This study aimed to investigate the effects of sodium butyrate (NaB) on plaque formation and inflammatory responses in atherosclerotic (AS) rats through the nucleotide-binding oligodomain-like receptor 1 (NOD1)/receptor-interacting protein 2 (RIP2)/NF-κB signaling pathway. AS model on 41 rats was given by combination of balloon injury and high-fat diet (HFD), and a total of thirty-six rats were successfully modeled. Thirty-six AS rats were divided into model group, NaB group, and NaB+NOD1 agonist γ-D-glu-mesodiaminopimelic acid (iE-DAP) group, with 12 rats in each group. The sham operation group rats were only given exposure of left common carotid artery and successive suture. The model group, NaB group and NaB+iE-DAP group were cultured with HFD while the sham operation group was cultured routinely. NaB group was gavaged with 10 g/(kg·d) NaB, NaB+iE-DAP group was gavaged with 10 g/(kg·d) NaB and peritoneally injected with 1 mL/(each·week) iE-DAP; Model group and sham operation group were gavaged and peritoneally injected with the same volume saline; All the above treatments lasted for twelve weeks. Aorta histomorphology was evaluated by H-E staining. The levels of serum TG, TC, LDL-C, HDL-C, ox-LDL, MCP-1, hs-CRP, TNF-α, and IL-1 were measured by ELISA. Immunohistochemistry was performed to measure ICAM-1 protein expression in the aorta and western blotting was used to measure the protein levels of NOD1, RIP2 and NF-κB in the aorta. The results showed that the aorta lesion was severe in the model group, alleviated in the NaB group and aggravated in the NaB+iE-DAP group. Compared to those of the sham operation group, the serum levels of TG, TC, LDL-C, ox-LDL, MCP-1, hs-CRP, TNF-α,and IL-1, the positive rate of aortic ICAM-1, the expression levels of NOD1, RIP2 and nuclear NF-κB in the model group were significantly increased (all with P<0.05), while the levels of HDL-C and cytoplasmic NF-κB protein were significantly decreased (both P<0.05). In the NaB group, all these indexes showed changes in the reverse direction. Compared to those of the NaB group, the above indicators in the NaB+iE-DAP group all increased significantly while HDL-C and cytoplasmic expression of NF-κB decreased (all with P<0.05). This study suggests that NaB inhibit NOD1/RIP2/NF-κB signaling pathway to relieve plaque formation and inflammatory responses in AS rats.
To explore the effect of matrix metalloproteinase 7 (MMP-7) over-expression on macrophage polarization, galectin 3 (Gal-3) and inflammatory factors secretions, THP-1 cells were induced to macrophages and transfected with MMP-7 over-expressing plasmid or empty vector. The cells were divided into control group, MMP-7 over-expressing group, MMP-7 NC group, M1-polarized group, and M2-polarized group. mRNA expression levels of Gal-3 and MMP-7 were quantified by qPCR. The percentages of F4/80+, CD86+, and CD206+ macrophages were detected by flow cytometry. The expressions of inflammatory cytokines IL-1α, IL-6, and TGF-β were detected by ELISA. After Gal-3 inhibitor treatment on M1-induced and M2-induced macrophages, the mRNA expression of Gal-3 in above macrophages was detected by qPCR, and the levels of IL-1α, IL-6, and TGF-β in the supernatant of cell cultures were detected by ELISA. The results showed that compared to the control group, the MMP-7 over-expressing group and the M2-polarized group showed significant increase in the expressions of Gal-3 and MMP-7 mRNA, in the percentage of (F4/80+)/CD206+ macrophages (all with P<0.01), and in the concentrations of TGF-β(both P<0.01), while IL-1α and IL-6 levels showed significant decrease (both P<0.01). In the M1-polarized group, the percentage of (F4/80+)/CD86+ macrophages increased significantly (P<0.01). The concentration of TGF-β decreased, while the concentrations of IL-1α and IL-6 increased significantly (all with P<0.01). Compared to that of the M1-polarized group, the relative expression of Gal-3 mRNA in the M1+Gal-3 inhibitor group was significantly decreased (P<0.01), the concentration of TGF-β significantly increased (P<0.01), while the IL-1α and IL-6 levels significantly decreased (both P<0.01). Compared to those of the M2-polarized group, the relative expression of Gal-3 mRNA in M2+Gal-3 inhibitor group was significantly decreased (P<0.05), along with decreased IL-1α and IL-6 and increased TGF-β concentrations (all with P<0.05). This study demonstrates that MMP-7 can activate Gal-3 expression to promote M2-type macrophage polarization and present anti-inflammatory activity.
