa of rats with allergic rhinitis. SD rats were randomly divided into the normal group, model group, acupoint injection group, and non-acupoint group, with 7 rats in each group. The allergic rhinitis rat model was established using ovalbumin sensitization. Intervention was performed by injecting dexamethasone and 1% lidocaine mixture (0.05 mL) into the bilateral "Yingxiang"(LI20) and "Yintang"(GV24+) acupoints, with injection at non-acupoints for non-acupoint group as the control, once every 3 days for a total of 4 times. Allergic symptoms in the rat nose were evaluated using a cumulative scoring method；histopathological changes in the nasal mucosa were observed after HE staining；serum histamine (HA), interleukin (IL)-6, IL-8, and tumor necrosis factor-alpha (TNF-α) contents were detected using ELISA；real-time fluorescent quantitative PCR was used to detect the expression of MUC5AC mRNA in the nasal mucosa；Western blot was used to detect the expression of p38 MAPK, p-p38 MAPK, and MUC5AC protein in the nasal mucosa. Compared with the normal group, nasal allergic symptom score, serum HA, IL-6, IL-8, and TNF-α contents, nasal mucosal p38 MAPK, p-p38 MAPK proteins expression, MUC5AC mRNA and protein expression were significantly increased(P<0.01) in the model group. Compared with the model group and non-acupoint group, nasal allergic symptom score, serum HA, IL-6, IL-8, and TNF-α contents, nasal mucosal p38 MAPK and p-p38 MAPK proteins expression, MUC5AC mRNA and protein expression were significantly reduced (P<0.01, P<0.05) in the acupoint injection group. Acupoint injection can reduce nasal allergic symptom, and MUC5AC secretion in rats with allergic rhinitis, alleviate nasal mucosal inflammatory reactions, and its mechanism of action may be related to the inhibition of p38 MAPK phosphorylation, thereby reducing the release of inflammatory factors.

Effects of electroacupuncture on expression of miR-218 and HMGB1/TLR4 signaling pathway in rats with focal cerebral ischemia

To observe the effect of electroacupuncture (EA) on high mobility group box 1 (HMGB1) / toll-like receptor 4 (TLR4) signaling pathway and inflammatory injury in rats with focal cerebral ischemia (FCI), so as to explore its mechanisms underlying amelioration of ischemic stroke. Male SD rats were randomly divided into sham operation (n=18), model (n=18) and EA (n=18) groups. The FCI model was established according to Longa's method. In the EA group, 24 h after modeling, EA stimulation was applied to "Baihui"(GV20), "Fengfu"(GV16), and "Neiguan" (PC6) and "Xinshu" (BL15) on the left side for 20 min, once a day for 1 week. The neurological deficit severity was evaluated by the modified neurological severity score (mNSS), and the morphological changes of neurons were observed after H.E. staining. The expression of HMGB1 and TLR4 proteins were detected by immunofluorescence staining. The contents of tumor necrosis factor(TNF)-α and interleukin(IL)-6 in the ischemic area tissue of the brain were detected using ELISA. The mRNA expressions of HMGB1 and TLR4 as well as the expression of miR-218 in the ischemic cortex were detected by PCR. Compared with the sham operation group, the mNSS at 2 h, 48 h and 7 day (d), IL-6 and TNF-α contents, HMGB1 and TLR4 immunoactivity and mRNA expression at 48 h and 7 d were significantly increased (P<0.01), while miR-218 expression at 48 h and 7 d was obviously down-regulated (P<0.01) in the model group. In comparison with the model group, the mNSS at 7 d, IL-6 and TNF-α contents, HMGB1 and TLR4 immunoactivity and mRNA expression at 7 d not at 48 h were significantly decreased (P<0.01, P<0.05), while the expression of miR-218 at 7 d was apparently up-regulated (P<0.05) in the EA group. H.E. staining showed that there were signs of neuronal necrosis including blurred outline of the cells, disordered internal structure, dense cytoplasm, shrunk nuclei, etc. in the ischemic area of the brain at 48 h and 7 d in the model group, which was evidently milder on day 7 in the EA group. EA can reduce the inflammatory injury of brain tissue and improve the neurological impairment in rats with FCI, which may be associated with its effects in inhibiting HMGB1/TLR4 signaling and down-regulating the level of pro-inflammatory cytokines.

