Expression level and clinical significance of SOCS1 in the serum of patients with chronic HBV-related liver disease / 西安交通大学学报(医学版)

【Objective】 To study the expression levels of suppressor of cytokine signaling 1 (SOCS1) and its clinical significance in hepatitis B virus (HBV)-related liver diseases. 【Methods】 For this study we enrolled 25 patients with chronic hepatitis B (CHB), hepatitis B cirrhosis, or HBV-associated chronic acute liver failure (HBV-ACLF), and 25 healthy controls. The expression levels of SOCS1 mRNA in peripheral blood mononuclear cells (PBMCs) were determined using the RT-PCR method. The levels of SOCS1 and interleukin-6 (IL-6) in the plasma of patients with chronic liver diseases and healthy controls were measured using the ELISA method. The relative expression levels of SOCS1, SOCS1 mRNA, and other laboratory test indicators such as HBV-DNA, alanine aminotransferase (ALT), aspartate aminotransferase (AST), prothrombin activity (PTA) and total bilirubin (TBil) were compared among the groups. Additionally, the correlation between the expression levels of SOCS1 mRNA and the aforementioned laboratory indicators was assessed. 【Results】 The expression levels of SOCS1 mRNA and serum SOCS1 were highest in the HBV-ACLF group, followed by the cirrhosis group, and lowest in the healthy control group, with statistically significant differences (F=109.65, P<0.001). The relative expression of SOCS1 mRNA was positively correlated with TBil (r=0.89, P<0.001), ALT (r=0.89, P<0.001), AST (r=0.84, P<0.001) and IL-6 (r=0.93, P<0.001), but negatively correlated with PTA (r=-0.89, P<0.001) and was not significantly correlated with HBV-DNA (P=0.28). 【Conclusion】 The expression levels of SOCS1 in patients with HBV-related chronic liver diseases can reflect the severity of the disease and show a significant correlation with indicators used to assess the severity of liver diseases.

Correlation between serum miR-155, SOCS-1 levels and renal interstitial fibrosis in patients with IgA nephropathy / 中国医师杂志

Objective:To investigate the changes of serum micro ribonucleic acid-155 (miR-155) and suppressor of cytokine signaling-1 (SOCS-1) levels in patients with IgA nephropathy and their relationship with renal interstitial fibrosis.Methods:A total of 365 patients with primary IgA nephropathy admitted to Jining First People′s Hospital from January 2009 to June 2018 were selected as the research objects. According to the degree of renal interstitial fibrosis, the patients were divided into T0 group (139 cases), T1 group (124 cases) and T2 group (102 cases). In addition, 361 healthy subjects who had physical examination in our hospital in the same period were selected as the healthy control group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-155 in serum of all the subjects; the levels of serum SOCS-1, transforming growth factor-β1 (TGF-β1) and monocyte chemoattractant protein-1 (MCP-1) were detected by enzyme-linked immunosorbent assay (ELISA). Logistic regression model was used to analyze the influencing factors of renal interstitial fibrosis in patients with IgA nephropathy; Pearson method was used to analyze the correlation between serum miR-155, SOCS-1 levels and influencing factors of renal interstitial fibrosis, TGF-β1 and MCP-1; receiver operating characteristic (ROC) curve was used to analyze the predictive value of serum miR-155 and SOCS-1 levels in patients with IgA nephropathy.Results:The levels of systolic blood pressure (SBP), diastolic blood pressure (DBP), 24 h urine protein, serum creatinine, serum uric acid, miR-155, TGF-β1, MCP-1, urinary retinol binding protein (RBP) and acetyl β D-glucosaminidase (NAG) in healthy control group, T0 group, T1 group and T2 group were significantly increased in turn, while the levels of hemoglobin, estimated glomerular filtration rate (eGFR), urinary osmotic pressure and serum SOCS-1 were significantly decreased in turn (all P<0.05). High SBP, high DBP, low hemoglobin, high serum creatinine, high uric acid, high 24-hour urine protein and low eGFR level were independent risk factors of renal interstitial fibrosis in patients with IgA nephropathy (all P<0.05). The serum miR-155 level was positively correlated with TGF-β1, MCP-1, SBP, DBP, serum creatinine, serum uric acid levels and 24 h urine protein, but negatively correlated with SOCS-1, hemoglobin and eGFR levels (all P<0.05). The serum SOCS-1 level was negatively correlated with TGF-β1, MCP-1, SBP, DBP, serum creatinine, uric acid levels and 24 h urine protein, but positively correlated with hemoglobin and eGFR levels (all P<0.05). The area under the curve of predicting the occurrence of renal interstitial fibrosis in patients with IgA nephropathy by serum miR-155 and SOCS-1 combined detection was 0.882, which was significantly larger than that by serum miR-155 and SOCS-1 alone ( P<0.05). Conclusions:The expression of miR-155 is up-regulated and SOCS-1 is down-regulated in IgA nephropathy patients, they may be used as predictors to evaluate the occurrence of renal interstitial fibrosis.

