Scutellarin alleviates complete freund’s adjuvant-induced rheumatoid arthritis in mice by regulating the Keap1/Nrf2/HO-1 pathway

Scutellarin(SCU)is a herbal flavonoid glucuronide with multiple pharmacological activities,including antioxidant,anti-inflammation,vascular relaxation,anti-platelet,and myocardial protection.However,the effect of SCU on complete Freund’s adjuvant(CFA)-induced rheumatoid arthritis(RA)had not been studied.In this study,we investigated the beneficial effects of SCU in the CFA-induced RA mice model and the anti-arthritic activity was evaluated by paw edema.Enzyme-linked immunosorbent assay(ELISA)was carried out to evaluate the plasma levels of immunoglobulin(Ig)G,IgE,tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,receptor activator of nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG).Histological slides were prepared from the harvested paws of mice to determine the pathological changes in the joints.The proportions of T helper type 1(Th1)and T helper type 2(Th2)cells of CD4+T lymphocyte subsets were analyzed by flow cytometry.The expression of Kelch-like ECHassociated protein 1(Keap1),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)was analyzed using real-time quantitative PCR(RT-qPCR)and western blotting assays.The present study demonstrated that SCU prevented CFA-induced RA,and inhibited the expression of inflammation factors,IgG,IgE,TNF-α,IL-1β,and IL-6.While SCU also reduced the RANKL level,it increased OPG expression in RA mice.The Th1/Th2 ratio was significantly lower in mice treated with SCU.Additionally,HO-1 expression was reduced while the expression of Keap1 and Nrf2 was elevated following SCU treatment.Results provide preliminary evidence to employ SCU in arthritis treatment which might be related to the regulation of Th1/Th2 balance and the Keap1/Nrf2/HO-1 pathway.

Impact of Bisphenol A (BPA) and Free Fatty Acids (FFA) on Th2 Cytokine Secretion from INS-1 Cells

Bisphenol A (BPA) is used in huge amounts for many plastic products and is a hormone (estrogen) disrupting agent. BPA as well as FFAs may be deleterious for the immune system. The aim was to identify Th2 cytokines and some of their signal transduction mechanisms in INS-1 cells, an insulin secreting cell line. Screening using a proteome profile indicated an increase of IL-1, IL-2, IL-4, IL-6, IL-10, IL-13 and IL-17 by BPA. Also FFAs (in combination with LPS) were positive. In detailed quantitative measurements, these results were confirmedly indicating a complex array of pro-and anti-inflammatory potential. The interaction of BPA with 17β-estradiol was non-additive with respect to IL-4 and IL-6 release and additive with respect to FFA interaction indicating same and different mechanisms of action, respecttively. As signal transduction PI3K (Wortmannin-sensitive) and STAT-3/6 (Tofacitinib-sensitive) are involved in various effects, INS-1 cells release several cytokines due to BPA and FFA attack which may be involved in disturbance of glucose homoeostasis and type 1 diabetes.

T Lymphocytes and Th1/Th2 Cytokines in Peripheral Blood of Patients with Recurrent Genital Herpes

Objective: This study analyzed the T lymphocytes and Th1/Th2 type cytokine profile shift in the peripheral blood ofpatients with recurrent genital herpes (RGH). Methods: Immunofluorescent staining or cell surface antigenand intracellular cytokines(IL-2、IL-4、IL-12、IFN-r)inperipheral blood from 20 RGH patients and 10 controls wereanalyzed using flow cytometric techniques. Results: RGH patients had signiflcantly lower levels ofCD3^+T cells, CD4^+T cells and CD4^+ T/ CD8^+ T cells ratiocompared to control levels (P<0.001), and IL-2-producing,IFN-r-producing and IL-12-producing T cells were increasedin RGH patients (CD4^+T: P<0.001, CD8^+T: P<0.05respectively), whereas IL-4-producing T cells were increased inRGH patients compared to controls (CD4^+T: P<0.05; CD8^+T:P<0.001 respectively). Conclusions: RGH patients have T lymphocyte subsetvariations and Th1/Th2 cytokine changes. The increase in Th2cells Th1/Th2 imbalance may have important implications forRGH pathogenesis.