The aim of this study is to explore the roles of cartilage interlayer protein 1 (CILP1) in the occurrence and development of colorectal cancer. The correlation of CILP1 expression and colorectal cancer stages, prognosis, and the Kirsten rat sarcoma viral oncogene (KRAS) pathway was analyzed by bioinformatics. The colorectal cancer cells SW620 and HCT116 were cultured and divided into 2 sets of groups (empty [Vector] group and CILP1 group [transfected with pcDNA 3.1-CILP1]; control group, KRAS inhibitor Salirasib [Salirasib] group, CILP1 group, and CILP1+Salirasib group). Cell proliferation, migration, invasion, and apoptosis were detected by CCK-8 assay, Transwell chamber assay, and FACS, respectively. The xenografted tumor model of colorectal cancer was established to observe the effects of CILP1 overexpression on tumor growth. The results showed that the expressions of CILP1 in patients with stages T3+T4, N+, and clinical stages Ⅲ+Ⅳwere higher than those with stages T1+T2, N0, and clinical stages Ⅰ+Ⅱ (P<0.05). The overall survival (OS) and the disease-free survival (DFS) in the low-expression CILP1 group were higher than those in the high-expression CILP1 group (P<0.05). Compared to those of the Vector group, the proliferation, migration, and invasion of tumor cells, volume and mass of tumors in the xenografted mouse model, and positive expression of CILP1 were significantly increased in the CILP1 group (P<0.05), while the apoptosis rate of tumor cells was significantly decreased (P<0.001). Gene set enrichment analysis (GSEA) showed that the KRAS pathway was significantly up-regulated in the high-expression CILP1 group (P<0.05). Compared to those of the control group, the expressions of CILP1, phosphorylated serine/threonine protein kinase 1 (p-Raf1), and phosphorylated mitogen-activated protein/extracellular signal-regulated kinase (p-MEK) were significantly increased in the CILP1 group (P<0.05), while significantly decreased in the Salirasib group (P<0.05). Compared to those of the CILP1 group, the expressions of CILP1, p-Raf1, and p-MEK were significantly decreased in the CILP1+Salirasib group (P<0.001). The above results suggest that CILP1 may promote the occurrence and development of colorectal cancer by activating KRAS pathway.
This study aims to investigate the protein and mRNA expressions of IL-22 and aryl hydrocarbon receptor (AhR)in the peripheral blood of patients with systemic lupus erythematosus (SLE), and to analyze the correlation between IL-22 and AhR and SLE disease activity index (SLEDAI). The expression levels of IL-22 and AhR protein in the peripheral blood of 43 SLE patients (25 in active stage and 18 in inactive stage) and 41 healthy controls were measured by ELISA. The expression levels of IL-22 and AhR mRNA in PBMC of SLE patients, healthy controls, and 31 rheumatoid arthritis (RA) patients by qRT-PCR. Spearman analysis was used to calculate the correlations of IL-22 and AhR protein and mRNA levels with the SLEDAI in SLE patients. ROC curve was used to analyze the combined diagnostic effect of anti-dsDNA antibody, IL-22, and AhR protein levels in SLE. The results showed that the levels of IL-22, AhR protein and mRNA were higher in SLE patients than those in healthy controls (P<0.001), and they were all higher in the active phase than in the inactive phase (P<0.05). IL-22 expressions were significantly elevated in SLE patients compared to RA patients (P<0.05). Spearman analysis showed that IL-22, AhR protein and mRNA levels were all positively correlated with SLEDAI in SLE patients (P<0.01). The AUC of combined diagnosis of IL-22, AhR, and anti-dsDNA antibody for SLE was 0.986, with a sensitivity and specificity of 90.7% and 97.6%, respectively. In conclusion, the protein and mRNA levels of IL-22, AhR in SLE patients are significantly higher than those in healthy subjects and are positively correlated with SLEDAI. The diagnosis of serum IL-22 and AhR combined with anti-dsDNA antibody has higher accuracy and may improve the clinical diagnosis of SLE in the future.
