Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

Objective: To investigate the effect of microRNA (miR-148b) targeting decoy receptor 3 (DcR3) on macrophage polarization in sepsis. Methods: Experimental study. From December 2019 to December 2022, serum microRNA expression was detected in 3 patients with sepsis and 3 healthy controls in the clinical laboratory of Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Phorbol 12-myristate 13-acetate (PMA) was used to induce the differentiation of human acute monocytic leukemia cells THP-1 into macrophages, and then lipopolysaccharide (LPS) was added to stimulate the establishment of a sepsis cell model, and the expression changes of miR-148b and DcR3 were detected by RT-PCR and Western blot. Overexpression of DcR3 was used to detect the expression levels of TNF-α, CD163 and IL-10 in macrophages stimulated by LPS (100 ng/ml). Overexpression of miR-148b was used to observe the changes of molecular markers of macrophage polarization. The targeting regulation effect of miR-148b on DcR3 was determined by dual-luciferase reporter assay. t test was used to analyze whether there were statistical differences among the groups. Results: The expression of miR-148b was down-regulated (P<0.05) and the expression of DcR3 was up-regulated (P<0.01) in THP-1 macrophages stimulated by LPS. Overexpression of DcR3 inhibited the expression of TNF-α (P<0.05) and promoted the expression of CD163 (P<0.01) and IL-10 (P<0.01). When miR-148b mimics was added, the opposite effect was observed. The dual-luciferase reporter assay confirmed that miR-148b targets and binds to DcR3, inhibiting its transcription and expression. The results of flow cytometry showed that DcR3 could reverse the promoting effect of miR-148b on the CD86/CD163 ratio of macrophages (P<0.05). Conclusion: miR-148b inhibits the expression of DcR3, thereby inhibiting M2 polarization in LPS-stimulated macrophage cells.
Humans ; Interleukin-10 ; Lipopolysaccharides/pharmacology* ; Macrophages ; MicroRNAs/genetics* ; Receptors, Tumor Necrosis Factor, Member 6b/metabolism* ; Tumor Necrosis Factor-alpha

No abstract available.
Female ; Gastric Mucosa/*metabolism ; Helicobacter Infections/*genetics ; Humans ; Male ; Peptic Ulcer/*genetics ; *Promoter Regions, Genetic ; Tumor Necrosis Factor-alpha/*genetics

Animals ; Lung ; drug effects ; metabolism ; Perchlorates ; toxicity ; Quaternary Ammonium Compounds ; toxicity ; RNA, Messenger ; genetics ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Astrocytes/metabolism* ; Cell Hypoxia ; Cells, Cultured ; Cytokines ; Humans ; Lipopolysaccharides ; Membrane Proteins/genetics* ; Proto-Oncogene Proteins/genetics* ; Tumor Necrosis Factor-alpha

The onset of inflammatory bowel disease (IBD) involves many factors, including environmental parameters, microorganisms, and the immune system. Although research on IBD continues to expand, the specific pathogenesis mechanism is still unclear. Protein modification refers to chemical modification after protein biosynthesis, also known as post-translational modification (PTM), which causes changes in the properties and functions of proteins. Since proteins can be modified in different ways, such as acetylation, methylation, and phosphorylation, the functions of proteins in different modified states will also be different. Transitions between different states of protein or changes in modification sites can regulate protein properties and functions. Such modifications like neddylation, sumoylation, glycosylation, and acetylation can activate or inhibit various signaling pathways (e.g., nuclear factor-‍κB (NF-‍κB), extracellular signal-regulated kinase (ERK), and protein kinase B (AKT)) by changing the intestinal flora, regulating immune cells, modulating the release of cytokines such as interleukin-1β (IL-‍‍1β), tumor necrosis factor-α (TNF‍-‍α), and interferon-‍γ (IFN-‍γ), and ultimately leading to the maintenance of the stability of the intestinal epithelial barrier. In this review, we focus on the current understanding of PTM and describe its regulatory role in the pathogenesis of IBD.
Cytokines/genetics* ; Humans ; Inflammatory Bowel Diseases ; NF-kappa B/metabolism* ; Protein Processing, Post-Translational ; Tumor Necrosis Factor-alpha/metabolism*

OBJECTIVETo explore the molecular biology mechanism of acupuncture on improving immune function damage induced by chemotherapy in cancer-bearing mice.

