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ObjectiveTo observe the intervention effect of Dahuang Xiezhuo prescription （DHXZ） on inflammation and suppressor of cytokine signaling 3 （SOCS3）/Toll-like receptor 4 （TLR4） pathway in rats with chronic renal failure （CRF）， and to explore its molecular mechanism in alleviating renal inflammatory response. MethodThe 90 male SD rats， 15 were randomly selected as sham group， and the remaining 75 were used as modeling group to replicate CRF rat model by 5/6 nephrectomy. After successful modeling， the rats were randomly divided into model group， DHXZ low-， medium-， high-dose groups （6.825， 13.65， 27.3 g·kg-1） and Niaoduqing Granules group （2.6 g·kg-1）. The drug intervention groups received corresponding drugs by gavage for 8 consecutive weeks. After administration， hematoxylin-eosin （HE） staining and Masson staining were used to observe the morphological changes of rat renal tissue， and blood creatinine （SCr）， blood urea nitrogen （BUN） and blood uric acid （UA） were tested. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum contents of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） and C-reactive protein （CRP）. The mRNA expressions of SOCS3 and TLR4 in renal tissue were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the protein expressions of SOCS3， TLR4， nuclear transcription factor （NF-κB） and myeloid differentiation factor （MyD88） were detected by Western blot. Immunohistochemistry was used to determine the protein expressions of NF-κB， MyD88， NOD-like receptor protein 3 （NLRP3） and melanoma deficiency factor 2 （AIM2）. ResultCompared with the sham group， the model group had a significant inflammatory response in renal tissue， and an increase in blood SCr， BUN， UTP， IL-6， TNF-α and CRP （P<0.05）. The protein and mRNA expressions of SOCS3 in renal tissue of rats in the model group were lower while the protein expressions of TLR4， NF-κB， MyD88， NLRP3 and AIM2 and the mRNA expression of TLR4 were higher than those in the sham group （P<0.05）. Compared with the model group， DHXZ and Niaoduqing granules groups presented markedly reduced inflammatory response in renal tissue and decreased blood SCr， BUN， UTP， IL-6， TNF-α and CRP （P<0.05）. Additionally， DHXZ and Niaoduqing granules up-regulated the protein and mRNA expressions of SOCS3 in renal tissue while down-regulated the protein expressions of TLR4， NF-κB， MyD88， NLRP3 and AIM2 and the mRNA expression of TLR4 （P<0.05）. ConclusionDHXZ can reduce the release and expression of inflammatory factors， inhibit the inflammatory response and improve renal function， and the mechanism may be related to the regulation of SOCS3/TLR4 signaling pathway.
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ObjectiveTo observe the effect of Hedysari Radix polysaccharide （HRP） on the Janus kinase 2 （JAK2）/signal transducer and activator of transcription protein 3 （STAT3） signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group， irbesartan group （irbesartan suspension， 22.75 mg·kg-1）， and high-， medium-， and low-dose HRP groups （HRP suspension， 200， 100， 50 mg·kg-1） according to the body weight， with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage， while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid （UA）， triglycerides （TG）， and total cholesterol （TC） were detected. Periodic acid-Schiff （PAS） staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） were used to detect the protein and mRNA expression levels of JAK2， STAT3， suppressor of cytokine signaling 3 （SOCS3）， and tumor necrosis factor-α （TNF-α） in the kidney. ResultAfter 12 weeks of treatment， compared with the normal group， the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA， TG， and TC levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA， TG， and TC levels （P<0.05， P<0.01）. Compared with the normal group， the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.01）. Compared with the model group， the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2， STAT3， and TNF-α protein and mRNA expression levels （P<0.05， P<0.01）. ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent， and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.
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【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.
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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.
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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.
