内镜下乳头括约肌小切开联合球囊扩张治疗胆管结石患者的疗效及对血清CCK-A、SREBP-2水平的影响

【目的】探讨内镜下乳头括约肌小切开联合球囊扩张治疗胆管结石的疗效及对血清胆囊收缩素A型受体(CCK-A)、胆固醇调节元件结合蛋白2(SREBP-2)水平的影响。【方法】选取2019年1月至2023年12月本院收治的196例胆管结石患者,根据随机数字表法将其分为观察组和对照组,每组98例。对照组患者采用内镜下乳头括约肌切开术,观察组患者行内镜下乳头括约肌小切开联合球囊扩张术。比较两组手术时间、出血量、住院时间、下床时间、胃肠功能恢复时间,比较两组取石率、结石复发率及术后并发症发生率。【结果】观察组手术时间、住院时间、下床时间、胃肠功能恢复时间短于对照组,出血量少于对照组,差异有统计学意义(P<0.05)。观察组一次取石成功率高于对照组(P<0.05),观察组机械碎石率、结石复发率低于对照组(P<0.05);观察组残余结石率与对照组比较,差异无统计学意义(P>0.05)。术后,两组患者CCK-A高于术前,SREBP-2低于术前;观察组术后CCK-A高于对照组,SREBP-2低于对照组,差异有统计学意义(P<0.05)。观察组术后并发症总发生率低于对照组,差异有统计学意义(P<0.05)。【结论】内镜下乳头括约肌小切开联合球囊扩张术治疗胆管结石具有手术时间短、术后恢复快、术后胆囊功能恢复好、并发症少等优势。

Homoharringtonine promotes heart allograft acceptance by enhancing regulatory T cells induction in a mouse model

Background:Homoharringtonine(HHT)is an effective anti-inflammatory,anti-viral,and anti-tumor protein synthesis inhibitor that has been applied clinically.Here,we explored the therapeutic effects of HHT in a mouse heart transplant model.Methods:Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver,kidney,and hematology.A mouse heart transplantation model was constructed,and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan-Meier analysis,immunostaining,and bulk RNA sequencing analysis.The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells(Tregs)differentiation.Results:HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo.Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days(P<0.001)without non-immune toxicity.The allografts had long-term survival after continuous HHT treatment for 28 days.HHT significantly reduced lymphocyte infiltration in the graft,and interferon-γ-secreting CD4^(+)and CD8^(+)T cells in the spleen(P<0.01).HHT significantly increased the number of peripheral Tregs(about 20%,P<0.001)and serum interleukin(IL)-10 levels.HHT downregulated the expression of T cell receptor(TCR)signaling pathway-related genes(CD4,H2-Eb1,TRAT1,and CD74)and upregulated the expression of IL-10 and transforming growth factor(TGF)-βpathway-related genes and Treg signature genes(CTLA4,Foxp3,CD74,and ICOS).HHT increased CD4^(+)Foxp3^(+)cells and Foxp3 expression ex vivo,and it enhanced the inhibitory function of inducible Tregs.Conclusions:HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels,thereby promoting mouse heart allograft acceptance.These findings may have therapeutic implications for organ transplant recipients,particularly those with viral infections and malignancies,which require a more suitable anti-rejection medication.

Early changes of CD4＋CD25＋Foxp3＋ regulatory T cells and Th1/Th2, Tc1/Tc2 profiles in the peripheral blood of rats with controlled hemorrhagic shock and no fluid resuscitation