METHODSSeventy-two mice (36 mice in 3-day treatment and 5-day treatment, respectively) which were successfully made into cancer-bearing model were divided into a blank group, a model group, an acupuncture group and a moxibustion group by stratified randomization method, 9 mice in each one. Except for the mice in the blank group, the remaining mice were treated with intra-peritoneal injection of cyclophosphamide (CTX, 150 mg/kg), to establish the cancer-bearing mice of CTX. The mice in the blank group were treated with intraperitoneal injection of 0.9% NaCl (identical dose as other groups). After 4 h, the mice in the acupuncture group and moxibustion group were treated with acupuncture and moxibustion at "Dazhui" (GV 14), "Geshu" (BL 17), "Shenshu" (BL 23), "Zusanli" (ST 36), once a day. The mice in the blank group and model group were treated with immobilization and fixation during the same time. On the next day of the end of 3-day and 5-day treatment, the sample was collected. The ELISA method was applied to measure the contents of IL-12 and TNF-&alpha; in spleen of all the mice.

RESULTSAfter 3-day and 5-day treatment, compared with the blank group, the contents of IL-12 and TNF-&alpha; in spleen in the model group were reduced (all P < 0.05); compared with the model group, the contents of IL-12 and TNF-&alpha; in spleen in the acupuncture group and moxibustion group were increased (all P < 0.05), but the content of IL-12 and TNF-&alpha; in the acupuncture group was not different from that in the moxibustion group (both P > 0.05).

CONCLUSIONAcupuncture and moxibustion could effectively increase the contents of IL-12 and TNF-&alpha; in spleen of CTX cancer-bearing mice, which could relieve chemotherapy-induced immune function damage to improve immune function.

Acupuncture Therapy ; Animals ; Cyclophosphamide ; adverse effects ; Humans ; Interleukin-12 ; genetics ; metabolism ; Male ; Mice ; Moxibustion ; Neoplasms ; chemically induced ; genetics ; metabolism ; therapy ; Spleen ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

A main symptom of equine metabolic syndrome (EMS) in ponies is pathological obesity characterized by abnormal accumulation of fat deposits and inflammation. In this study, we analyzed the expression of two pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), in subcutaneous adipose tissue and the correlation with serum concentrations in peripheral blood of Welsh ponies. Based on clinical examination findings, the animals were divided into two groups: ponies affected with EMS (n = 8) and obese ponies (n = 8). The adipose tissue was examined using immunohistochemical analysis while concentrations IL-6 and TNF-alpha were measured using enzyme-linked immunosorbent assays (ELISAs). Additionally, histological characterization of the adipose tissue was performed. The results obtained showed that IL-6 expression in adipose tissue biopsies derived from animals with EMS was enhanced while TNF-alpha levels of both groups were comparable. Compared to the obese ponies, EMS animals also had significantly elevated levels of serum IL-6 and TNF-alpha. Histological analysis revealed macrophage infiltration and fibrosis in adipose tissue preparations from the EMS group. These data suggest that IL-6 may play a key role in the course of EMS in Welsh ponies. Our findings also demonstrated that analysis of pro-inflammatory cytokines levels in serum may serve as an additional tool for diagnosing EMS.
Adipose Tissue/*metabolism ; Animals ; Female ; Horse Diseases/blood/*metabolism ; Horses ; Interleukin-6/blood/genetics/*metabolism ; Male ; Metabolic Syndrome X/metabolism/*veterinary ; Tumor Necrosis Factor-alpha/blood/genetics/*metabolism