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OBJECTIVE@#Rheumatoid arthritis (RA) progression is associated with the balance of T-regulatory (Treg) and T-helper 17 (Th17) cells, while the role of microRNAs (miRs) in regulating Treg/Th17 cell balance has not been clarified. This study aimed to assess whether moxibustion could regulate Treg/Th17 cell balance by modulating the miR-221/suppressor of cytokine signaling 3 (SOCS3) axis in the RA mouse model.@*METHODS@#A mouse model of collagen-induced arthritis (CIA) was established in male DBA/1J mice. Twenty-two days after CIA induction, the mice received daily treatment with moxibustion for 12 times. Pathological scores were assessed according to the levels of synovial hyperplasia. The expression levels of cytokines interleukin (IL)-1β, IL-6, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-17 and IL-10 were analyzed in serum by enzyme-linked immunosorbent assay. The cluster of differentiation 4 (CD4+) splenocytes was analyzed by fluorescence-activated cell sorting. The expression levels of RA-related miRs and target genes were subsequently detected, and the target of miR-221 was confirmed by the dual-luciferase reporter assay.@*RESULTS@#It was revealed that moxibustion treatment decreased the pathological scores and downregulated the expression levels of IL-1β, IL-6, TNF-α, IFN-γ and IL-17, while upregulated the expression level of IL-10. The Treg/Th17 cell balance was regulated by moxibustion treatment. The expression level of miR-221 was suppressed by moxibustion treatment. Furthermore, SOCS3 was found as the direct target of miR-221, which mediated the function of moxibustion by regulating the Treg/Th17 cell balance.@*CONCLUSION@#Moxibustion therapy regulated the Treg/Th17 cell balance by modulating the miR-221/SOCS3 axis in the RA mouse model.
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Animals , Male , Mice , Arthritis, Experimental/therapy , Arthritis, Rheumatoid/therapy , Cytokines , Disease Models, Animal , Interleukin-10 , Interleukin-17 , Interleukin-6 , Mice, Inbred DBA , MicroRNAs/genetics , Moxibustion , T-Lymphocytes, Regulatory , Th17 Cells/pathology , Tumor Necrosis Factor-alphaABSTRACT
@#BACKGROUND: Exosomes and exosomal microRNAs have been implicated in tumor occurrence and metastasis. Our previous study showed that microRNA-761 (miR-761) is overexpressed in hepatocellular carcinoma (HCC) tissues and that its inhibition affects mitochondrial function and inhibits HCC metastasis. The mechanism by which exosomal miR-761 modulates the tumor microenvironment has not been elucidated. METHODS: Exosomal miR-761 was detected in six cell lines. Cell counting kit-8 (CCK-8) and transwell migration assays were performed to determine the function of exosomal miR-761 in HCC cells. The luciferase reporter assay was used to analyze miR-761 target genes in normal fibroblasts (NFs). The inhibitors AZD1480 and C188-9 were employed to determine the role of the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in the transformation of cancer-associated fibroblasts (CAFs). RESULTS: In this study, we characterized the mechanism by which miR-761 reprogrammed the tumor microenvironment. We found that HCC-derived exosomal miR-761 was taken up by NFs. Moreover, HCC exosomes affected the tumor microenvironment by activating NFs via suppressor of cytokine signaling 2 (SOCS2) and the JAK2/STAT3 signaling pathway. CONCLUSIONS: These results demonstrated that exosomal miR-761 modulated the tumor microenvironment via SOCS2/JAK2/STAT3 pathway-dependent activation of CAFs. Our findings may inspire new strategies for HCC prevention and therapy.
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ObjectiveTo explore the role of aryl hydrocarbon receptor/suppressor of cytokine signaling 2/nuclear transcription factor-κB (AHR/SOCS2/NF-κB) signaling in the reduction of spinal cord injury (SCI) inflammation with every-other-day fasting (EODF). MethodsA total of 36 healthy male Sprague-Dawley rats were randomly divided into sham group (n = 12), model group (n = 12) and EODF group (n = 12). The latter two groups were established C5 spinal cord hemi-clamp model. The EODF group received EODF, namely fasting and feeding per 24 hours (water free), for two weeks after operation. They were assessed with Grooming Test before operation, and one, seven and 14 days after operationt, respectively. Pathological injury of spinal cord was observed with HE staining 14 days after operation, while the expression of AHR, SOCS2 and NF-κB proteins were detected with Western blotting, and the mRNA of AHR, SOCS2, inhibitor α of NF-κB (IκBα), NF-κB and tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) were measured with reverse transcription real-time quantitative polymerase chain reaction. ResultsCompared with the sham group, the Grooming Test score decreased in the other two groups (P < 0.05), and cavity, edema and structural disorder appeared in the spinal cord. Compared with the model group, the pathology of EODF group improved, and the Grooming Test score increased (P < 0.05). The mRNA and protein expression of AHR and SOCS2 increased (P < 0.05), the mRNA and protein expression of NF-κB decreased (P < 0.05), the mRNA expression of IκBα increased (P < 0.05), and the mRNA expression of TNF-α and IL-6 decreased (P < 0.05). ConclusionEODF may promote the recovery of motor function of forelimb in rats with SCI, and relieve damage and inflammation, which may associate with the activation of AHR/SOCS2/NF-κB signaling pathway.