Background Hemorrhagic shock induces immune dysfunction. Regulatory T cells （Tregs）, T-helper （Th） cells, and cytotoxic T-lymphocytes （CTLs） can execute many crucial actions in immune and inflammatory responses. This study was conducted to investigate the early pathophysiological changes of CD4＋CD25＋Foxp3＋ Treg and Th1/Th2, Tc1/Tc2 profiles in the peripheral blood of rats with controlled hemorrhagic shock and no fluid resuscitation. Methods A rat model of controlled hemorrhagic shock with no fluid resuscitation was established. Peripheral blood samples were taken before and four hours after hemorrhagic shock with no fluid resuscitation. Three color flow cytometry was used to detect Tregs, Thl, Th2, Tcl and Tc2 ceils in the samples. Results In the peripheral blood of rats, the percentage of Tregs four hours after hemorrhagic shock was significantly lower than before hemorrhagic shock （P=0.001）. The ratios of Th1/Th2 and Tc1/Tc2 were changed from （23.08±8.98）% to （23.91±15.36）%, and from （40.40±21.56）% to （65.48±23.88）%, respectively. Conclusions At an early stage, the advent of hemorrhagic shock is related to an early decrease of Tregs, and a mild shift in the Th1/Th2, Tc1/Tc2 balance toward Thl and Tcl dominance. These changes are part of a hyper-inflammatory state of the host, and will deteriorate the maintenance of immune balance. Further influences and detailed mechanisms need to be investigated.

不同分离方法对小鼠脾脏调节性T细胞表达特征的影响

近年来,随着以变应性鼻炎（AR）和变应性哮喘为代表的呼吸道变应性疾病在全球范围内发病率的升高,与之发病机制密切相关的CD4＋CD25＋T细胞及其特异性标志物Foxp3成为近年来的研究热点。在此前的实验中,我们对AR小鼠外周调节性T细胞（regulatory T cell,Treg）的表达特征进行了研究,发现实验组与正常对照组外周血表达有显著性差异,而脾脏表达两组间无显著性差异[1]。

Effects of lysedEnterococcus faecalis FK-23 on experimental allergic rhinitis in a murine model

In the current study, we sought to investigate whether lysed Enterococcus faecalis FK-23 （LFK）, a heat-killed probiotic preparation, attenuated eosinophil influx into the upper airway and had immunomodulatory activity in a murine allergic rhinitis model. Eighteen BALB/c mice were divided into three groups; the ovalbumin （OVA）-sen- sitized/challenged group, which received saline orally for 6 weeks （OVA group）, the OVA-sensitized/challenged group, which received LFK orally for 6 weeks （LFK-fed group）, and the non-sensitized group, which received saline for 6 weeks （saline control group）. Nasal rubbing and sneezing were monitored during the study. After the final challenge, interleukin （IL）-4, interferon （IFN）-y, and OVA-specific IgE levels in the sera and splenocyte culture supernatants were determined, eosinophilic infiltrate into the upper airway was quantified, and splenic CD4~CD25＋ regulatory T cells （Tregs） were examined by flow cytometry. We found that nasal rubbing was sig- nificantly reduced in LFK-fed mice compared to the OVA group on d 27 and 35, and sneezing was significantly inhibited by LFK administration for 35 d. LFK-fed mice had significantly less eosinophil influx into the nasal mucosa than the OVA group. There were no significant differences between the LFK-fed group and OVA group in the serum and splenocyte culture supernatant levels of IL-4, IFN-y, and OVA-specific IgE. Interestingly, the LFK-fed mice had a significantly greater percentage of splenic CD4＋CD25＋ Tregs than OVA group. Our results indicate that oral administration of LFK may alleviate nasal symptoms, reduce nasal eosinophilia, and increase the percentage of CD4＋CD25＋ Tregs in experimental allergic rhinitis.

Modified Shenlingbaizhu decoction reduces intestinal adenoma formation in adenomatous polyposis coli multiple intestinal neoplasia mice by suppression of hypoxia-inducible factor 1α-induced CD4+CD25+forkhead box P3 regulatory T cells

