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ObjectiveTo explore the role of interleukin-6 （IL-6）/signal transducer and activator of transcription 3 （STAT3） pathway in the balance of T helper 17 （Th17）/regulatory T （Treg） cells in ulcerative colitis （UC） with internal dampness-heat accumulation syndrome and the intervention mechanism of Shaoyaotang. MethodA total of 60 SD rats were randomized into blank group （equivalent volume of normal saline）， model group （equivalent volume of normal saline）， western medicine control group （0.42 g·kg-1 mesalazine）， and low-dose （11.1 g·kg-1）， medium-dose （22.2 g·kg-1）， and high-dose （44.4 g·kg-1） Shaoyaotang groups. UC with internal dampness-heat accumulation syndrome was induced in rats with the compound method except for the blank group. The administration lasted 14 days for each group. At 24 h after the last administration， rats were killed and the spleen and colon tissues were separated. The histopathological changes of colon were observed based on hematoxylin and eosin （HE） staining and the levels of interleukin-17 （IL-17） and transforming growth factor-β1 （TGF-β1） in colon tissue were detected by immunohistochemistry （IHC）. Flow cytometry was employed to determine the levels of Th17/Treg cells in the spleen， and Western blot to measure the levels of IL-6 and STAT3 proteins in colon tissue. ResultCompared with the blank group， the model group had lesions such as congestion and erosion， low percentage of spleen Treg cells （P<0.01）， high percentage of Th17 cells （P<0.01）， and high levels of IL-6 and STAT3 proteins in colon tissue （P<0.01）. Compared with the model group， the administration groups showed alleviation of colon injury， high percentage of spleen Treg cells （P<0.05， P<0.01）， low percentage of Th17 cells （P<0.01）， and low levels of IL-6 and STAT3 proteins in colon tissue （P<0.01）. ConclusionShaoyaotang regulates the balance of Th17/Treg by inhibiting the IL-6/STAT3 pathway， thereby relieving the pathological damage of UC rats with internal dampness-heat accumulation syndrome and affecting their immune function.
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Objective@#To explore the effect of high mobility group box-1 protein (HMGB1) on the balance of Th17/Treg in patients with immune thrombocytopenia (ITP).@*Methods@#A total of 30 patients who were first diagnosed as ITP in the Fifth People's Hospital of Datong from July 2017 to April 2018 were selected as the case group, and another 30 healthy volunteers in the corresponding period were taken as the control group. The proportion of Th17 and Treg cells was detected by using flow cytometry, and the concentration of HMGB1, interleukin (IL)-17 and transforming growth factor β (TGF-β) in plasma was tested by using enzyme-linked immunosorbent assay (ELISA). Isolated peripheral blood mononuclear cells (PBMC) were cultured in vitro. After the treatment with recombinant human HMGB1 (rhHMGB1), real-time polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression changes in Treg cell transcription factor intracellular forkhead helix transcription factor 3 (Foxp3) and Th17 cell transcription factor retinoid related orphan receptor γt (RORγt). The differences of indicators in Treg cell transcription factor peripheral blood between the case group and the control group were compared, and the balance correlation between HMGB1 and Th17/Treg was analyzed.@*Results@#Compared with the healthy control group, the proportion of Th17 cells and the expression level of HMGB1 and IL-17 in peripheral blood of ITP patients were increased (all P < 0.01), while the proportion of Treg cells and the level of TGF-β were decreased (all P < 0.01). The proportion of Th17 cells and the expression level of IL-17 and HMGB1 in peripheral blood of ITP patients were positively correlated with the concentration of HMGB1 (all P < 0.01); the proportion of Treg cells and the level of TGF-β were negatively correlated with the expression level of HMGB1 (all P < 0.01). In vitro experiments, the expression of intracellular RORγt mRNA was increased compared with the negative control group (1.50±0.24 vs. 0.93±0.22, t = 9.612, P < 0.01), and the expression of Foxp3 mRNA was decreased compared with the negative control group after the stimulation of PBMC by rhHMGB1 (0.72±0.19 vs. 1.08±0.18, t = 7.387, P < 0.01).@*Conclusion@#The high level of HMGB1 in the peripheral blood of ITP patients induces Th17/Treg imbalance and aggravates inflammatory reactions, which may be an important cause of ITP.
