Effect of Fugan Huaxian Decoction （扶肝化纤汤） on Th17/Treg Balance and Related Transcription Factors in Hepatic Fibrosis with Healthy Qi Deficiency and Toxin Accumulation Pattern Model Rats / 中医杂志

ObjectiveTo investigate the possible mechanism of Fugan Huaxian Decoction （扶肝化纤汤， FHD） against hepatic fibrosis （HF） from the perspective of immunity. MethodsForty-eight SD rats were randomly divided into blank group， model group， colchicine group， FHD high-， medium- and low-dose group， with eight rats in each group. Except for the blank group， the disease-syndrome combined model of HF with healthy qi deficiency and toxin accumulation pattern was established during six weeks in the other five groups. After successful modeling， the high-， medium- and low-dose FHD groups were respectively given 37.5， 18.75 and 9.38 g/（kg·d） of FHD granules by gavage， while the colchicine group received 2 mg/ （kg·d） of colchicine tablets by gavage， and the blank group and the model group were given 10 ml/（kg·d） of purified water， all for 3 weeks. The general condition of the rats was recorded. After the treatment， the histopathological morphology of the liver was observed by HE staining， and the levels of interleukin 10 （IL-10） and interleukin 17 （IL-17） in serum were determined by enzyme-linked immunosorbent assay （ELISA）. The expression of helper T cells 17 （Th17） and regulatory T cells （Treg） in peripheral blood were detected by flow cytometry， and the Th17/Treg value was calculated. The mRNA expression of retinoic acid-related nuclear orphan receptor γ （RORγt） and fork-head/wing-like helix transcription factor （FoxP3） in liver tissue were detected by qRT-PCR. ResultsCompared to the general condition of rats in the blank group， those in the model group were listless， less active， stretched and pushed， arched and prone， having no resistance to gavage， significantly reduced food intake， loose stools， dirty anus， slow weight gain， dry and dull hair， purple and darkening skin of the limbs with ecchymoses， purple and black spots with varying degrees of the skin of the tail； hepatic fibrosis and hyperplasia of rats in model group were more obvious； serum IL-17， peripheral blood Th17 expression and Th17/Treg value， RORγt mRNA expression in the liver tissue significantly increased in the model group， while expression of IL-10， Treg and FoxP3 mRNA significantly decreased （P＜0.05 or P＜0.01）. Compared to those in the model group， the general condition of the rats and the liver fibrosis of HE stained liver tissue were improved in all the medication groups； the expression of IL-17 and Th17， Th17/Treg， and RORγt mRNA expression significantly decreased， while expression of IL-10， Treg， and FoxP3 mRNA increased in the high- and medium-dose FHD groups and the colchicine group； the expression of IL-17， Th17， and RORγt mRNA decreased， while the expression of IL-10 and FoxP3 mRNA increased in the low-dose FHD group （P＜0.05 or P＜0.01）. And more improvements were found in the FHD high-dose group than FHD medium- and low-dose groups and colchicine group （P＜0.05 or P＜0.01）. ConclusionFHD can may regulate immune balance and act against fibrosis by regulating the expression of specific transcription factors FoxP3 and RORγt， affecting the differentiation of Th17 and Treg cells and Th17/Treg balance， and regulating the secretion of IL-10 and IL-17.

Absolute reduction in peripheral regulatory T cells in patients with Graves' disease and post-treatment recovery.