OBJECTIVE： To test the hypothesis that modified Shenlingbaizhu decoction （MSD） attenuates the for- mation of intestinal adenomas by regulating activa- tion of CD4＋CD25＋ forkhead box P3 （FoxP3） regu- latory T cells （Tregs） by downregulation of hypox- ia-inducible factor la （HIF-la）. METHODS： Chemical fingerprints of ginsenoside Rbl, ginsenoside Rc, paeoniflorin, and dioscin in standard extractions were used as material bases of MSD. Adenomatous polyposis coli multiple intesti- nal neoplasia （ApcM＇n/＋） mice, which harbor a muta- tion in adenomatous polyposis coil, were used to host intestinal adenomas. Peripheral blood and spleen Tregs were analyzed by flow cytometry. Pro- tein expression was analyzed by immunohisto- chemistry and Western blotting. RESULTS： The number and size of intestinal adenomas were significantly reduced by MSD treatment. Mucosal thickening and the spleen size were also substantially decreased by MSD. The carcinogenesis process in Apc^min/＋ mice resembled that of human colorectal cancer. Molecular markers of neoplasms, such as 13-catenin, cyclooxygenase-2, prolif- erating cell nuclear antigen, and p53, were substantially ameliorated by MSD treatment. Moreover, MSD downregulated peripheral and spleen CD4＋ CD25＋FoxP3＋ Tregs and reduced in situ expression of CD4, CD25, and FoxP3 in intestinal adenomas. MSD also suppressed HIF-la expression in the intestinal adenomas, and HIF-la inhibition decreased expression of FoxP3 in Jurkat T cells under hypoxic conditions. CONCLUSION： MSD is a valid prescription to control the formation of intestinal adenomas in Apc^min/＋mice. It exerts anti-cancer effects partially through suppression of HIF-la that induced activation of CD4＋CD25＋FoxP3＋ Tregs in vivo and in vitro.

Regulatory T cells and cardiovascular diseases

Inflammation is a major underlying mechanism in the progression of numerous cardiovascular diseases(CVDs).Regulatory T cells(Tregs)are typical immune regulatory cells with recognized immunosuppressive properties.Despite the immunosuppressive properties,researchers have acknowledged the significance of Tregs in maintaining tissue homeostasis and facilitating repair/regeneration.Previous studies unveiled the heterogeneity of Tregs in the heart and aorta,which expanded in CVDs with unique transcriptional phenotypes and reparative/regenerative function.This review briefly summarizes the functional principles of Tregs,also including the synergistic effect of Tregs and other immune cells in CVDs.We discriminate the roles and therapeutic potential of Tregs in CVDs such as atherosclerosis,hypertension,abdominal arterial aneurysm,pulmonary arterial hypertension,Kawasaki disease,myocarditis,myocardial infarction,and heart failure.Tregs not only exert anti-inflammatory effects but also actively promote myocardial regeneration and vascular repair,maintaining the stability of the local microenvironment.Given that the specific mechanism of Tregs functioning in CVDs remains unclear,we reviewed previous clinical and basic studies and the latest findings on the function and mechanism of Tregs in CVDs.

Effect of partial depletion of CD25+ T cells on neurological deficit and tissue damage in acute cerebral ischemia rat models

Objective: To evaluate the role of regulatory T cells (Tregs) at late stages of stroke. Methods:Anti-CD25 antibody (or PBS as a control) was injected to reduce the pool of Tregs in Wistar rats;then, ischemia was induced transiently by middle cerebral artery occlusion during 60 min and reperfusion was allowed for 7 d. Then, Treg frequency was analyzed in peripheral blood, spleen and lymph nodes. Neurological score (0-6) and infarct volume were also determined. Results: Nine days after injection, the CD4+CD25+ T cells were reduced by 70.4%, 44.8% and 57.9% in peripheral blood, spleen and lymph nodes, respectively compared to PBS-treated rats. In contrast, the reduction of CD4+FOXP3+ T cells was lower in the same compartments (38.6%, 12.5%, and 29.5%, respectively). The strongest reduction of CD25+CD4+ T cells was observed in those FOXP3-negative cells in blood, spleen and lymph nodes (77.8%, 52.8%, and 60.7%, respectively), most likely corresponding to activated T cells. Anti-CD25-treated transient middle cerebral artery occlusion rats had a lower neurological deficit and did not develop tissue damage compared with PBS-treated animals. Conclusions: These findings suggest that treatment with anti-CD25 in our model preferentially reduce the T cell population with an activated phenotype, rather than the Treg population, leading to neuroprotection by suppressing the pathogenic response of effector T cells.