Research progress on the role of miR-155 in rejection of organ transplantation / 器官移植

Rejection has constantly been an unresolved challenge in the field of organ transplantation. The research on the mechanism of rejection plays a significant role in improving the efficacy of organ transplantation and enhancing the survival rate of graft. The innate and specific immune responses of the human body jointly participate in the graft rejection, leading to graft injury. In recent years, multiple researchers have conducted in-depth studies on the mechanism underlying the role of microRNA (miR) in regulating rejection. Among them, miR-155 has been widely considered as a key factor involved in immune regulation. The expression level and functional status of miR-155 may be intimately associated with the occurrence of rejection, which may become a new target for overcoming rejection. In this article, relevant studies on the role of miR-155 in regulating key immune cells in innate and specific immune responses were reviewed, aiming to provide novel ideas for the development of new immunosuppressants and rejection therapy.

Mechanism of silencing miR-4320 expression in inhibiting proliferation and migration of gastric cancer cells / 国际外科学杂志

Objective:To explore the effect and mechanism of microRNA (miRNA)-4320 on the proliferation and migration of gastric cancer MGC803 cells.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-4320 in four gastric cancer cell lines(MGC803, HS-746T, SGC7901, BGC823) MGC803 cells were infected with recombinant lentivirus carrying miR-4320 interference fragments or blank lentivirus, and set as si-miR-4320 group and NC group. Thiazole blue colorimetry and Transwell small box experiment were used to detect the proliferation and migration of MGC803 cells after miR-4320 was down-regulated. The bioinformatics software RNAhybrid was used to predict the target gene of miR-4320. The targeting relationship between miR-4320 and target gene was verified by dual-luciferase reporter gene experiment. qRT-PCR and Western blot were used to detect the expression of miR-4320 target gene. Measurement data were expressed as mean ± standard deviation ( ± s), and t-test or one-way ANOVA was used for comparison between groups. Results:The expression of miR-4320 in the four gastric cancer cell lines was significantly higher than that of normal gastric mucosal epithelial cells ( P<0.01). The expression of miR-4320 in MGC803 cells in the NC group and the si-miR-4320 group were 8.19±1.00 and 1.09±0.31, respectively. The miR-4320 interference fragment significantly reduced the expression of miR-4320 ( P<0.01). The absorbance of MGC803 cells in the si-miR-4320 group was significantly lower than that of the NC group ( P<0.05), and the migration ability was significantly lower than that of the NC group ( P<0.01). Suppressor of cytokine signaling1 ( SOCSI) is the target gene of miR-4320. Compared with the NC group, the SOCS1 gene expression in the si-miR-4320 group was significantly up-regulated ( P<0.01). Conclusions:The expression of miR-4320 is increased in gastric cancer cell lines. Down-regulating the expression of miR-4320 can inhibit the proliferation and migration of gastric cancer MGC803 cells by inducing the expression of SOCS1 gene.

Mechanism of action of suppressor of cytokine signaling 1 in the development and progression of liver inflammatory diseases / 临床肝胆病杂志

The development of liver inflammatory diseases is associated with autoimmunity and inflammatory response. As a negative feedback regulator of cell signal, suppressor of cytokine signaling 1 (SOCS1) plays a key role in the development and progression of inflammatory diseases. This article mainly introduces the mechanism of action of SOCS1 in autoimmunity and inflammatory response and briefly describes its role in the development and progression of liver inflammatory diseases such as viral hepatitis and nonalcoholic steatohepatitis. The analysis shows that the abnormal expression of SOCS1 in inflammatory response is associated with the regulation of cytokine receptor, Toll-like receptor, and hormone receptor signal, which leads to the development of inflammatory diseases. Therefore, SOCS1 has potential prospects as an auxiliary means for the diagnosis and treatment of liver inflammatory diseases.