An improved ELISA method to detect cytokeratin 8 (K8) in the mouse intestinal mucosal injury model was evaluated in this study. The mouse intestinal mucosal injury models were established by either enteral injection of sodium formate and H2O2 or via partial ischemia caused by arteriolar clamps clipping of superior mesenteric artery branches. Pathological changes in intestinal tissue were observed and an optimized ELISA method was used to detect the level of K8 in stools within 24 h after surgery or in serum at different time points, and the change and expression difference of K8 were analyzed. Next, the ROC curve and the AUC were calculated to evaluate the authenticity of the detection method. The sensitivity and specificity were also analyzed to screen for the best cut-off value to provide a reference for clinical detection. The results showed that both enteral injection of sodium formate and H2O2 and arteriolar clamps clipping superior mesenteric artery branches caused significant pathological changes in intestinal tissue. The levels of K8 increased in stools within 24 h after surgery or in serum at different time points after ischemia and reperfusion. The improved ELISA method has high authenticity, and the optimal cut-off values for fecal and serum K8 are 69.78 μg/mL and 0.74 μg/mL respectively, with both high sensitivity and specificity. Utilizing this method to detect the level of K8 in feces or serum of mice reflected the degree of intestinal mucosal damage, therefore providing a reference for clinical monitoring of the occurrence and severity of intestinal mucosal injury, and has high application value.
This study aims to investigate the changes in mRNA expressions of stimulator of interferon gene (STING) and cyclic GMP AMP synthase (cGAS), the composition of myeloid dendritic cells (mDC) and their surface maturation markers CD80 and CD86, the related cytokines, and the correlation among above indicators to analyze the immune response upon adenovirus pneumonia infection. 34 children with acute stage adenovirus pneumonia and 34 healthy children were included in the study. The relative mRNA expression levels of STING and cGAS in the peripheral mononuclear cells were detected by qRT-PCR. The levels of IFN-α, IFN-β, TNF-α, and IL-6 were detected by ELISA. FACS was used to detect the mDC composition and surface markers CD80 and CD86. The correlation of the above indicators was analyzed. The results showed that compared to those of the control group, mRNA expression levels of STING and cGAS, IFN-α, IFN-β, TNF-α, IL-6, mDC cell number, CD80, and CD86 were all increased in adenovirus pneumonia patients (P<0.05). The level of mDC was positively correlated with the expressions of STING mRNA, cGAS mRNA, IFN-α, TNF-α, and IL-6. The CD80 level was positively correlated with STING mRNA, cGAS mRNA, IFN-α, IFN-β, TNF-α, and IL-6, whereas CD86 was positively correlated with IFN-α, IFN-β, and TNF-α (P<0.05). This study suggests that STING/cGAS genes are highly expressed in children with adenovirus pneumonia who exhibit increased mDC number, maturation markers, and cytokine secretion. The cGAS-STING pathway may be further activated to mediate innate immunity and exert antiviral effect.
This study aims to evaluate the immune-enhancing effect of compound fufangteng mixture (CFM) on B cells in cyclophosphamide-induced immunosuppression in BALB/c mice. BALB/c mice were randomly divided into normal control (NC) group, model control (MC) group, low-dose CFM (LD) group, medium-dose CFM (MD) group, high-dose CFM (HD) group, and positive control (PD) group. The expressions of CD19+B cell surface activating molecules (CD69, CD38) and exhaustion molecules (PD-1, CD95) in mice peripheral blood and spleen were detected by FACS. The results showed that compared to that of the NC group, the percentage of CD19+B cells in the peripheral blood of MC group decreased significantly(P<0.05), while the percentages of CD19+B cells in the peripheral blood of LD group, MD group, and HD group increased in a CFM dose-dependent manner. The expressions of CD69 and PD-1 of CD19+B cells in the peripheral blood of LD group and MD group were significantly higher than those of the NC group(P<0.05). There was no significant difference in the expression levels of CD69, CD38, PD-1, and CD95 of splenetic CD19+B cells among the six groups(P>0.05). Altogether, the results suggest that the CFM may improve the percentage of peripheral blood CD19+B cells and regulate CD69 and PD-1 in immunosuppression mice.