Muscle Tissue Response to Vipera berus berus and Vipera berus nikolskii Venoms: A Study on Matrix Metalloproteinases and Cytokine Modulation

Vipera berus, also known as the common adder, is a snake species widely distributed in Europe and parts of Asia. Its venom is a mix of enzymatic and non-enzymatic components causing both local and systemic effects including edema, hemorrhage, myonecrosis, gastrointestinal disorders, and rarely – rhabdomyolysis, coagulopathy and hypovolemic shock. Matrix metalloproteinases (MMPs) and cytokines, being inflammatory mediators, participate both in the development of these reactions and in tissue regeneration after acute damage. The current study was aimed at analyzing the concentrations of important mediators of tissue damage, inflammation and regeneration, namely, MMP-1, -2, -3, -8, -10, TIMP-1 (a tissue inhibitor of MMPs), and pro- and anti-inflammatory cytokines, in rat muscle after intraperitoneal injection of venoms from two subspecies of adders – V. berus berus and V. berus nikolskii — to clarify their role in the local response of skeletal muscle to the studied venoms. Changes in muscle tissue were revealed, namely, significant increase in almost all MMPs including MMP-2, as well as TIMP-1, IFN-γ and anti-inflammatory IL-4 and IL-10 content. Moreover, a tendency for some pro-inflammatory cytokines levels to decline 24 h after venom administration was observed, which may be a sign of inflammation suppression and the beginning of reparative processes associated with extracellular matrix remodeling and formation of an optimal environment for muscle regeneration. However, to establish the exact mechanisms of the changes we found, further studies are needed. The ability of V. berus nikolskii venom to induce more pronounced changes was also shown. Our results may be useful for the development of differentiated and more effective treatment strategies for the consequences of bites by these snakes, which include myonecrosis

Efficacy and safety of Shengjiang Xiexin decoction on irinotecan-induced diarrhea in small cell lung cancer patients: a multicenter, randomized, double-blind, placebo-controlled trial

BackgroundIrinotecan is a standard chemotherapeutic agent in small cell lung cancer (SCLC), however, as a common adverse reaction, diarrhea limits the use of irinotecan. Shengjiang Xiexin decoction (SXD) has been used in various gastrointestinal diseases in China two thousand years ago. We designed this clinical trial to supply more evidences on the use of SXD as prophylaxis for irinotecan-induced diarrhea, especially for high-risk population predicted by gene testing of uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1).MethodsIn this clinical trial, 120 patients with SCLC were recruited from six hospitals in China. They received two cycles of chemotherapy, meanwhile they were randomized to receive SXD or placebo for 14 days of oral administration in each cycle of chemotherapy. The primary outcome is the incidence of diarrhea. And secondary outcomes include the the degree of diarrhea and neutropenia, the number of chemotherapy cycles with diarrhea, first occurrence time and duration of diarrhea. To evaluate the effect of SXD on the intestine, a rat model with delayed-onset diarrhea induced by irinotecan was established, and the expression of inflammatory factors including IL-1β, IL-6 and TNF-α, anti-inflammatory factors including IL-10, TGF- β in jejunal tissue was detected by ELISA.Results101 patients (53 in SXD group, 48 in placebo group) completed the trial. The incidence of diarrhea in SXD group and placebo group were 26.42% (14/53) and 52.08% (25/48), respectively (P < 0.05), and the degree of diarrhea also had significant differences (P < 0.05). In UGT1A1 high-risk population, the incidence of diarrhea in two groups were 9.09% and 66.67% (P < 0.05), but there was no significant differences in UGT1A1 low-risk population. The incidence of neutropenia with degree 1–3 between two groups was 20.75% vs 20.83%, 13.21% vs 18.57%, 9.43% vs 20.83% (P < 0.05). No severe adverse events were reported in any group. And animal studies had shown SXD reduced content of IL-1β, IL-6, TNF-α, increased content of IL-10, TGF-β in jejunum tissue.ConclusionsSXD had a prophylactic effect in the diarrhea induced by irinotecan, especially for UGT1A1 high-risk population, and this effect from SXD appeared to be maintained the completion of chemotherapy schedule. The mechanism of action of SXD was related to the regulation of inflammatory factors.Trial registration Chinese Clinical Trial Register: ChiCTR1800018490. Registered on 20 September 2018. https://www.chictr.org.cn/showproj.html?proj=25250. The preliminary protocol of this clinical study has been published in the journal “Trials” in the form of protocol before this paper (Deng et al. in Trials 21:370, 2020).