Opposite effects of miR-155 in the initial and later stages of lipopolysaccharide (LPS)-induced inflammatory response / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Although microRNA-155 (miR-155) is considered a pro-inflammatory mediator, cumulative evidence indicates that it also has anti-inflammatory effects in macrophages and dendritic cells. In this study, we identified the dramatic expression changes of more than half of potential miR-155-targeted genes upon lipopolysaccharide (LPS) stimulation; 223 genes were down-regulated and 85 genes were up-regulated, including suppressor of cytokine signaling 1 (

Expression of SOCS1 in overload papillary muscles of mice cardiac ventricle / 国际生物医学工程杂志

Objective Toinvestigatetheexpressionofsuppressorofcytokinesignaling1(SOCS1)inoverloaded ventricle papillary muscle, so as to understand its expression characteristics in structural remodeling after the overloading and the biomechanical properties of the muscle under cubic jellyfish toxin-1(CfTX-1) pretreatment that can affect cell signal transduction. Methods Abdominal aortic-venous fistula (AVF) were operated in Kunming mice (n=5), and the cardiac left ventricles were harvested after two weeks of fistulation. The mice in normal group were sham operated as a control (n=5). In vitro culture, the left ventricular papillary muscle of normal mice was used (n=20). In the stretching group, the isolated papillary muscles were double-ratio stretched and fixed on silicone plate. In the relaxation group, the muscles were not stretched. A separated subgroup that transfected with SOCS1 plasmids were set in each group of stretching and relaxation. The papillary muscle samples of each group were cultured in culture medium for 3 days at 37 ℃, and then homogenized for extracting total protein. The total protein was separated by 10％ sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The 23 ku band with SOCS1 was used as the target band, and the integrated optical density (IOD) value was measured by computer image analysis method. The expression of SOCS1 protein was detected by Western Blot and the imprinted IOD value was also measured. The papillary muscle in the stretching group was stretched by micro-positioned stretching method, and the initial load was 1 g. After stabilization, the papillary muscle was stretched by 15 mm for continuously 5 times, and the passive tension characteristic curves during the first and fifth stretching were observed and recorded. The peak passive tension (PTmax) and its deceleration velocity (DV) of the papillary muscle were calculated based on the curves. Results Comparing with the AVF group, the normal group had higher IOD values of 23 ku band and SOCS1 blot in total protein of the papillary muscle, and the differences were statistically significant (all P<0.01). The IOD value of 23 ku band in the SOCS1 transfected stretching group was significantly higher than those of the two relaxation groups, and the differences were statistically significant (all P<0.01). However, the difference of this value was not statistically significant between the two relaxation groups. The average IOD value of SOCS1 blot in the SOCS1 transfected stretching group was higher than those of the normal stretching group and the SOCS1 transfected relaxation group, and the differences were statistically significant (all P<0.01). Comparing with the normal group, the AVF group had higher PTmax and ultimate PTmax of the papillary muscles, and had a lower DV values, and the differences were statistically significant (all P<0.01). Conclusions The expression of SOCS1 is sensitive to tension load, and has a positive effect as an overload-sensitive signal in improving myocardial adaptability, protecting myocardial structure and maintaining systolic and diastolic function. CfTX-1 also has a positive effect on improving the compliance of ventricular papillary muscles.

Carboxymethytl pachymaram induces the methylation of SOCS-1 gene and the maturation of human monocyte-derived dendritic cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of carboxymethytl pachymaram ( CMP ) on the methylation of SOCS-1 (suppressor of cytokine signaling-1) gene and the in vitro maturation of human mono-cyte-derived dendritic cells (DCs).Methods Human DCs were induced from the peripheral blood mono-cytes in vitro with the treatment of recombined human GM-CSF and interleukin-4 ( IL-4 ) and cultured with different concentrations of CMP (10, 50, and 100 mg/L).The methylation and expression of SOCS-1 gene were analyzed by methylation-specific polymerase chain reaction (MSP) and real-time PCR, respectively. The phenotypic markers of DCs were detected by flow cytometry .Mixed lymphocyte reaction ( MLR) and ELISA were performed to measure the lymphocyte proliferation induced by DCs and IL-12 secretion by DCs . Results CMP promoted the methylation of SOCS-1 gene, but inhibited the expression of SOCS-1 gene in dendritic cells at the concentrations of 50 mg/L and 100 mg/L.The expression of phenotypic markers (CD80, CD83, CD86 and HLA-DR), IL-12 secretion and lymphocyte proliferation induced by DCs were significantly enhanced in a dose dependent manner with the treatment of CMP .Compared with control group , the levels of methylated SOCS-1 gene and IL-12 and the lymphocyte proliferation index were increased upon the stimulation with 50 mg/L and 100 mg/L of CMP (P<0.01), but the expression of SOCS-1 gene was de-creased.The expression of CD80, CD83 and HLA-DR on DCs in the presence of 100 mg/L of CMP were higher than those of control group (P<0.05).Conclusion CMP could induce the methylation of SOCS-1 gene and the maturation of DCs derived from peripheral blood monocytes .