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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.
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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 (
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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.
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OBJECTIVE: To investigate the effect of acupuncture plus moxibustion on learning-memory ability and expression of hippocampal Janus kinase-2 (JAK2)/signal transducer and activator of transcription-3 (STAT3)/suppressors of cytokine signaling-3 (SOCS3) signaling in Alzheimer's Disease (AD) rats, so as to reveal their mechanisms underlying improvement of AD. METHODS: A total of 60 SD rats were randomly divided into four groups：normal control, sham-operation, model and acupuncture-moxibustion (Acu-moxi, n＝15 in each group) groups. The AD model was established by microinjection of β-amyloid 1-42(Aβ1-42，5 µL)into the bilateral hippocampus. Seven days after modeling, Acu-moxi intervention was given. After insertion of acupuncture needles into "Baihui" (GV20) and bilateral "Shenshu" (BL23) and manipulating them for a while, the needles were then retained for 15 min, when, the mild moxibustion was performed at the same time. The treatment was conducted once daily, 5 times a week for consecutive 4 weeks. After the treatment, Morris water maze test was used to detect the animals' learning-memory ability. Immunohistochemistry and Western blot were respectively used to detect the number of positive cells and protein expression levels of JAK2, STAT3 and SOCS3 in the hippocampus tissue. RESULTS: Following modeling and compared with the normal control and sham-operation groups, the average escape latency was significantly prolonged (P<0.01), and the number of the original platform crossing and the residence time in the platform quadrant were significantly shortened in the model group (P<0.01). The numbers of hippocampal JAK2- and STAT3-positive cells and expression levels of hippocampal JAK2 and STAT3 proteins were significantly increased (P<0.01), and the number of hippocampal SOCS3-positive cells as well as the expression of SOCS3 protein significantly decreased in the model group relevant to the normal control and sham-operation groups (P<0.01). After the intervention, the average escape latency was significantly shortened (P< 0.01), and the number of the original platform crossing and the residence time in the platform quadrant were significantly increased in the Acu-moxi group (P<0.01), and the expression levels of JAK2 and STAT3 were significantly down-regulated and that of SOCS3 was considerably up-regulated in the Acu-moxi group relevant to the model group (P<0.01)．. CONCLUSION: Acu-moxi intervention can improve the learning-memory ability in AD rats, which is associated with its functions in inhibiting hippocampal JAK2/STAT3 signaling and up-regulating SOCS3 (a negative feedback factor) protein level.
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Objective: To illuminate the changes in expression of suppressor of cytokine signaling (SOCS) and the correlation between SOCS expression and cholesterol levels in atherosclerosis (AS). Methods: A mouse AS model was established in apolipoprotein-deficient (ApoE-/-) mice. The mice were fed either with a chow or high-fat diet. The mRNA and protein expressions of SOCS1 and SOCS3 in plaque and vessels were determined at different time points. Furthermore, SOCS1 and SOCS3 mRNA expressions were detected in the peripheral blood mononuclear cells (PBMCs) obtained from 18 male subjects with no coronary heart disease (non-CHD). Results: The expression of SOCS1 in the ApoE-/- mice increased first and then decreased and the high-fat diet accelerated the appearance of the peak. The expression of SOCS3 increased with the increase of feeding duration, and this trend was more pronounced in the mice fed with high-fat diet. SOCS1/CD68 and SOCS3/CD68 showed the opposite trend in expression with the increased duration of high-fat diet. Interleukin-6 (IL-6) expression in the aorta of the ApoE-/- mice fed with high-fat diet also increased with the increased feeding duration. In the non-CHD group, the total serum cholesterol level was positively correlated with SOCS3 mRNA expression in the PBMCs (r=0.433, P=0.012). Conclusion: These results demonstrate the differential expressions of SOCS1 and SOCS3 in AS and suggest that SOCS3, together with IL-6, may promote the formation and development of AS.
