Adiponectin alleviates LPS-induced inflammatory response of microglia through A20/NLRP3/caspase-1 pathway / 西安交通大学学报(医学版)

【Objective】 To explore the mechanism of adiponectin (APN) on lipopolysaccharide (LPS)-induced microglial inflammatory response. 【Methods】 After APN intervention or/and LPS stimulation, microglia were harvested and divided into control group, APN group, LPS group, and LPS+APN group. The expressions and secretion of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in microglia were detected by q-PCR and ELISA assay, respectively. The expressions of tumor necrosis factor α-induced protein 3 (TNFAIP3/A20), NOD-, LRR- and pyrin domain-containing 3 (NLRP3) and caspase-1 were detected by Western blot. After silencing A20 expression in the microglia, the cells were treated with APN, and the expressions of A20, NLRP3 and caspase-1 were detected by Western blot again. 【Results】 The mRNA transcription levels (P=0.018, P=0.009) and secretion (P=0.000 1, P=0.000 1) of TNF-α and IL-1β in LPS-induced microglia were significantly decreased after APN intervention. Moreover, the expression of A20 was increased while the expressions of NLRP3 and caspase-1 declined in microglia of LPS+ APN group when compared with LPS group (P=0.001, P=0.003, P=0.006). However, the down-regulated expressions of NLRP3 and caspase-1 by APN were reversed by A20 silencing in microglia (P=0.012, P=0.024). 【Conclusion】 APN can inhibit NLRP3 and caspase-1 expressions in microglia by the up-regulated expression of A20 protein.

Update in immune regulatory pathway of tumor necrosis factor-α-induced protein 8-like 2 / 中华危重病急救医学

The tumor necrosis factor-α-induced protein 8-like (TNFAIP8, TIPE) family is recently identified proteins consisting of four highly homologous mammalian proteins: TIPE, TIPE1, TIPE2, and TIPE3. Although the four members share similar molecular structure and function, involving effects in pathophysiological processes of inflammation, immunity, tumors, stroke, angiogenesis, and other diseases, they have individual characteristics. Many studies have shown that TIPE2 is an essential negative regulator of both innate and adaptive immunity. Up-regulation of TIPE2 expression can alleviate excessive inflammation during septic shock and maintain hemostasis of macrophages, neutrophils, dendritic cells, T cells, and B cells. In this review, we summarize the current literature for structure feature, immune function, and regulatory mechanism of TIPE2, together with its clinical significance in the pathogenesis of immune disorders of a wide array of human diseases. Understanding the basic biology of this new molecule might help us to seek novel strategies for the immunomodulation of human diseases.

The potential role and significance of tumor necrosis factor-α induced protein 8 like-2 in autoimmune diseases / 解放军医学杂志

Autoimmune disease is a condition arisen from an abnormal immune response to the tissue cells itself, its precise mechanism remains unknown, and the failure to distinguish self from non-self is often termed a breach of tolerance and is the basis for autoimmune illness. The tumor necrosis factor-α (TNF-α) induced protein 8 like-2 (TIPE2) is a newly discovered member of TNF-α induced protein 8 (TNFAIP8) family which is an essential negative controller of both innate and adaptive immunity. It has been documented that marked expressions of TIPE2 are evident in various autoimmune diseases, including autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), myasthenia gravis (MG) and systemic lupus erythematosus (SLE), which appear to be closely related to the severity, progression as well as prognosis of the illness, thereby contribute to the pathogenesis of autoimmune diseases. Deficient expression of TIPE2 might contribute to the hyper-reactivity of auto-reactive lymphocytes and macrophages, or aggregate inflammatory reaction by prompting high concentration of pro-inflammatory cytokines in peripheral blood, thus, trigger the development and progression of autoimmune diseases. In addition, dysregulation of immune homeostasis could be another latent target involved into the mechanism of autoimmune diseases. The present paper summarized the potential role and its mechanism of TIPE2 in the development of autoimmune diseases.