OBJECTIVE: Graves' disease (GD) is one of the most common autoimmune conditions, but the mechanisms underlying the associated induction of autoimmunity are not known. We explored the role of peripheral lymphocyte subpopulations in disease pathogenesis. METHODS: In total, 32 patients and 40 age- and sex-matched healthy controls were recruited in this study. Peripheral levels of T, B, NK, CD4+ T, CD8+ T, Th1, Th2, Th17, and Treg cells were measured using flow cytometry. For all patients, we compared all lymphocyte subpopulations between GD patients and healthy controls. Changes in patient lymphocyte subsets were compared before and after treatment. RESULTS: The absolute numbers of circulating Th17 cells (0.45 ± 1.16, p > 0.05) between GD patients and healthy controls were not significantly different. However, the percentage of Th17 cells was significantly increased (0.25 ± 0.11, p < 0.05). The absolute numbers and percentages of circulating Tregs in GD patients were significantly decreased compared with those in healthy participants (11.61 ± 2.75, p < 0.05). There was a significant difference in Treg absolute numbers between the untreated and drug-treated groups. Furthermore, we found that the Treg percentage in untreated patients (mean=4.78) was not significantly different from that in the drug-treated group (mean=4.81). In addition, circulating Treg absolute numbers in GD patients with exophthalmos were significantly lower than those in GD patients without exophthalmos (9.96 ± 4.16, p < 0.05). A similar trend was observed in GD patients with weight loss (11.97 ± 3.28, p < 0.05). CONCLUSION: GD pathogenesis was associated with a lower Treg population and an increased Th17/Treg ratio (T helper cell 17/ regulatory T cells). Th17 cells in this study were not related to the disease. Furthermore, anti-thyroid drug therapy improved immune-mediated system disorders. Finally, we found lower absolute numbers of circulating Tregs in GD patients with certain positive signs, such as exophthalmos and/or weight loss. Thus, immune changes are correlated with partial clinical manifestations.

Diagnostic Value of Bcl-2, miR-451 and Th17 Cells in Esophageal Cancer and Their Relation with Recurrence / 肿瘤防治研究

Objective To investigate the diagnostic value of Bcl-2, miR-451 and Th17 cells in esophageal cancer and their relation with recurrence. Methods We selected 101 patients with esophageal cancer as the experimental group and 95 healthy patients as the control group. The correlation between the clinicopathological characteristics and the level of each peripheral blood index was analyzed. The ROC curve was used to analyze the diagnostic value of each peripheral blood index. Multiple linear regression analysis was used to analyze the relation between each index and tumor recurrence. Results Peripheral blood Bcl-2, miR-451 and Th17 cells in the experimental group were higher than those in the control group (all P < 0.05); differentiation degree, clinical stage, tumor diameter and lymph node metastasis were positively correlated with the levels of Bcl-2, miR-451, and Th17 cells (all P < 0.05). Bcl-2, miR-451 and Th17 cells in relapsed patients were higher than those in non-relapsed patients (all P < 0.05); after controlling other factors such as differentiation degree, clinical stage, tumor size and lymph node metastasis, the levels of Bcl-2, miR-451 and Th17 cells were significantly correlated with recurrence (all P < 0.05). Conclusion Bcl-2, miR-451 and Th17 cells in peripheral blood of esophageal cancer patients are abnormally expressed and their expression are closely related to differentiation degree, clinical stage and recurrence. The combined detection could provide an objective basis for clinical diagnosis and prognosis assessment.

Research progress on the role of T helper cells 17 and interleukin 17 in oral mucosal diseases / 口腔疾病防治

@#T helper 17 (Th17) cells are a new type of CD4+ T helper cell. They participate in the immune and inflammatory response by secreting specific interleukin-17 (IL-17). In oral mucosal diseases, oral lichen planus (OLP), recurrent aphthous ulcer (RAU) and Behcet′s disease (BD) are associated with Th17 cells and IL-17. There were 17 kinds of proteins in the saliva of patients with OLP that could upregulate the expression of Th17 cells and induce the secretion of IL-17. IL-17 can stimulate epithelial cells, endothelial cells and fibroblasts to produce a variety of cytokines, such as IL-6, IL-8, granulocyte macrophage colony-stimulating factor and cell adhesion molecule-1, leading to the production and aggravation of inflammation. Th17/Tc17 cell-targeted therapy can significantly improve the clinical symptoms of OLP patients′ mucosa and skin. IL-17 can stimulate oral keratinocytes through the IL-17RA or IL-17RE receptor and produce proinflammatory effects in RAU. Th17 cells in the peripheral blood of BD patients are significantly increased, while Treg cells are significantly decreased.

[The effects and immunologic mechanisms of Helicobacter pylori infection on AOM/DSS induced colitis-associated cancer in mice].