Effect of miRNA-155 on phenotype and function of regulatory T cell / 器官移植

Objective Toinvestigatetheeffectofmicroribonucleicacid(miRNA)-155ontwo subtypesofregulatoryTcell(Treg):inducedTreg(iTreg)andnaturalTreg(nTreg).Methods NaveT cells and nTreg were isolated from peripheral blood mononuclear cell (PBMC)of healthy donors by magnetic cell sorting. Cells were divided into 3 groups during culture,including control group (nave T cells were cultured with the presence of interleukin-2 ),iTreg group (nave T cells were cultured with the presence of interleukin-2 and transforming growth factor-β)and nTreg group(nTreg cells was cultured with interleukin-2).Each group was divided into 3 subgroups (none,scramble or miRNA-155 antagomir subgroup,3 wells in each subgroup). Expression level of miRNA-155 gene of none subgroup in 3 groups was detected by low density chip analysis method. The levels of surface marker CD25,Foxp3,CD127 of each subgroup in 3 groups were detected by flow cytometry. The percentage of CD4 +CD25 +Foxp3 +SOCS1 +Treg and suppressive function of Tregofeachsubgroupin3groupswerealsodetectedbyflowcytometry.Results Comparedwithcontrolgroup and iTreg group,the expression level of miRNA-155 was significantly lower and SOCS1 was significantly higher in nTreg group (all in P<0.05 ). After the addition of miRNA-155 antagomir,no significant change was observed in the important surface markers of Treg like Foxp3,CD25,CD127. Compared with control group and iTreg group,the expression of SOCS1 in nTreg group increased significantly (both in P <0.05 ). The expression level of miRNA-155 of none subgroup in iTreg group was lower. The expression of SOCS1 increased after the miRNA-155 was inhibited by antagomir (miRNA-155 antagomir subgroup). In iTreg group,the suppressive function of Treg in miRNA-155 antagomir subgroup was higher than that in none subgroup at the ratioof1∶8,1∶16and1∶32(allinP<0.05).Conclusions AntagonismofmiRNA-155invitrohasno significant effect on the suppression function of nTreg,but can increase the SOCS1 expression level and suppression in vitro of iTreg.

Influence of SOCS-1 on AGEs-induced renal tubular epithelial-myofibroblast transdifferentiation / 中国药理学通报

Aim To investigate the effects of suppressor of cytokine signaling-1(SOCS-1)on advanced glycation end products(AGEs)induced-renal tubular epithelial-myofibroblast transdifferentiation and activation of JAK/STAT in cultured human renal tubular epithelial cells(HKC).Methods Stable transfections of HKC with pCR3.1 vector and pCR3.1/SOCS-1 were performed with Lipofectamine 2000,and cells were selected with geneticin.Cells were stimulated with BSA and AGEs. The protein expressions of SOCS-1,α-SMA,E-cadherin,Col I,signal transducer and activatior of transcription 1,3(STAT1,STAT3),p-STAT1 and p-STAT3 were observed by Western blot.The protein synthesis of TGF-β_1 in the supernatants of the HKC was detected by enzyme-linked immunoadsorbent assay(ELISA).α-SMA and E-cadherin mRNA were measured by reverse transcription and polymerase chain reaction(RT-PCR).Results Compared with control group,the expression levels of α-SMA protein and mRNA and Col I were significantly increased in HKC with AGEs stimulation and there was a higher concentrations of TGF-β_1 in the supernatants.However,the expression of E-cadherin protein and mRNA were decreased with AGEs stimulation.Overexpression of SOCS-1 inhibited AGEs-induced activation of STAT1 and STAT3 and high expression of α-SMA protein and mRNA and Col I,and reversed the expression of E-cadherin protein and mRNA induced by AGEs.Meanwhile,overexpression of SOCS-1 reduced the concentration of TGF-β_1 in the supernatants of HKC with AGEs stimulation.Conclusion Overexpression of SOCS-1 inhibits AGEs-induced renal tubular epithelial-myofibroblast transdifferentiation maybe partly through blocking activation of JAK/STAT.