Analysis of Th1/Th2 Response Pattern for Erythrodermic Psoriasis

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis（EP） is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells（PBMCs） derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris（PV） patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls（P〈0.05）. Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients（P〈0.01）. The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

Influence of the invasion of peripheral blood mononuclear cells by hepatitis B virus on immune response of the patients with chronic hepatitis B

Objective:To exploretheinfluenceof HBVinvasionintoperipheralbloodmononuclearcells(PBMC)on theimmuneresponseof patientswithchronichepatitisB.Method s:Thecytokinelevelsintheculturesupernatantof PBMCfrom56patientswithchronichepatitisB weredeterminedby ELISA,andPCRwasemployedto amplifythe HBVDNA.Results:Thelevelsof IFN-γinpatientswithhepatitisB waslowerthanthosetof thecontrol,butthe differencewasnotstatisticallysignificant,whilethelevelsof IL-4weresignificantlyhigherthanthoseof thecontrol(P<0.01).Theserumlevelsof HBVDNAwerenegativelycorrelatedwiththatof IFN-γin culturesupernatantsof PBMC.Thirty-fivepatientspositiveof HBVDNA inthePBMCswereidentifiedfrom56patientswithhepatitisB,andtheirIFN-γlevelprovedto be significantlydifferent.Conclusions:Th2cell-mediatedimmuneresponseis predo-minantin chronichepatitisB whichis associatedwiththechronicityof HBVinfection.HBVinvasionintothe PBMCsmayaffectTh1andTh2cell-mediatedimmuneresponseof thepatientswithchronichepatitisB.

Wang-Bi tablet, a patented Chinese medicine, maintains the balance of Th1/Th2 in mice with collagen-induced arthritis

OBJECTIVE: To investigate the pharmacological mechanism of Wang-Bi tablets(WBTs), a Chinese patented medicine, in rheumatoid arthritis(RA) using mice with collagen-induced arthritis(CIA).METHODS: A mouse model of CIA was induced using bovine type Ⅱ collagen. WBT treatment was administered and efficacy was evaluated. The levels of interferon-γ(IFN-γ), interleukin-2(IL-2), and interleukin-4(IL-4) were examined using an enzyme-linked immunosorbent assay, and the proportions of Th1 and Th2 were detected using flow cytometry. T-bet and GATA-binding protein 3(GATA3)expression were demonstrated using Western blot analysis.RESULTS: Paw swelling and the arthritis index decreased significantly following WBT treatment. Histopathological analysis revealed markedly alleviated damage to synovium tissue in the WBT and methotrexate treatment groups. WBT regulated the production of IFN-γ, IL-2, and IL-4 and modulated Th1 and Th2 cell populations, which might have been induced by the attenuation of Th1 and Th2 cell differentiation through a decrease in the expression of T-bet and an increase in the expression of GATA3 in the synovial tissue in CIA mice.CONCLUSION: These results indicate that WBT may produce a therapeutic effect on CIA through maintaining the balance of Th1/Th2 cells, which could result in a decrease in the autoinflammatory disorder observed in RA.

The relationship between Th1/Th2-type cells and disease activity in patients with systemic lupus erythematosus

Obejctive To investigate the imbalance of Th1/Th2 type cytokines in patients with systemic lupus erythematosus (SLE) and its relation to disease activity Methods Intracellular cytokines were determined by flow cytometry following whole blood culture Results Patients with systemic lupus erythematosus disease activity index (SLEDAI) >10 had statistically significantly fewer CD4 + or CD8 + T cells producing IFN γ than patients with SLEDAI =0, SLEDAI 1-10 or healthy controls ( P <0 01, P <0 01 or P <0 05, respectively) Patients with SLEDAI>10 also had decreased ratio of IFN γ/IL 4 positive CD4 + or CD8 + T cells, compared with patients with SLEDAI =0, SLEDAI 1-10 or healthy controls ( P <0 05) The decreased Th1 or Tc1 cells and the ratios of IFN γ: IL 4 positive CD4 + T cells were significantly correlated with disease activity ( P <0 05) Conclusion SLE is characterized by an imbalance of Th1/Th2 and Tc1/Tc2 cytokines The decreased Th1 or Tc1 cells and the Th1/Th2 ratio are related to disease activity

Antisense-Induced Blockade of GATA-3 Expression Could Inhibit Th2 Excursion of Tumor Cells in vitro and in vivo