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Objective@#To compare the effect of insulin-like growth factor-1 receptor (IGF-1R) inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease (DKD) mice.@*Methods@#Twenty-four male C57BL/6 mice were selected. After 1 week of adaptive feeding, 6 rats were randomly selected as the control group. The other mice were intraperitoneally injected with streptozotocin (30 mg/kg) after 8 weeks of high-fat and high-sugar feeding. After 72 h, the type 2 diabetes mellitus (DM) models were successfully established if random blood glucose was greater than 16.7 mmol/L. After 8 weeks, if the proteinuria of DM mice increased, the DKD models were successful. DKD mice were divided into 3 groups by random number remainder method: DKD group (n=6), DKD+insulin group (insulin group, n=6, subcutaneous injection of 1-2 U/d insulin) and DKD+IGF-1R inhibitor (IGF-1R inhibitor group, n=6, administered with 30 mg·kg-1·d-1 IGF-1R inhibitor). They were continuously treated for 8 weeks. Random blood glucose was tested by glucometer. Blood and urine were collected, and biochemical indicators, such as serum creatinine, urea nitrogen and urine protein were measured by biochemical analyzer. Renal pathological changes were detected by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS). Suppressor of cytokine signaling 2 (SOCS2) mRNA and insulin-like growth factor-1 (IGF-1) mRNA were detected by in situ hybridization. The protein expressions of SOCS2, F4/80, Toll-like receptor 4 (TLR4) and CD68 were detected by immunohistochemistry.@*Results@#Compared with the control group, blood glucose, serum creatinine, serum urea nitrogen and urinary protein excretion rate were significantly higher in DKD mice (all P<0.05), and CD68+ cells number, F4/80+ cells number and the expression of TLR4 in the tubulointerstitial of DKD mice were significantly higher (all P<0.05). After intervention with insulin or IGF-1R inhibitor, serum creatinine, serum urea nitrogen and urinary protein excretion rate of DKD mice were significantly reduced (all P<0.05). Insulin intervention could significantly reduce blood glucose in mice (P<0.05), but had no significant effect on macrophages. Although IGF-1R inhibitor did not significantly reduce blood glucose, it could significantly reduce the number of CD68, F4/80 positive cells and the expression of TLR4 protein in renal interstitium of DKD mice (all P<0.05). Compared with the DKD group, insulin intervention significantly reduced the expression of IGF-1 protein and mRNA (both P<0.01), and increased the expression of SOCS2 mRNA and protein (both P<0.01). And the expression of SOCS2 protein was correlated with the number of F4/80+ cells in insulin group (R2=0.8461, P=0.005). However, IGF-1R inhibitors had no significant effect on SOCS2 expression, but had better inhibition of macrophage infiltration.@*Conclusion@#IGF-1R inhibitor has a better inhibitory effect on DKD renal interstitial macrophage infiltration than insulin. The mechanism may be related to the fact that IGF-1R inhibitor does not up-regulate SOCS2 expression, whereas insulin up-regulates SOCS2 expression to activate some potential pathways.