伴绝经后骨质疏松症的股骨颈骨折患者外周血Treg细胞频率、Treg细胞相关因子表达水平的变化及其临床意义

【目的】分析伴绝经后骨质疏松症(PMOP)的股骨颈骨折患者外周血Treg细胞频率、Treg细胞相关因子表达水平的变化及其临床意义。【方法】选择2016年10月至2020年9月长沙市中心医院收治的30例伴PMOP的股骨颈骨折患者(伴PMOP骨折组)和30例无PMOP的股骨颈骨折女性患者(无PMOP骨折组),并选择同期健康体检的30例无骨折的健康绝经后女性志愿者(对照组)。采用化学发光法检测血清骨钙素(BGP)、Ⅰ型胶原羧基端肽(b-CTX)、Ⅰ型胶原N-端前肽(PINP)水平,采用流式细胞学技术检测各组Treg细胞频率,采用ELISA检测白细胞介素10(IL-10)、转化生长因子β(TGF-β)水平。比较三组研究对象Treg细胞频率、IL-10、TGF-b、BGP、b-CTX、PINP水平。【结果】伴PMOP骨折组患者Treg细胞频率及血清IL-10、TGF-β水平低于无PMOP骨折组患者及对照组(P<0.05),对照组与无PMOP骨折组患者组间比较,差异无统计学意义(P>0.05)。伴PMOP骨折组患者血清b-CTX、PINP水平高于无PMOP骨折组患者及对照组,BGP水平低于无PMOP骨折组患者及对照组,差异均有统计学意义(P<0.05)。Pearson相关性分析显示,血清b-CTX、PINP水平与Treg细胞频率均呈负相关(r分别为-0.262、-0.255,均P<0.05)。【结论】伴PMOP的股骨颈骨折患者Treg细胞频率降低,Treg细胞相关因子表达水平偏低,Treg细胞数目减少可能与PMOP发病有关。

miR-155 HG在寻常型银屑病患者外周血单个核细胞中的表达水平及其与Th17/treg细胞平衡的关系

【目的】探讨microRNA-155宿主基因(miR-155 HG)在寻常型银屑病患者外周血单个核细胞(PBMC)中的表达水平及其与辅助性T细胞17/调节性T淋巴细胞(Th17/treg)细胞平衡的关系。【方法】选择2020年10月至2021年10月在榆林市第二医院诊治的寻常型银屑病患者92例血液标本为疾病组,另外选取同期健康体检者92例血液标本为对照组,采用实时定量PCR(RT-PCR)检测PBMC中miR-155 HG、miR-155表达水平,采用流式细胞仪检测Th17、treg细胞比例,计算Th17/treg值;采用酶联免疫吸附试验(ELISA)检测血清白细胞介素-6(IL-6)、转化生长因子β1(TGF-β1)水平;采用Pearson法分析PBMC中miR-155 HG、miR-155水平及其与Th17/treg值、IL-6、TGF-β1水平的相关性。【结果】与对照组比较,疾病组患者PMBC中miR-155 HG、miR-155、IL-6水平及Th17/treg值均显著增加(P<0.05),血清TGF-β1水平降低(P<0.05)。疾病组患者PBMC中miR-155 HG水平与miR-155水平呈正相关(r=0.573,P=0.000);miR-155 HG水平与Th17/treg值、IL-6水平呈正相关(r=0.482、0.512,P=0.000),与TGF-β1水平呈负相关(r=-0.596,P=0.000);miR-155水平与Th17/treg值、IL-6水平呈正相关(r=0.694、0.488,P=0.000),与TGF-β1水平呈负相关(r=-0.509,P=0.000)。【结论】寻常型银屑病患者PBMC中miR-155 HG水平上调,可能与分泌miR-155、Th17/treg细胞失衡有关。

Effect of Jiaweiwumei decoction on regulatory T cells and interleukin-10 in a rat model of ulcerative colitis