Objective: To investigate the effects and potential mechanisms of Helicobacter pylori (H.pylori) infection on azoxymethane (AOM)/dextran sulfate sulphate (DSS) induced colitis-associated cancer (CAC) in mice. Methods: A total of 60 specific pathogen free C57BL/6J mice were randomly divided into four groups: normal control group (control group, n=9), H. pylori-infected group (Hp group, n=9), AOM/DSS-treated group (AOM/DSS group,n=21) and AOM/DSS-treated with H.pylori infection group (Hp+AOM/DSS group, n=21). Mice were sacrificed on day19, 45 or 85 after AOM/DSS challenge. Histopathological changes in colonic tissues were determined by hematoxylin and eosin staining. Flow cytometry analysis was performed to determine T helper cells 17 (Th17) and regulatory T cells (Treg) in colonic lamina propria. The expression levels of Th17-and Treg-associated cytokines and transcription factors [interleukin (IL)-10, IL-17A, retinoic acid receptor-related orphan receptor γt (RORγt) and forkhead box P3 (Foxp3)] were determined by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Results: There were no histopathological changes in colonic tissues of mice in control group and Hp group. H.pylori colonization reduced the histopathological scores at the stages of colitis (day 19) and dysplasia (day 45), and also decreased tumor load (day 85) in mice treated with AOM/DSS (all P<0.05). Compared with AOM/DSS group, the percentages of CD3(+)CD4(+)IL-17A(+)Th17 and CD3(+)CD4(+)IL-17A(+)Foxp3(+)Treg cells (1.88±0.17 vs 2.07±0.89, 1.06±0.13 vs 1.89±0.23) and the expression levels of RORγt and IL-17A (1.08±0.59 vs 2.35±1.35, 2.96±0.92 vs 7.78±4.57) were decreased in colonic tissues of Hp+AOM/DSS group (all P<0.05). The percentages of CD3(+)CD4(+)CD25(+)Foxp3(+)Treg and CD3(+)CD4(+)IL-10(+)Foxp3(+)Treg cells (20.60±3.39 vs 15.63±2.71, 2.94±0.52 vs 2.14±0.47) and the expression levels of Foxp3 and IL-10 [17.59(13.77,24.87) vs 6.27(4.41,13.36), 3.52(1.59,5.99) vs 1.17(1.15,2.75)] in colonic tissues were higher (all P<0.05) in mice of Hp+AOM/DSS group compared with AOM/DSS group on day 85. Conclusion: H.pylori infection slows the progress from inflammation to tumor in a AOM/DSS induced CAC modal, accompanied with the downregulation of Th17 response and upregulation of Treg response.

Progress of immune functions of T helper cells 17 and regulatory T cells and their immunomodulatory effect on non-small cell lung cancer / 肿瘤研究与临床

Under the regulation of various cytokines and different concentrations of cytokines, primary CD4+ T cells can differentiate into different Th subgroups. T helper cells 17 (Th17) and regulatory T cells (Treg cells) are research hotspots in recent years. They play diverse immunomodulatory roles by secreting various target cytokines. Th17 and Treg cells are different from each other but connect with each other, their regulatory mechanism is complex, and they play important roles in non-small cell lung cancer (NSCLC). This paper reviews the progress of the differentiation, the development and the immunological function of Th17 and Treg cells and their immunomodulatory effect on the NSCLC.

Effects of compound sophora flavescens injection on T helper cells 17, regulatory T cells and the related cytokines in lung cancer patients with cirrhosis / 肿瘤研究与临床