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Objective To compare the effect of insulin-like growth factor-1 receptor (IGF-1R) inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease (DKD) mice. Methods Twenty-four male C57BL/6 mice were selected. After 1 week of adaptive feeding, 6 rats were randomly selected as the control group. The other mice were intraperitoneally injected with streptozotocin (30 mg/kg) after 8 weeks of high-fat and high-sugar feeding. After 72 h, the type 2 diabetes mellitus (DM) models were successfully established if random blood glucose was greater than 16.7 mmol/L. After 8 weeks, if the proteinuria of DM mice increased, the DKD models were successful. DKD mice were divided into 3 groups by random number remainder method: DKD group (n=6), DKD+insulin group (insulin group, n=6, subcutaneous injection of 1-2 U/d insulin) and DKD+IGF-1R inhibitor (IGF-1R inhibitor group, n=6, administered with 30 mg·kg-1·d-1 IGF-1R inhibitor). They were continuously treated for 8 weeks. Random blood glucose was tested by glucometer. Blood and urine were collected, and biochemical indicators, such as serum creatinine, urea nitrogen and urine protein were measured by biochemical analyzer. Renal pathological changes were detected by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS). Suppressor of cytokine signaling 2 (SOCS2) mRNA and insulin-like growth factor-1 (IGF-1) mRNA were detected by in situ hybridization. The protein expressions of SOCS2, F4/80, Toll-like receptor 4 (TLR4) and CD68 were detected by immunohistochemistry. Results Compared with the control group, blood glucose, serum creatinine, serum urea nitrogen and urinary protein excretion rate were significantly higher in DKD mice (all P<0.05), and CD68+ cells number, F4/80+ cells number and the expression of TLR4 in the tubulointerstitial of DKD mice were significantly higher (all P<0.05). After intervention with insulin or IGF-1R inhibitor, serum creatinine, serum urea nitrogen and urinary protein excretion rate of DKD mice were significantly reduced (all P<0.05). Insulin intervention could significantly reduce blood glucose in mice (P<0.05), but had no significant effect on macrophages. Although IGF-1R inhibitor did not significantly reduce blood glucose, it could significantly reduce the number of CD68, F4/80 positive cells and the expression of TLR4 protein in renal interstitium of DKD mice (all P<0.05). Compared with the DKD group, insulin intervention significantly reduced the expression of IGF-1 protein and mRNA (both P<0.01), and increased the expression of SOCS2 mRNA and protein (both P<0.01). And the expression of SOCS2 protein was correlated with the number of F4/80 + cells in insulin group (R2=0.8461, P=0.005). However, IGF-1R inhibitors had no significant effect on SOCS2 expression, but had better inhibition of macrophage infiltration. Conclusion IGF-1R inhibitor has a better inhibitory effect on DKD renal interstitial macrophage infiltration than insulin. The mechanism may be related to the fact that IGF-1R inhibitor does not up-regulate SOCS2 expression, whereas insulin up-regulates SOCS2 expression to activate some potential pathways.
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Objective Dendritic cells (DCs), helper T cells 17 (Th17) and regulatory T cells (Treg) are closely related to the pathogenesis of chronic obstructive pulmonary disease (COPD). This study aimed to investigate the changes of Th17- and Treg-related cytokines in the bronchoalveolar lavage fluid (BALF) of COPD mice after DC-based adoptive immunotherapy with over-expressed suppressor of cytokine signaling protein 1 (SOCS1) and provide some new ideas for the treatment of COPD. Methods A total of 48 male C57BL/6 mice were randomly divided into 5 groups: healthy control, COPD model control, immature DC (imDC), DC-SOCS1 1×106, and DC-SOCS1 2×106. The healthy controls were exposed to air and fed normally, the COPD model controls injected with normal saline at 0.5 mL/ on the