To investigate the correlation of rs767455 polymorphism at tumor necrosis factor receptor superfamily member 1A (TNFRSF1A) with spondyloarthritides (SpA) susceptibility and the clinical indicators, invasive probe based real-time PCR for rs767455 genotyping was employed in 112 SpA patients and 82 healthy controls. The statistical results showed that rs767455 was not significantly associated with the risk of SpA. However, using the combination of neutrophils and HLA-B27 to predict the occurrence of SpA increased the accuracy than HLA-B27 alone (AUC=0.972 of neutrophils+HLA-B27 vs AUC=0.944 of HLA-B27 alone, P=0.009). Correlation analysis using the mean or median of the clinical indicators for stratification showed that the risk of elevated platelet count in SpA patients with TC or CC genotype was significantly higher than that in TT genotype patients (TC+CC vs TT, OR=3.572, 95%CI 1.207-10.574, P=0.022). In addition, the risk of elevated platelet counts in TC genotype SpA patients was also higher than that in patients with TT genotype (TC vs TT, OR=3.907, 95%CI 1.195-12.778, P=0.024). Moreover, statistical analysis showed that compared to SpA patients carrying the T allele, patients carrying the C allele had a significantly higher probability of higher than average platelet count (C vs T, OR=3.000, 95%CI 1.143-7.871, P=0.026), monocyte count (C vs T, OR=2.794, 95%CI 1.110-7.033, P=0.029), and neutrophil count (C vs T, OR=2.794, 95%CI 1.110-7.033, P=0.029). Linkage analysis showed that rs767455 represented a splicing quantitative trait locus (sQTL) for TNFRSF1A across multiple tissues and cells. Altogether, the results suggest that neutrophils may participate in the pathogenesis of SpA independently of HLA-B27, and rs767455 T allele may play a role in the regulation of TNFRSF1A and lead to immune imbalance represented by elevated neutrophil levels in SpA patients. This study may guide in-depth research on both the pathogenesis and precise treatment of SpA.
In order to explore the mechanism of miR-29c-3p on the proliferation, migration, and invasion of cholangiocarcinoma cells by targeting urokinase-type plasminogen activator (PLAU), HuCCT1 and HCCC9810 cell lines with stable overexpression of miR-29c-3p were constructed and divided into miR-NC group, miR-29c-3p group, and miR-29c-3p+PLAU group. The differential expressions of miR-29c-3p and PLAU and their relationships with malignant clinical phenotypes of cholangiocarcinoma were analyzed by bioinformatics. The mRNA and protein expressions of miR-29c-3p and PLAU in each group were detected by RT-PCR and Western blotting, respectively. The proliferation, migration, and invasion of cells were detected by CCK-8 and Transwell assay, respectively. The target gene of miR-29c-3p was predicted by bioinformatics, and verified by dual luciferase reporter gene assay. The xenograft models of cholangiocarcinoma nude mice were constructed and divided into miR-NC group and miR-29c-3p group. The volume and mass of tumors were recorded. The results showed that the expression of miR-29c-3p was down-regulated in cholangiocarcinoma. Overexpression of miR-29c-3p inhibited the proliferation, migration, and invasion of cholangiocarcinoma cells, and the growth of xenografted tumor in nude mice (P<0.05). The expression of PLAU was up-regulated in cholangiocarcinoma, and abnormal expression of PLAU was correlated with malignant clinical phenotypes. Compared to that of the miR-NC group, the expression of miR-29c-3p was increased in miR-29c-3p group (P<0.05). Bioinformatics predicted that PLAU was the target gene of miR-29c-3p, which was verified by dual luciferase reporter gene assay. miR-29c-3p inhibited the proliferation, migration, and invasion of cholangiocarcinoma cells by targeting PLAU (P<0.05). This study suggests that miR-29c-3p can inhibit the proliferation, migration, and invasion of cholangiocarcinoma cells by negatively regulating the expression of PLAU.