OBJECTIVE:To investigate the effect of a decoction made from the Traditional Chinese Medicine Wumei pill,on regulatory T cells and interleukin-10(IL-10) in a rat model of ulcerative colitis induced by2,4,6-trinitrobenzene sulfonic acid(TNBS).METHODS:Rat ulcerative colitis was induced with TNBS.All modeled rats were randomly divided into six groups:normal control group;model group;sulfasalazine suppositories treatment group;and high,moderate,and low dosage of Jiaweiwumei decoction groups(12 rats each).Colon injury index was evaluated after 14 days.After peripheral blood lymphocyte separation,CD4 + T cells and CD4+/CD25+ T cell percentage was detected by flow cytometry.The content of IL-10 in serum and intestinal mucosa tissue was detected by sandwich enzyme-linked immunosorbent assay.RESULTS:Colon injury indices in the decoction groups were effectively reduced,compared with the model group(P < 0.05).Compared with that of the control group,the CD4+/CD25+ to CD4+ T lymphocyte ratio of the model group was significantly lower,while the decoction treatment improved the CD4 +/CD25 + to CD4 + T lymphocyte ratio(P <0.05).The serum and mucosal IL-10 content of the model group was significantly lower(P < 0.05) than that in the control group,while the decoction group had significantly higher serum and intestinal mucosal IL-10 content than that in the model group(P < 0.05).The regulatory T cell content was negatively correlated with the colonic injury index(r = 0.68,P < 0.05),and positively correlated with the content of serum IL-10(r = 0.87,P < 0.05) and intestinal mucosal IL-10(r= 0.79,P < 0.05).CONCLUSION:Jiaweiwumei decoction had significant effects on regulatory T cells and IL-10 in rats with TNBS-induced ulcerative colitis.

Bupi Hewei decoction ameliorates 5-fluorouracil-induced intestinal dysbiosis in rats through T helper 17/T regulatory cell signaling pathway

OBJECTIVE:To observe the effects of the Bupi Hewei(BPHW)decoction on diarrhea and intestinal flora disorder induced by 5-fluorouracil(5-FU)and investigate the possible mechanism underlying these actions.METHODS:Thirty-five male Sprague-Dawley rats were randomly divided into four groups:normal control,5-FU,5-FU+BPHW decoction(10.5 g/kg for 5 consecutive days),and 5-FU+Bacillus licheni-formis capsule groups(0.2 g/kg for 5 consecutive days).Animal models were established via the intraperitoneal injection of 5-FU(30 mg/kg for 5 consecutive days).At the end of the treatment period,diarrhea was assessed,and the change of the intestinal flora was examined using 16 S r DNA highthroughput sequencing.Interleukin(IL)-17,IL-21,IL-6,IL-10,RAR-related orphan receptor gamma(RORγt),and forkhead box P3(Foxp3)expression in the jejunum was detected using immunohistochemistry,quantitative real-time polymerase chain reaction(PCR),Western blotting,and enzymelinked immuno sorbent assay.RESULTS:In this study,the BPHW decoction effectively lowered the diarrhea score,increased the proportions of Bacteroidetes and Prevotellaceae-Alloprevotella species,and reduced the proportions of Proteobacteria,Escherichia-Shigella,Ruminococcaceae NK4 A214,and Ruminococcaceae UCG-005 species in the rat intestine after 5-FU chemotherapy.In addition,the BPHW decoction significantly suppressed the expression of IL-17,IL-21,IL-6,IL-10,RORγt,and Foxp3 in the jejunum.CONCLUSION:Our findings suggest that the BPHW decoction can improve the intestinal immune balance and reduce intestinal inflammation by targeting T helper cell/T regulatory cell-associated factors.

HLA-DR expression on regulatory T cells is closely associated with the global immune activation in HIV-1 infected subjects naive to antiretroviral therapy