first day of modeling by fumigation, the mice of the imDC group injected via the tail vein with 1 ×106 imDCs, and those of the DC-SOCS1 groups injected with 1 ×106 or 2 ×106 DCs with over expressed SOCS1, all via the tail vein on the 1st and 7th day of modeling. Then the lung tissues were collected from the mice for preparation of paraffin sections and HE staining, and ELISA was employed for determination of the levels of Th17-related IL-17 and IL-23 and Treg-related IL-10 and TGF-β in the BALF of the model mice. Results Compared with the COPD model controls, the mice in the imDC, DC-SOCS1 1×106 and DC-SOCS1 2×106 groups showed significantly decreased levels of IL-17 on the 1st day ([78.87 ± 1.08] vs [46.46 ± 0.77], [34.09 ± 3.98] and [24.12 ± 0.57] pg/mL, P < 0.05) and 7th day after modeling ([78.87 ± 1.08] vs [55.69 ±0.35], [35.65 ± 0.54] and [27.00 ± 0.58] pg/mL, P < 0.05), and IL-23 on the 1st day ([200.62 ± 0.65] vs [150.19 ± 0.53], [121.09 ± 0. 53] and [70.21 ± 0.91] pg/mL, P < 0.05) and 7th day ([200.62 ± 0.65] vs [167.70 ± 1.73], [136.34 ± 0.90] and [99.35 ± 1.83] pg/mL, P < 0.05), but remarkably increased levels of IL-10 on the 1st day ([39.46 ± 3.88] vs [50.74 ± 1.77], [58.71 ± 3.84] and [70.12 ± 2.62] pg/mL, P < 0.05) and 7th day ([39.46 ± 3.88] vs [44.56 ± 2.63], [54.78 ± 1.43] and [63.00 ± 2.57] pg/mL, P < 0.05), TGF-β on the 1st day ([24.98 ± 0.43] vs [36.46 ± 0.98], [42.40 ± 0.62] and [50.55 ± 0.53] pg/mL, P < 0.05) and 7th day ([24.98 ± 0.43] vs [33.27 ± 0.92], [40.12 ± 0.83] and [44.98 ± 0.52] pg/mL, P < 0.05). The contents of IL-17 and IL-23 were markedly lower while those of IL-10 and TGF-β higher in the DC-SOCS1 1×106 than in the imDC group (P < 0.05), and the levels of the former two significantly higher and those of the latter two lower in the DC-SOCS1 2×106 than in the DC-SOCS1 1×106 group (P < 0.05). Conclusion Transfusion of DCs with over-expressed SOCS1 can inhibit the secretion of Th17-related cytokines in COPD, and the effect is better than that of imDCs alone and related to the concentration and time.
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Objective To investigate the relationship among serum heat shock protein 70 (HSP70), suppressor of Cytokine Signaling-3 ( SOCS-3) and immune factor in pregnant women with hypertension,and to analyze the diagnostic value of the two indicators. Methods Eighty-six pregnant women with hypertension who were treated in the Second People′s Hospital of Yichang from January 2016 to February 2018 were selected,according to the severity of the disease,they were divided into pre-eclampsia (51 cases) and severe pre-eclampsia group (35 cases),another 40 normal pregnant women in the same period were selected as control group. The serum levels of HSP70 and SOCS-3,plasma immunoglobulin A (IgA), immunoglobulin M ( IgM), immunoglobulin G ( IgG), complement C3 and complement C4 levels were detected in each group,the correlation between serum HSP70 and SOCS-3 levels and immune factors were analyzed. ROC curve was used to analyze the predictive value of serum HSP70 and SOCS-3 in hypertensive disorder complicating pregnancy. Results The serum HSP70 of pre eclampsia and severe preeclampsia group was ( 3. 92 ± 0. 35 ) μg/L, the serum HSP70 of eclampsia group was ( 6. 45 ±0. 78) μg/L,which were significantly higher than that of the control group ( 0. 36 ± 0. 07) μg/L, the differences were statistically significant (t＝63. 272,49. 202,P＜0. 05),the serum SOCS-3 of pre eclampsia and severe preeclampsia group was (0. 22±0. 08) ng/L,the serum SOCS-3 of eclampsia group was (0. 10 ±0. 03) ng/L,which were significantly lower than ( 0. 62 ± 0. 11) ng/L that of the control group ( 0. 62 ±0. 11) ng/L,the differences were statistically significant (t＝－20. 078,－27. 079,P＜0. 05),the serum HSP70 of eclampsia group was significantly higher than that of the pre eclampsia and severe preeclampsia group,the difference was statistically significant (t＝－20. 402,P＜0. 