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T helper 17 (Th17) and regulatory T cells (Tregs) are two distinct subsets of T cells that play a key role in the development of autoimmunity and inflammation. Th17 cells are thought to be the key effector T cells that induce inflammatory responses while Tregs inhibit the development of inflammation by regulating effector T cell activity to maintain peripheral immune tolerance. Th17/Treg imbalance can lead to the development of autoimmune diseases. MicroRNAs (miRNAs) are small endogenous non-coding RNA molecules in eukaryotes that play important regulatory roles in the maintenance of homeostasis in the immune system and the development of autoimmune diseases. Abnormal expression of miRNA will result in a broken or dysfunctional balance of differentiation between Th cell subsets, leading to inflammation or autoimmune diseases. This article provides an overview of the research advances in miRNA regulation of Th17/Treg balance to explore the role and clinical significance of miRNAs in Th17/Treg balance and maintenance of immune system balance.
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T helper 17 (Th17) and regulatory T cells (Tregs) are two distinct subsets of T cells that play a key role in the development of autoimmunity and inflammation. Th17 cells are thought to be the key effector T cells that induce inflammatory responses while Tregs inhibit the development of inflammation by regulating effector T cell activity to maintain peripheral immune tolerance. Th17/Treg imbalance can lead to the development of autoimmune diseases. MicroRNAs (miRNAs) are small endogenous non-coding RNA molecules in eukaryotes that play important regulatory roles in the maintenance of homeostasis in the immune system and the development of autoimmune diseases. Abnormal expression of miRNA will result in a broken or dysfunctional balance of differentiation between Th cell subsets, leading to inflammation or autoimmune diseases. This article provides an overview of the research advances in miRNA regulation of Th17/Treg balance to explore the role and clinical significance of miRNAs in Th17/Treg balance and maintenance of immune system balance.
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Objective@#To investigate the effects of artesunate treatment on chronic graft-versus-host disease (cGVHD).@*Methods@#Recipient BALB/c mice received 8 × 106 bone marrow cells with 8×106 spleen cells from B10D2 mice. Artesunate solubilized in acetone was injected intraperitoneally every day at the dose of 1 mg/kg at Day 28 after BMT. The clinical scores, survival and histopathological damage were analyzed. The frequency of Th17 and Tregs in PB and spleens from the mice were evaluated by flow cytometry. In addition, CD4+ T cells from the spleens of mice were cultured in vitro, then stimulated with artesunate, the frequency of Th17 and Tregs in these splenocytes were evaluated by flow cytometry.@*Results@#Artesunate administration diminished clinical and histopathological damage, and improved the survival of cGVHD mice[(46.57±7.83)% vs (55.71±6.99)%, χ2=5.457, P=0.020]; Artesunate contributed to Tregs development [(4.45±0.04)% vs (8.40±0.23)%, t=15.679, P<0.001; (6.62±0.24)% vs (10.48±0.48)%, t=6.587, P=0.003] while decreased Th17 cells [(1.51±0.18)% vs (0.58±0.19)%, t=7.233, P<0.001; (1.48±0.38)% vs (0.71±0.18)%, t=3.653, P=0.011] expressions in both PB and spleens, and decreased the Th17/Treg ratio (0.34±0.05 vs 0.09±0.03, t=7.621, P=0.002; 0.19±0.03 vs 0.06±0.02, t=6.993, P=0.002). Moreover, artesunate suppressed the Th17 cells expressions [(0.82±0.37) % vs (3.39±1.22) %, t=4.044, P=0.007] and contributed to Tregs development [(34.63±1.29) % vs (14.28±1.69) %, t=19.119, P<0.001], and also decreased the Th17/Treg ratio (0.24±0.09 vs 0.02±0.01, t=4.780, P=0.003) in vitro.@*Conclusions@#Artesunate suppressed the Th17 cells expressions and contributed to Tregs development, which provided new sights into the development of a novel drug for cGVHD, e.g., artemisinin.