Expressions of SOCS-1 and SOCS-3 in the myocardium of patients with sudden cardiac death / 世界急诊医学杂志（英文）

BACKGROUND:As the regulators of cytokines, suppressors of cytokine signaling (SOCS) play an important role in the inflammation reaction. Some studies found that SOCS-1 and SOCS-3 were involved in the pathogenesis of some inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease. But the expressions of SOCS in coronary heart disease have not yet been reported. This study aimed to investigate the expression and clinical significance of SOCS-1 and SOCS-3 in the myocardium of patients with sudden cardiac death (SCD).METHODS:Myocardial autopsy specimens were collected from 24 patients at the Forensic Medicine Department of Sun Yat-Sen University, Guangzhou, China between 2005 and 2006. Of them, 9 patients had autopsy findings consistent with coronary atherosclerosis (non-myocardial infarction) leading to SCD (non-MI group), 7 died of acute myocardial infaction (MI group), and 8 died from traffic accidents and trauma (control group). The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the myocardium of the non-MI, MI and control groups were detected using RT-PCR. The levels of SOCS-1 and SOCS-3 proteins were detected using immunohistochemistry. Statistical analyses were performed using SPSS version 13.0 software and the data were analyzed by ANOVA.RESULTS:The expressions of SOCS-1 mRNA and SOCS-3 mRNA in the non-MI and MI groups were significantly higher than those in the control group[(0.788±0.101), (0.741±0.111) vs. (0.436±0.044), (P<0.01); (0.841±0.092), (0.776±0.070) vs. (0.454±0.076), (P<0.01)] respectively. The antibody-positive cells of SOCS-1 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(320.00±48.48), (347.14±70.88) vs. (42.50±10.35), (P<0.01)] respectively. The antibody-positive cells of SOCS-3 protein in the myocardium of the non-MI and MI groups were significantly higher than those in the myocardium of the control group[(381.11±59.25) vs. (40.00±10.69), (P<0.01)] and[(332.86±111.91) vs. (40.00±10.69), (P=0.001)].CONCLUSION:The expressions of SOCS-1 and SOCS-3 in the myocardium of patients with SCD from coronary heart disease are significantly increased and contribute to the pathogenesis of SCD.

The expression of STATI/SOCS1 on the rat synoviocytes induced by high mobility group box / 中华风湿病学杂志

Objective To investigate the effect and possible mechanisms of high mobility group box (HMGB) 1 on the proliferation of RSC-364 synoviocytes. Methods ① RSC-364 cells stimulated by 10 μg/L TNF-α and cells of the normal control groups were collected at 6, 12, 24 h respectively in vitro. HMGB1mRNA and protein was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (ICC); ②RSC-364 cells induced by 10 μg/L HMGB1 were collected in 6, 12, 24 h respectively, so did normal control group cells in vitro. The expression of signal transducer and activator of transcription (STAT) mRNA 1 was detected by RT-PCR. The expression of STATland SOCSI proteins were detected by ICC and flow cytometry analysis (FCM). The expression of PCNA was detected by ICC. Results ① Compared with the control group, TNF-α markedly up-regulated HMGBI mRNA at 6, 12, 24 h respectively [0.86, 0.92, 1.06 vs 0.70, P<0.01 ], as well as protein expression level. Positive signal of HMGB1 proteins was not only expressed in nuclear but also in cytoplasm after stimulation. ② Compared with normal group, HMGBI increased the expression of P-STAT1 mRNA and protein at 6, 12, 24 h respectively [0.30, 0.69, 1.05 vs 0.24, P<0.01 ] and [1.34±0.09，1.55±0.16，1.74±0.13 vs 1.00±0.15，P<0.01]. The expression of SOCSI protein increased significantly in HMGB1 group at 6 and 12 hours ( 1.43±0.10 vs 1.58±0.05), but it decreased at 24 hours (1.24±0.15). ③The expression of p-STATI protein was negatively correlated with that of SOCS1 protein. Conclusion HMGB1 appears to be an important mediator in the proliferation of RSC-364 cells, partly by up-regulating the expression and aetivity of p-STAT1.