05),the serum SOCS-3 of eclampsia group was significantly lower than that of the pre eclampsia and severe preeclampsia group,the difference was statistically significant ( t＝8. 462, P＜0. 05) . The plasma IgM was ( 1. 83 ± 0. 56) g/L, IgG was ( 7. 94 ±1. 34) g/L,complement C3 was (0. 95±0. 08) g/L,complement C4 was (0. 24±0. 08) g/L in the pre eclampsia and severe preeclampsia group, the plasma IgM was ( 1. 42 ± 0. 58 ) g/L, IgG was ( 5. 23 ±1.13) g/L,complement C3 was (0.73±0.12) g/L,complement C4 was (0.13±0.05) g/L in the eclampsia group,the plasma IgM was (2. 55±0. 53) g/L,IgG was (11. 04±2. 15) g/L,complement C3 was (1. 28 ±0. 15) g/L,complement C4 was (0. 35±0. 08) g/L in the control group (IgM:t＝－6. 232,－8. 815, P＜0. 05;IgG: t＝－8. 426,－14. 340, P＜0. 05; C3: t＝－13. 470,－17. 364, P＜0. 05; C4: t＝－6. 510,－14. 040,P＜0. 05),the plasma levels of IgM,IgG,complement C3 and complement C4 in eclampsia group were significantly lower than those in pre eclampsia and severe preeclampsia group ( t＝3. 288,－9. 805, 10. 209,7. 217,P＜0. 05). Pearson correlation analysis showed,there was a negative correlation among serum HSP70 and plasma IgM,IgG,complement C3 and complement C4 in hypertensive women with gestational hypertension (r＝－0. 446,－0. 537,－0. 426,－0. 428,P＜0. 05),serum SOCS-3 was positively correlated with plasma IgM,IgG,complement C3 and complement C4 (r＝0. 423,0. 507,0. 416,0. 407,P＜0. 05),there was no correlation among serum HSP70, SOCS-3 and plasma IgA in pregnant women with hypertension ( r＝－0. 082,0. 093,P＞0. 05). The area under the ROC curve of HSP70 was 0. 821,and the critical value of the prediction was more than 0. 89 μg/L,the sensitivity of HSP70 to pregnant women with hypertension was 86. 3%,the specificity was 76. 4%,the area under the ROC curve of SOCS-3 was 0. 759,the critical value of the prediction was less than 0. 035 ng/L,and the sensitivity of SOCS-3 to pregnant women with hypertension was 79. 4%,and the specificity was 71. 6%. Conclusion The abnormal increase of serum HSP70 and abnormal decrease of SOCS-3 in pregnant women with hypertension, maternal serum HSP70 and SOCS-3 levels are closely related to immune factors,the serum HSP70 and SOCS-3 may be used as early predictors of hypertensive disorder complicating pregnancy.
ABSTRACT
Triptolide( TP) is isolated from the traditional Chinese medicine Tripterygium wilfordii,which exhibits notable immuneregulative effect. Th17 cells involve in inflammatory response and Treg cells contribute to immune tolerance. They both play an important role in immune response. Previous studies have investigated that TP induced hepatic Th17/Treg imbalance. However,the effect of TP on spleen Th17/Treg cells remains unclear. Therefore,the aim of present study was to investigate the effect of TP on Th17/Treg cells in spleen. In this study,the effect of TP on the proliferation of splenic lymphocyte was detected by cytotoxicity test in vitro. After different concentrations of TP( 2. 5,5,20,40 nmol·L~(-1)) were given to splenic lymphocyte,cytokines secreted from the supernatant of splenic lymphocyte were detected by cytometric bead array,and the expression of suppressor of cytokine signaling( SOCS) mRNA was detected by qRT-PCR. Female C57 BL/6 mice were continuously observed for 24 h after treatment of 500 μg·kg-1 TP. The effects of TP on the splenic tissue structure and the percentage of Th17/Treg cells were examined. The results showed that the IC50 of TP was19. 6 nmol·L~(-1) in spleen lymphocytes. TP inhibited the secretion of IL-2 and IL-10 and induced the expression of SOCS-1/3 mRNA in spleen lymphocytes at the dosage of 2. 5 and 5 nmol·L~(-1) after 24 h in vitro. Administration of TP at dosage of 500 μg·kg-1 had no significant spleen toxicity in vivo. TP treatment increased the percentage of Th17 cells after 12 h and inhibited the proportion of Treg cells after 12 and 24 h. In conclusion,TP reduced the secretion of IL-2 and IL-10 through SOCS-1/3 signaling pathway,thereby induced the percentage of Th17 cells and inhibited the percentage of Treg cells.