Expression of tumor necrosis factor -α-induced protein -8 -like 2 of bone marrow in children with aplastic anemia and its significance / 中华实用儿科临床杂志

Objective To investigate the characteristics of the bone marrow mesenchymal stem cell(MSC)in children with aplastic anemia(AA)in vitro,and the expressions of tumor necrosis factor -α-induced protein -8 -like 2(TIPE2)in the bone marrow,and the correlation between the level of TIPE2 mRNA with γ-interferon(IFN -γ)and IL -6 in AA patients.Methods Bone marrow samples were collected from 1 8 children with AA(AA group)and 8 children with bone injury (control group)who were hospitalized in Jinan Children′s Hospital from January 201 2 to June 201 5.MSC were isolated and cultured.The morphology of MSC was observed and immune phenotype was detected.The TIPE2 mRNA was detected by using real -time fluorescence quantitative PCR,and the levels of IFN -γand IL -6 were detected by using enzyme linked immunosorbent assay.Results Different sizes had been presented in the primi-tive MSC of AA patients,but the third passage MSC until 80% confluence had manifested the uniform convergence with long spindle and swirl distribution.In the sixth passage,cells showed degenerative change.The primitive and first pa-ssage MSC in patients with AA was longer than that in the controls.CD73 ,CD1 05 ,CD44 and CD90 were expressed in MSC,while CD34 ,CD45 ,CD271 expressed rarely.The level of TIPE2 mRNA in AA patients (5.29 ±1 .56)was obviously lower than that of the control group(8.68 ±2.00),and the difference was significant(t =-4.48,P 0.05).Conclusions The proliferation of MSC is significantly reduced in patients with AA.TIPE2,as an important role to stabilize the immune system,plays an important role in the occurrence of AA by its low expression and up -regula-ting the expression of inflammatory factors.

Study of tumor necrosis factor-αinduced protein 8 like-2 expression in peripheral blood mononuclear cells ;of patients with acute respiratory distress syndrome correlate with disease severity / 中华危重病急救医学

Objective To demonstrate the effect of tumor necrosis factor-α induced protein 8 like-2 (TIPE2) expression in patients with acute respiratory distress syndrome (ARDS) and its mechanism. Methods A prospective observation was conducted. Thirty-nine patients with ARDS admitted to department of emergency of PLA General Hospital from July 2013 to July 2015 were enrolled, and 35 healthy persons served as control group. The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score within 24 hours after admission, blood gas analysis, procalcitonin (PCT), and C-reactive protein (CRP) were recorded. The mRNA expressions of TIPE2 in peripheral blood mononuclear cell (PBMC) and myxoma resistance protein 1 (MX1) in plasma were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The correlations were analyzed by Spearman rank correlation analysis. Results The mean of APACHE Ⅱ score in 39 patients with ARDS was 25±3, the mean of PCT was (1.85±0.41) μg/L, and the mean of CRP was (18.0±3.0) mg/L. The TIPE2 mRNA expression in PBMC of ARDS patients was significantly down-regulated as compared with that of healthy control group (2-ΔΔCt: 3.28±0.15 vs. 8.87±0.20, P 0.05). The MX1 mRNA expression was positively correlated with APACHE Ⅱ score (r = 0.893, P 0.05). Conclusion TIPE2 expression was decreased in ARDS patients, which negatively correlate with disease severity, and indicate TIPE2 might be involved in the pathogenic process of ARDS.

Tumor necrosis factor-α induced protein 6 attenuates acute kidney injury following paraquat poisoning in rats / 中华危重病急救医学