OBJECTIVE: To study the role of Novaferon on TNF-α production and expression of NF-κB mRNA in monocytes isolated from normal human peripheral blood and to provide theoretical basis for treatment of immunological diseases with Novaferon. METHODS: Monocytes were isolated from the peripheral blood in 30 healthy volunteers and divided into 5 groups: group A was blank control, group B was stimulated by LPS without Novaferon intervention, group C by LPS together with Novaferon intervention, group D by LPS before Novaferon intervention, which group E by LPS after Novaferon intervention. We detected the concentration of TNF-α after LPS stimulation and Novaferon intervention in the supernatant by ELISA and expression of NF-κB mRNA by RT-PCR. RESULTS: Novaferon inhibited TNF-α production by monocytes isolated from healthy volunteers induced by LPS in vitro in group D compared with group B [(1446.76±72.36) pg/mL vs (946.46±46.12) pg/mL, P<0.01], and the rate was 29.7%. There was no significant change in TNF-α concentration in group C and E compared with group B [(1446.76±72.36) pg/mL vs (1275.62±87.75) pg/mL, P>0.05; (1446.76±72.36) pg/mL vs (1383.62±86.96) pg/mL, P>0.05]. There was significant change in NF-κB mRNA expression in group D compared with group B (0.2829±0.0365 vs 0.4994±0.0604, P<0.01). There was no significant change in NF-κB mRNA expression in group C and group E compared with group B (0.4716±0.0616 vs 0.4994±0.0604, P>0.05; 0.4767±0.0600 vs 0.4994±0.0604, P>0.05). CONCLUSION Novaferon can suppress TNF-α secretion by monocytes induced by LPS in vitro, and it can affect the immunity function of monocytes, which may be associated with the downregulation of NF-κB mRNA expression in monocytes.
Humans ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; metabolism ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; metabolism

BACKGROUNDAlthough the onset of anemia during infectious disease is commonly correlated with production of inflammatory cytokines, the mechanisms by which cytokines induce anemia are poorly defined. This study focused on the mechanism research.

METHODSDifferent types of mice were infected perorally with Toxoplasma gondii strain ME49. At the indicated times, samples from each mouse were harvested, processed, and analyzed individually. Blood samples were analyzed using a Coulter Counter and red blood cell (RBC) survival was measured by biotinylation. Levels of tumor necrosis factor-&alpha; (TNF-&alpha;), inducible nitric oxide synthase (iNOS), and inducible protein 10 (IP-10) mRNA in liver tissue were measured by real-time polymerase chain reaction.

RESULTST. gondii-infected mice exhibited anemia due to a decrease in both erythropoiesis and survival time of RBC in the circulation (P < 0.02). In addition, infection-stimulated anemia was associated with fecal occult, supporting previous literature that hemorrhage is a consequence of T. gondii infection in mice. Infection-induced anemia was abolished in interferon gamma (IFNγ) and IFNγ receptor deficient mice (P < 0.05) but was still evident in mice lacking TNF-&alpha;, iNOS, phagocyte NADPH oxidase or IP-10 (P < 0.02). Neither signal transducer and activator of transcription 1 (STAT1) deficient mice nor 129S6 controls exhibited decreased erythropoiesis, but rather suffered from an anemia resulting solely from increased loss of circulating RBC.

CONCLUSIONSInfection-stimulated decrease in erythropoiesis and losses of RBC have distinct mechanistic bases. These results show that during T. gondii infection, IFNγ is responsible for an anemia that results from both a decrease in erythropoiesis and a STAT1 independent loss of circulating RBC.

Anemia ; genetics ; metabolism ; Animals ; Erythrocytes ; pathology ; Interferon-gamma ; metabolism ; Male ; Mice ; Mice, Knockout ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Receptors, Interferon ; genetics ; metabolism ; STAT1 Transcription Factor ; genetics ; metabolism ; Toxoplasma ; pathogenicity ; Tumor Necrosis Factor-alpha ; genetics ; metabolism