Previous studies have shown that tumor cells predominantly express Th2 type cytokines and transcription factors. GATA-3, as a Th2-specific transcription factor, plays a central role in positive-regulating Th2 development. So whether the expression of GATA-3 in tumor cells has any effect on tumor development is a question of interest. In the present study, we inhibited the expression of GATA-3 in tumor cells through antisense RNA blockade technique, and observed its effects on tumor in vitro and in vivo. Our results showed that antisense GATA-3 treatment could inhibit the expression of TNF-α and Th2 cytokines in tumor cells, and antisense-induced blockade of GATA-3 could also depress tumor growth in tumor-bearing mice. We suggest that the ratio of T-bet/GATA-3 can be evaluated as a more important marker of the status of Thl/Th2 type. And our results might provide some evidence about the molecular regulatory mechanisms in tumor cell development. Cellular ＆ Molecular Immunology. 2005 ;2（3 ）： 189-196.

类风湿性关节炎患者外周血Th1/Th2细胞因子与继发骨质疏松症的关系研究

目的探讨类风湿性关节炎患者外周血辅助性T细胞(helper T cell,Th)1/Th2细胞因子与继发骨质疏松症的关系。方法选取2018年9月至2020年8月在北京市石景山医院接受诊治的96例类风湿性关节炎患者作为研究组,根据患者骨密度降低程度将其分为骨质疏松症继发组(60例)和未继发组(36例),另外招募同期84例健康志愿者作为对照组。两组均采用酶联免疫吸附试验法(enzyme linked immunosorbent assay,ELISA)测定外周血Th1/Th2相关细胞因子干扰素-γ(interferon-γ,INF-γ)、白细胞介素(interleukin,IL)-4水平,应用双能X线骨密度检查仪检查骨密度。分别对比分析研究组与对照组、继发组与未继发组的INF-γ、IL-4水平及INF-γ/IL-4,腰椎L1-L4、髋部骨密度表达情况。采用Pearson相关分析研究组外周血Th1/Th2类细胞因子水平与骨密度的相关性;采用Logistic多元回归分析类风湿性关节炎外周血Th1/Th2细胞因子与继发骨质疏松症的关系。结果研究组的外周血INF-γ水平、INF-γ/IL-4均高于对照组,IL-4水平、腰椎骨密度、髋骨骨密度均低于对照组,差异均有统计学意义[(8.55±1.27)μg/L比(5.06±0.94)μg/L,(18.34±3.17)比(4.87±0.85),(0.41±0.07)μg/L比(0.84±0.15)μg/L,(0.96±0.16)g/cm3比(1.15±0.21)g/cm3,(0.82±0.15)g/cm3比(0.89±0.16)g/cm3,t值分别为20.71、37.76、25.14、6.87、3.03,P值均<0.05]。继发组外周血INF-γ水平、INF-γ/IL-4均高于未继发患者,IL-4水平、腰椎骨密度、髋骨骨密度均低于未继发组,差异均有统计学意义[(9.37±1.34)μg/L比(6.14±1.16)μg/L,(21.67±4.12)比(16.33±2.96),(0.37±0.07)μg/L比(0.42±0.08)μg/L,(0.89±0.17)g/cm3比(1.02±0.18)g/cm3,(0.73±0.15)g/cm3比(0.80±0.16)g/cm3,t值分别为12.01、6.79、3.21、3.55、2.16,P值均<0.05]。研究组患者外周血INF-γ/IL-4与腰椎骨密度、髋骨骨密度均呈负相关性(r值分别为-0.78、-0.85,P值均<0.05)。Logistic回归分析显示,年龄≥60岁、吸烟、病程≥5年、II级以上关节功能、服用抗风湿性药物、INF-γ水平>5.51μg/L、IL-4水平≤0.45μg/L、INF-γ/IL-4>5.00是类风湿性关节炎继发骨质疏松症的危险因素(OR值分别为4.00、7.75、5.41、6.30、8.64、6.25、5.62、4.68,P值均<0.05);日饮牛奶、每周固定锻炼是类风湿性关节炎继发骨质疏松症的保护因素(OR值分别为0.57、0.50,P值均<0.05)。结论类风湿性关节炎患者的外周血INF-γ水平、INF-γ/IL-4较高,IL-4水平、骨密度较低,INF-γ/IL-4与骨密度呈负相关,INF-γ水平>5.51μg/L、IL-4水平≤0.45μg/L、INF-γ/IL-4>5.00与吸烟、服用抗风湿性药物均是继发骨质疏松症的危险因素。