Objective To explore the effects of tumor necrosis factor-α induced protein 6 (TSG-6) on acute kidney injury (AKI) following paraquat poisoning in rats.Methods Twenty-four male Sprague-Dawley (SD) rats were randomly divided into sham group (n=8),model group (n=8) and TSG-6-treated group (n=8) using a randomized number table.Rats were given an injection of 50 mg/kg of paraquat intraperitoneally (total volume was equalled to sterile normal saline) in model and TSG-6-treated groups.Rats in sham group were given 2 mg/kg of sterile saline.Mter 1 hour of paraquat administration,rats were treated with 30 μg of recombinant human TSG-6 intraperitoneally in TSG-6-treated group.After 6 hours of paraquat administration,serum was collected to assess renal function,then rats were sacrificed and renal tissues were immediately harvested.AKI score was evaluated by renal histopathology and gene expression of pro-inflammatory cytokines including interleukins (IL-1β and IL-6) and tumor necrosis factor-α (TNF-α) in kidney was assayed with real-time reverse transcription-polymerase chain reaction (RT-PCR).Results Compared with sham group,blood urea nitrogen (BUN),creatinine (Cr) and AKI score were significandy increased in model group [BUN (mmoUL):22.64 ±2.36 vs.7.09 ±0.65,t=6.986,P=0.000; Cr (μmol/L):177.28 ± 18.67 vs.60.32 ± 3.11,t=7.134,P=0.000; AKI score:9.14 ± 0.28 vs.0.30 ± 0.23,t=9.013,P=0.000].Moreover,the mRNA expressions of IL-1β,IL-6 and TNF-α were significantly elevated in model group (IL-1β mRNA:3.23 ± 0.28 vs.1.00 ±0.07,t=5.874,P=0.000; IL-6 mRNA:4.16 ±0.37 vs.1.00 ±0.08,t=7.125,P=0.000; TNF-α mRNA:3.85 ±0.31 vs.1.00 ±0.10,t=6.342,P=0.000).However,serum BUN,Cr,AKI score and the mRNA expressions of IL-1β,IL-6 and TNF-α in TSG-6-treated group were significantly lower than those in model group [BUN (mmol/L):14.07 ± 5.23 vs.22.64 ± 2.36,t=2.533,P=0.026; Cr (μmol/L):112.76 ± 14.81 vs.177.28 ± 18.67,t=2.778,P=0.016; AKI score:5.35 ±0.19 vs.9.14 ±0.28,t=2.885,P=0.013; IL-1β mRNA:2.26 ± 0.19 vs.3.23 ±0.28,t=2.457,P=0.023; IL-6 mRNA:2.92 ±0.29 vs.4.16 ±0.37,t=2.975,P=0.011; TNF-α mRNA:2.58 ± 0.23 vs.3.85 ± 0.31,t=2.564,P=0.019].Conclusion TSG-6 attenuates AKI following paraquat poisoning by suppressing inflammatory response.

Expression of tumor necrosis factor-α-induced protein 6 after transplantation of mesenchymal stem cells in a rat model of cardiopulmonary resuscitation / 中华急诊医学杂志

Objective To investigate the effects of bone marrow mesenchymal stem cells (MSCs)treatment on TSG-6 in a rat model of cardiopulmonary resuscitation (CPR).Methods Sprague Dawley (SD) rats were randomly (random number) divided into sham group,phosphate buffer solution (PBS)-treated group and MSCs-treated group.Animals were subjected to asphyxial cardiac arrest followed by CPR.In PBS-treated group or MSCs-treated group,animals were injected intravenously with PBS or MSCs at 2h after resuscitation.Neurological deficit scores (NDS) were assessed at 1,3 and 7 d after CPR.Serum S-100B was assayed using enzyme linked immunosorbent assay (ELISA).Immunofluorescence was performed to detect donor MSCs and the expression of TSG-6 in brain.TSG-6 and proinflammatory cytokines in brain were assayed using real time reverse transcription-polymerase chain reaction (RT-PCR).Western blot analysis was performed to measure the levels of neutrophil elastase (NE) in brain.Multiple comparisons were made by analysis of variance.Results At 3d and 7d,MSCs-treated group demonstrated higher NDS than PBS-treated group (P ＜ 0.01),and serum S-100B levels significantly reduced in MSCs-treated group compared with PBS-treated group (P ＜ 0.01).DAPI-labeled MSCs migrated into the ischemic brain and some DAPI + cells colocalized with TSG-6.Compared with PBS-treated group,MSCs treatment significantly up-regulated the expression of TSG-6 and reduced the expression of NE and proinflammatory cytokines in brain at 3 d and 7 d after CPR (P ＜ 0.05).Conclusion Systemically administered MSCs suppressed inflammatory responses in brain after CPR and improved neurological function in rats possibly via induction of TSG-6.