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Objective To analyze the relationships of high mobility group box 1 protein (HMGB1) with regulatory T cells (Treg), T helper 17 cells (Th17) and cytokine secrtion in peripheral blood of gravidas with preeclampsia(PE),and to investigate the mechanism of HMGB1 in regulating Th17/Treg ratio via receptor for advanced glycation end products (RAGE)-IL-6 pathway. Methods Forty gravi-das with mild(20 cases) and severe(20 cases) PE were recruited as experimental groups,20 heathy gravi-das in the third trimester of pregnancy were enrolled as control group. Concentrations of HMGB1,IL-6,IL-17 and TGF-β in peripheral blood of all subjects were determined by enzyme-linked immunosorbent assay (ELISA). Real-time quantitative PCR(RT-PCR) was used to detect the expression of RAGE at mRNA lev-el in peripheral blood mononuclear cells(PBMCs). The percentages of Treg and Th17 cells were determined by flow cytometry. RT-PCR was performed to analyze changes in the expression of RAGE,IL-6,Foxp3 and RORγt at mRNA level after the PBMCs isolated from 20 garvidas with PE were cultured in vitro and stimula-ted with recombinant human HMGB1 (rhHMGB1). Results The levels of HMGB1,IL-6,Th17 and IL-17 in peripheral blood of gravidas with PE were significantly higher than those in the normal pregnancy group. Moreover,HMGB1 level was positively correlated with IL-6 level and ratios of Th17/Treg and IL-17/TGF-β in preeclamptic pregancies. In vitro stimulation of PBMCs with rhHMGB1 significantly enhanced the expres-sion of RAGE,IL-6 and RORγt at mRNA level, but suppressed the expression of Foxp3 at mRNA level. Conclusion Enriched HMGB1 in plasma shifts the Th17/Treg balance towards Th17 dominance via the RAGE-IL-6 pathway, which exacerbates inflammation and participates in the onset of preeclampsia during pregnancy.
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Inflammatory bowel disease(IBD) is characterized by chronic non-specific inflammatory dis ease.The inflammation caused by abnormal immune response in intestinal mucosa exerts an important role in the pathogenesis of IBD.T helper 17 cells(Th17)are involved in the development of chronic inflammation and auto immune disease,whereas regulatory T cells can inhibit the function of autoimmune,and thus the balance of Th17/Treg balance plays an important role in intestinal homeostasis.Various strategies can be done to modulate Th17/Treg balance,such as antigen-presenting cells,microoganisms,some key molecules,natural compounds,proving a promising therapeutic method for IBD.
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OBJECTIVE: To observe the effects of bone marrow mesenchymal stem cells( BMSCs) on the treatment of pulmonary fibrosis in silicosis mice. METHODS: Specific pathogen free healthy male C57BL/6 mice were randomly divided into control group,silicosis group and treatment group with 10 mice in each group. The mice of the control group were given one intra-tracheal injection of 20. 0 μL 0. 90% sodium chloride solution. The silicosis group and treatment group received one 20. 0 μL( mass concentration 250 g/L) of silica dust suspension. After 4 weeks,mice in treatment group were injected with 250. 0 μL of BMSCs suspension( cell density 2 × 10~9/L) by tail vein and silicosis group injected with 250. 0 μL of 0. 90% sodium chloride solution instead,once a week with continuous treatment for 4 weeks. Control group was not given any treatment. Mice were euthanized two weeks after the last treatment. Pathological sections were observed,pulmonary fibrosis score( Ashcroft scores) was marked. Lung coefficient was measured. Lung tissue hydroxyproline( HYP) level and serum transforming growth factor β1( TGF-β1) level were measured. The level of pulmonary fibrosis was scored and the percentages of T helper cell 17( Th17 cell) and regulatory T cell( Treg cell) of spleen and hilar lymph node( HLN) were measured by flow cytometry. RESULTS: The results of lung histopathological examination showed that the pulmonary fibrosis was severe in silicosis group. Massive collagen fiber accumulation and silicotic nodule were found. In treatment group,fibrosis was mild,little collagen fiber accumulation and silicotic nodule were found. The lung coefficient,Aschcroft scores,lung tissue HYP level,serum TGF-β level and the percentage of Th17 cell of spleen and HLN in silicosis group were higher than that of control group( P < 0. 05),while the above indexes of treatment group were lower than that of silicosis group( P < 0. 01). The percentage of Treg cell of spleen and HLN in silicosis group were lower than that of control group( P < 0. 05),while those indexes of treatment group were higher than that of silicosis group( P < 0. 01).CONCLUSION: BMSCs could effectively alleviate the pulmonary fibrosis in silicosis mice and correct the imbalance of Th17/Treg.