Subject(s)
Animals , Female , Mice , Cytokines , Metabolism , Diterpenes , Pharmacology , Epoxy Compounds , Pharmacology , Mice, Inbred C57BL , Phenanthrenes , Pharmacology , Signal Transduction , Spleen , Cell Biology , Suppressor of Cytokine Signaling 1 Protein , Metabolism , Suppressor of Cytokine Signaling 3 Protein , Metabolism , T-Lymphocytes, Regulatory , Cell Biology , Th17 Cells , Cell BiologyABSTRACT
IL-15 is a cytokine of the common γ-chain family that is critical for natural killer (NK), invariant natural killer T (iNKT), and CD8 memory T cell development and homeostasis. The role of IL-15 in regulating effector T cell subsets, however, remains incompletely understood. IL-15 is mostly expressed by stromal cells, myeloid cells, and dendritic cells (DCs). Whether T cells themselves can express IL-15, and if so, whether such T cell-derived IL-15 could play an autocrine role in T cells are interesting questions that were previously addressed but answered with mixed results. Recently, three independent studies described the generation of IL-15 reporter mice which facilitated the identification of IL-15-producing cells and helped to clarify the role of IL-15 both in vitro and in vivo. Here, we review the findings of these studies and place them in context of recent reports that examined T cell-intrinsic IL-15 expression during CD4 effector T cell differentiation.
Subject(s)
Animals , Humans , Mice , Cell Differentiation , Dendritic Cells , Homeostasis , In Vitro Techniques , Inflammation , Interleukin-15 , Memory , Myeloid Cells , Receptors, Cytokine , Stromal Cells , T-Lymphocyte Subsets , T-Lymphocytes , Th17 CellsABSTRACT
Objective To analyze the relationship between the JAK2V617F mutation and the expressions of phosphorylated Janus kinase 2 (p-JAK2), suppressor of cytokine signaling 1 (SOCS1), and SH2-containing protein tyrosine phosphatase 1 (SHP1) in JAK2V617F mutation positive myeloproliferative neoplasm (MPN) tissues, and to investigate the effects of JAK2 inhibitor ruxolitinib on regulating the proliferation of JAK2V617F mutation positive human erythroleukemia cell lines HEL and the expressions of SOCS1 and SHP1 in HEL cells. Methods A total of 48 patients with JAK2V617F mutation positive MPN (MPN group) and 24 patients with anemia (control group) in Hebei General Hospital and The First People’s Hospital of Baoding from Jul. 2012 to Aug. 2016 were enrolled in this study. The protein expressions of p-JAK2, SOCS1 and SHP1 in bone marrow biopsies (BMBs) were detected by immunohistochemistry. The correlations between JAK2V617F mutation level and the protein expressions of SOCS1 and SHP1 were analyzed by Spearman rank correlation analysis. HEL cells were treated with ruxolitinib at different concentrations (50, 100, 250, 500 and 1 000 nmol/L), and the viability of cells was determined by CCK-8 assay. The JAK2V617F mutation levels in MPN tissues and HEL cells and the mRNA expressions of JAK2, SOCS1 and SHP1 in HEL cells were detected by qPCR. The protein expressions of JAK2, SOCS1 and SHP1 in HEL cells were detected by Western blotting analysis. Results The ratio of JAK2V617F/JAK2 was (57.33±20.82)% in the MPN group and was zero in the control group. The protein expressions of p-JAK2, SOCS1 and SHP1 in BMBs of MPN patients were significantly different from those in the control group (all P0.01). The protein expressions of SOCS1 and SHP1 were negatively correlated with the mutation level of JAK2V617F (r＝－0.648, －0.692; P0.05). The expressions of SOCS1 and SHP1 in MPN patients with JAK2V617F/JAK250% were significantly higher than those in MPN patients with JAK2V617F/JAK2≥50% (P0.01), while the expression of p-JAK2 was significantly lower than that in MPN patients with JAK2V617F/JAK2≥50% (P0.01). After treatment with 250 nmol/L ruxolitinib for 24 h, 48 h, and 72 h, the viabilities of HEL cells were (60.06±3.87)%, (52.05±2.88)%, and (36.43±2.01)%, respectively. With the increase of ruxolitinib concentrations, the mRNA and protein expressions of JAK2 and the protein expression of p-JAK2 were gradually decreased (P0.01, P0.05), while the mRNA and protein expressions of SOCS1 and SHP1 were gradually increased (all P0.01). Conclusion Ruxolitinib can inhibit the expressions of JAK and the phosphorylation of JAK in HEL cells, enhance the expressions of SOCS1 and SHP1, and reduce the viability of HEL cells.