To investigate the influences of naringin (NRG) on the proliferation and apoptosis of human rheumatoid arthritis fibroblast like synoviocytes (RA-FLS) mediated by the JAK/STAT3 pathway, RA-FLS were isolated from knee synovial tissue of RA patients and confirmed by the expressions of Vimentin and CD90 detected by immunofluorescence. RA-FLS were then divided into NC group (normally cultured RA-FLS), NRG-L group (20 μg/mL), NRG-M group (40 μg/mL), NRG-H group (80 μg/mL), and NRG-H+Coumermycin A1 (JAK activator) group (80 μg/mL NRG+10 μmol/L Coumermycin A1). The proliferation of RA-FLS was detected by the CCK-8 method and the morphology of apoptosis by Hoechst 33258 staining. The apoptotic rate of RA-FLS was detected by flow cytometry. The levels of IL-6, IL-1β and TNF-α in the culture supernatant of RA-FLS were detected by ELISA. The mRNA levels of proliferating cell nuclear antigen (PCNA) and Caspase-3 in RA-FLS were detected by qRT-PCR. And western blotting was used to detect the protein expressions of PCNA, Caspase-3, p-JAK2 and p-STAT3 in RA-FLS. The quality of RA-FLS isolation was confirmed by the spindle-shaped cells and strongly positive staining for vimentin and CD90. Compared to those of the NC group, the D(450 nm) value (24, 48 h) and the expressions of IL-6, IL-1β, TNF-α, PCNA at both mRNA and protein levels, as well as p-JAK2, p-STAT3 protein expressions were decreased of RA-FLS in the NRG-L group, NRG-M group and NRG-H group. On the other hand, the number of bright-staining cells, the apoptosis rate, and expression levels of Caspase-3 mRNA and protein were increased, in a dose-dependent manner (P<0.05). Compared to those of the NRG-H group, the D(450 nm) value (24 h, 48 h) and the levels of IL-6, IL-1β, TNF-α, PCNA at both mRNA and protein levels, as well as p-JAK2, p-STAT3 protein expression were increased in RA-FLS of NRG-H+Coumermycin A1 group, whereas the number of bright-staining cells, the apoptosis rate, Caspase-3 mRNA and protein expressions were decreased (P<0.05). Taken together, this study shows that NRG may inhibit the proliferation of RA-FLS and promote apoptosis by inhibiting JAK/STAT3 pathway.
Medulloblastoma (MB) is the most common pediatric brain malignancy, with a 5-year overall survival (OS) rate of around 70%. In recent years, the role of the tumor microenvironment on tumor growth has attracted extensive attention. The interaction between tumor cells and other cells in the tumor microenvironment including tumor-associated macrophages (TAM), T cells, NK cells, B cells, and tumor-associated astrocytes (TAA) plays an important role in the onset and progression of MB. Understanding the tumor immune microenvironment of this tumor, especially the relationship between tumor and immune cells, is crucial for the effective development of immune-based therapeutic strategies for MB. This review summarizes the latest research progress on the tumor microenvironment and immunotherapy of MB.
Natural killer cell receptor group Ⅱ member A (NKG2A) is an inhibitory receptor of the NKG2 receptor family, which is mainly expressed on a variety of immune cells represented by NK cells. Studies have also revealed that NKG2A is involved in the development of a variety of tumors. NKG2A targeting drugs are currently in clinical phase trials for multiple epithelial tumors. The review describes the biological properties of NKG2A and its regulatory role on CD8+ T cells, NK cells, and other immune cells. The review further explores the mechanism of NKG2A as an immune checkpoint in anti-tumor immunity, and provides a theoretical basis for clinical anti-tumor immunotherapy.
Inflammatory diseases are a series of acute or chronic diseases caused by inflammatory response to stimulation, which may seriously harm human health. Traditional Chinese medicine, which has multiple targets in immunomodulatory processes, plays a key role in the treatment of various inflammatory diseases. Corilagin is one of the major bioactive compounds in many medicinal plants. It has a variety of biological and pharmacological activities, including anti-oxidation, anti-inflammatory, liver protection, anti-virus, anti-hypertension, anti-infection, and anti-tumor. It has attracted extensive attention in recent years. However, there is a lack of systematic understanding of the role corilagin plays in inflammatory diseases. To better develop and utilize the medicinal plant resources in China, this review summarizes the role and related mechanism of corilagin in various inflammatory diseases. It will provide references for both follow-up basic research and clinical application of corilagin.
Allergic rhinitis and allergic asthma are the most common allergic airway diseases. IL-37 plays anti-inflammatory roles in allergic airway diseases by binding to IL-18Rα. On the other hand, IL-37 also binds to IL-18BP, which inhibits its binding to IL-18Rα. Therefore, a thorough understanding of the role of IL-37 in allergic airway diseases and its action mechanisms are imperative for the treatment of allergic airway diseases, and the development of IL-37-related biological agents.
Obesity increases the risk of infectious and neoplastic diseases. Studies have found that obesity may cause T cell metabolism disorders by affecting T cell lipid and carbohydrate metabolisms, promoting the release of inflammatory factors, and T cell aging, thereby increasing the mortality upon infection. Obesity is also an important risk factor associated with the prevalence of diseases. Obesity is associated with poor prognosis of neoplastic diseases, which is the combined effect of multiple factors affecting immune metabolism and inflammation. This review summarizes the research progress on T lymphocyte metabolism disorders caused by obesity and the role of T lymphocyte metabolism disorders in infectious and neoplastic diseases and treatment prospects.