Serum Concentrations of Pro- and Anti-Inflammatory Cytokines in Wistar Rats Subjected to Immobilization Stress and Treated with Tuftsin-Pro-Gly-Pro Peptide.

It was found that 28-day immobilization stress in male Wistar rats leads to significant increase in the serum concentrations of proinflammatory cytokines IL-1β, IL-6, IFNγ, and monocyte chemoattractant protein-1 (MCP-1) and a decrease in the content of anti-inflammatory IL-10, as well as the ratio IL-1β/IL-10, IL-6/IL-10, and IFNγ/IL-10 compared to non-stressed animals. Administration of the heptapeptide tuftsin-Pro-Gly-Pro produced a corrective effect on cytokine concentrations under stress conditions: at a dose of 750 μg/kg, the peptide decreased the concentrations of IL-1β, IFNγ, and MCP-1 and cytokine ratios (IL-1β/IL-10 and IFNγ/IL-10), while at a dose of 250 μg/kg, it increased IL-10 levels and decreased IL-6/IL-10 and IFNγ/IL-10 ratios.

Efficacy and Mechanisms of Jiyin Fang (Granule) Against Postmenopausal Coronary Heart Disease Complicated with Osteoporosis

Purpose To explore the efficacy and mechanisms of Jiyin Fang (JYF) against postmenopausal coronary heart disease (CHD) complicated with osteoporosis (OP). Methods Firstly, collected ingredients and relative targets of JYF from TCMSP and BATMAN-TCM database, disease targets of CHD and OP from GeneCards, TTD and OMIM database. By use of protein to protein (PPI) network, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, obtained core targets and mechanism pathways. Finally, preliminarily explore the pharmacological effect and major targets partly to explain the mechanism, through experiment of ovariectomized CHD with OP model rats treated with JYF. Results Total of 112 ingredients and 253 related targets of JYF, 173 disease targets common to CHD and OP were screened out. PPI network indicated 9 core targets. GO and KEGG enrichment analysis showed 20 major signal pathway. Rats experiment displayed that JYF could ameliorate the structural injury of aortic arch and femur; reduce the abnormal changes of ST segment and T wave of ECG; decrease the content of BGP, OPG, ALP, CTX, IL-6 and TNFα, increase the levels of PINP in serum; inhibit P53, MAPK, NF-κB and IKK protein expression, promote PPARG protein expression of aortic arch; inhibit NRF2, AKT protein expression and up-regulate p-STAT1, MMP9 protein expression in femur. Conclusion JYF can achieve the purpose of anti postmenopausal CHD complicated with OP. The potential mechanism may relate to P53/MAPK/NF-κB in aortic arch, and AKT/NRF2/p-STAT1 in bone, due to IL-6 and TNFα.

Diallyl trisulfide alleviates dextran sulphate sodium-induced colitis in mice by inhibiting NLRP3 inflammasome activation via ROS/Trx-1 pathway.

Diallyl trisulfide (DATS), a sulphur-containing compound isolated from the medicinal food plant garlic, has been previously reported to attenuate experimental colitis induced by either dextran sodium sulphate (DSS) or 2,4,6-trinitrobenzenesulfonic acid (TNBS) in mice; however, the underlying mechanism remains to be identified. In this study, we deciphered the key mechanism by which DATS alleviates ulcerative colitis (UC). We showed that oral administration of DATS for 10 consecutive days greatly restrained the infiltration of macrophages and the pathological changes in colonic tissues of mice with DSS-induced colitis. DATS treatment notably dampened the content of IL-1β and IL-18 and suppressed NLRP3 inflammasome activation in colon. Mechanistically, DATS effectively diminished the generation of ROS in macrophages. The suppressive effect of DATS on the activation of NLRP3 inflammasome and downregulation of IL-18 and IL-1β levels was blunted by xanthine oxidase. Further studies revealed that DATS inhibited NF-κB pathway activation by suppressing the expression of Trx-1, thereby inhibiting NLRP3 inflammasome activation. Trx-1 overexpression and interference in macrophages promoted and diminished NLRP3 inflammasome activation, respectively. In summary, garlic and its main active ingredient DATS have potentials to prevent and treat UC, and DATS functions by inhibiting NLRP3 inflammasome activation via Trx-1/ROS pathway.