Objective@#To investigate the effect of compound sophora flavescens injection on T helper cells 17 (Th17), regulatory T-cells (Treg cells) and the related cytokines in lung cancer patients with cirrhosis.@*Methods@#A total of 63 patients with early non-small cell lung cancer and cirrhosis in Shouguang People's Hospital of Shandong Province from January 2016 to January 2018 were selected. All patients were scheduled to undergo surgical treatment. The patients were randomly divided into observation group (33 cases) and control group (30 cases) according to the random number table method. The patients in the control group were treated with liver protection, and the patients in the observation group were given compound sophora flavescens on the basis of the control group. After 3 months of treatment, the changes of liver fibrosis index, the levels of Th17 and Treg cells in peripheral blood and the changes of interleukin (IL)-10, IL-17, IL-6, and transforming growth factor (TGF)-β in serum were compared between the two groups.@*Results@#After treatment, the alanine aminotransferase (ALT) of 15 cases (45.45%) in the observation group were normalized and 6 cases (20.00%) in the control group were normalized. There was a significant difference in ALT normalization rate between the two groups (P < 0.05). The levels of serum laminin (LN) and hyaluronic acid (HA) in the two groups after treatment were lower than those before treatment (all P < 0.05), and the serum LN [(156.74±30.52) ng/ml] and HA [(179.56±25.32) ng/ml] in the observation group after treatment was lower than those in the control group [(210.58±39.42) ng/ml and (203.75±28.79) ng/ml] (t values were 18.236 and 12.184, both P < 0.01). There was no significant difference in the level changes of type Ⅲ procollagen and type Ⅳ collagen between the two groups before and after treatment (all P > 0.05). After treatment, the levels of Th17 and Th17/Treg increased and the level of Treg cells decreased (all P < 0.05). After treatment, the levels of Th17 [(1.32±0.18)%] and Th17/Treg (0.23±0.04) in the observation group were higher than those in the control group [(1.21±0.17)% and 0.20±0.03] (t values were 3.201 and 1.087, both P < 0.01), while the level of Treg cells in the observation group was lower than that in the control group (P < 0.05). After treatment, the serum levels of IL-10, IL-17, IL-6, and TGF-β in the two groups were lower than those before treatment (all P < 0.05), and their levels in the observation group were lower than those in the control group (all P < 0.05).@*Conclusion@#Compound sophora flavescens injection can improve the liver function and fibrosis index of patients with lung cancer and cirrhosis, and it can regulate the levels of Th17, Treg cells and their related factors, which can be used as a clinical adjuvant.

Metformin attenuates autoimmune disease of the neuromotor system in animal models of myasthenia gravis.

Metformin, the most widely used medicine for type 2 diabetes, displays anti-inflammatory functions via activating AMP-activated protein kinase (AMPK). Circulating autoantibodies and disequilibrium of helper T cells and regulatory T cells are pathological hallmarks of myasthenia gravis (MG). Rectify the imbalance of different T cell populations has become an important therapeutic strategy to treat MG. In this study, we assessed the effect of metformin on the development of autoimmunity using an experimental autoimmune myasthenia gravis (EAMG) rat model. We first provided evidence that oral administration of metformin attenuated the onset of EAMG. This effect was accompanied by a substantial decrease of circulating auto-antibody levels with no effect on blood glucose level. While metformin treatment in vitro showed little effect on inducible Treg, metformin strongly inhibited Th17 cell differentiation through the increase of reactive oxygen species and AMPK. Furthermore, an attenuation of antigen-induced IgG2b antibody production by two different doses of metformin was also observed in the AChR-specific recall response. In conclusion, the above results indicate that metformin may have therapeutic value for the clinical treatment of MG.

The effects of freeze-dried Ganoderma lucidum mycelia on a recurrent oral ulceration rat model.

BACKGROUND: Conventional scientific studies had supported the use of polysaccharides and ß-glucans from a number of fungi, including Ganoderma lucidum for the treatment of recurrent oral ulceration (ROU). Our aim of the present study was to evaluate whether freeze-dried powder from G. lucidum mycelia (FDPGLM) prevents ROU in rats. METHODS: A Sprague-Dawley (SD) rat model with ROU was established by autoantigen injection. The ROU rats were treated with three different dosages of FDPGLM and prednisone acetate (PA), and their effects were evaluated according to the clinical therapeutic evaluation indices of ROU. RESULTS: High-dose FDPGLM induced significantly prolonged total intervals and a reduction in the number of ulcers and ulcer areas, thereby indicating that the treatment was effective in preventing ROU. Enzyme-linked immunosorbent assay (ELISA) showed that high-dose FDPGLM significantly enhanced the serum transforming growth factor-ß1 (TGF-ß1) levels, whereas reduced those of interleukin-6 (IL-6) and interleukin-17 (IL-17). Flow cytometry (FCM) showed that the proportion of CD4+ CD25+ Foxp3+ (forkhead box P3) regulatory T cells (Tregs) significantly increased by 1.5-fold in the high-dose FDPGLM group compared to that in the rat model group (P < 0.01). The application of middle- and high-dose FDPGLM also resulted in the upregulation of Foxp3 and downregulation of retinoid-related orphan receptor gamma t(RORγt) mRNA. CONCLUSION: High-dose FDPGLM possibly plays a role in ROU by promoting CD4+ CD25+ Foxp3+ Treg and inhibiting T helper cell 17 differentiation. This study also shows that FDPGLM may be potentially used as a complementary and alternative medicine treatment scheme for ROU.