Background The frequencies of regulatory T cells （Tregs） increased over the HIV infection but its counts actually decreased. We proposed that the decrease of Treg counts may cause the reduction of inhibitory effect and thereby account for the over-activation of Tregs during HIV infection. However, it remains unknown whether Tregs are also over-activated and thereafter the activation induced death may lead to the decrease of Tregs. Methods Tregs were defined as CD4＋CD25＋CD127lo/-T cells. Eighty-one HIV-1 infected patients were enrolled in our study, and twenty-two HIV-1 seronegative donors were recruited as the control. The levels of HLA-DR on Tregs were determined by FACSAria flow cytometer. Results Compared to HIV-1 seronegative donors, the levels of HLA-DR on CD4＋CD25＋CD127lo/- Tregs were significantly increased in HIV-1 infected patients, and its increase was positively associated with viral loads （r=0.3163, P=-0.004） and negatively with CD4 T-cell counts （r=-0.4153, P 〈0.0001）. In addition, significant associations between HLA-DR expression on CD4＋CD25＋CD127lo/- Tregs and the percentages of HLA-DR, CD38, Ki67 expressing CD4＋ and CD8＋ T cells were also identified. Conclusion HLA-DR on Tregs is a good marker for viral replication and disease progression. The over-activation of Tregs might result in the decrease of Tregs.

Efficacy of Wumei Baijiang prescription(乌梅败酱方)on regulatory T cells/helper T cells Immune balance in mice with ulcerative colitis

OBJECTIVE:To observe the therapeutic effect of Wumei Baijiang prescription(乌梅败酱方),empirical prescription of Lu Zhizheng,on experimental ulcerative colitis(UC)mice,and to investigate the mechanism of the prescription in UC from the perspective of the immune balance of regulatory T cells(Treg)and helper T cells(Th17).METHODS:Sixty C57 BL/6 mice were randomly divided into 6 groups:normal group,model group,Chinese medicine group(high,medium and low dose group of Wumei Baijiang prescription)and control group(mesalazine sustained-release granules).Except for the normal group,the other groups used 2.5%dextran sulfate sodium to induce UC mice model.At the end of the model,the Chinese medicine group was given high,medium and low dose administration of Wumei Baijiang prescription,the control group was given slow-release granules of mesalazine,and the model group was given equal volume saline for 10 d.The changes of food intake,body weight,disease activity index(DAI)score,length of large intestine and histopathology were observed.The number of Treg,Th17,CD4+,CD8+cells in spleen was detected by flow cytometry.The expression of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10)and C-reactive protein(CRP)in serum was detected by enzyme-linked immunosorbent assay.RESULTS:The middle and high-dose groups of Wumei Baijiang prescription were superior to the model group in terms of increasing food intake and body weight of colitis mice,restoring colon morphology,improving pathological damage,and reducing DAI(P<0.05).There was no statistical difference with the mesalazine group(P>0.05).Compared with the model group,the spleen Treg and CD4+of the mice in the high and middle dose groups of Wumei Baijiang prescription were higher,while Th17 and CD8+were lower(P<0.05),and there was no statistical difference compared with the mesalazine group(P>0.05).In addition,compared with the model group,the serum levels of TNF-αand CRP in mice with high and middle doses of Wumei Baijiang prescription and mesalazine group were lower(P<0.05),and IL-10 content was higher(P<0.05).CONCLUSIONS:Wumei Baijiang prescription can improve the general conditions of colitis mice,such as diarrhea,hematochezia,weight loss,and mucosal damage.The mechanism may be related to the regulation of Treg/Th17 immune balance.

天灸“肺俞”穴对健康大鼠Th17/Treg免疫平衡及炎症反应的影响

目的探讨天灸“肺俞”穴对健康大鼠辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将48只大鼠随机分为假天灸组10只,天灸组38只。假天灸组大鼠将空白的穴位贴贴敷于双侧“肺俞”穴,天灸组大鼠将填有复方斑蝥膏的穴位贴贴敷于双侧“肺俞”穴,贴敷时间为8 h。8 h后天灸组大鼠成功发泡30只,再随机分为天灸1天组、天灸3天组、天灸7天组,每组10只。实验期间记录大鼠一般情况;天灸不同时间组在相应时间取材,采用HE染色观察大鼠贴敷区域皮肤的病理改变;采用流式细胞术检测外周血Th17、Treg细胞表达,计算Th17/Treg值;采用ELISA法检测血清白细胞介素17A(IL-17A)、白细胞介素6(IL-6)、白细胞介素10(IL-10)含量。结果与假天灸组比较,天灸1天组大鼠贴敷区域可见水泡,大鼠经常搔抓天灸区域皮肤,皮肤出现角质层松散,棘层、颗粒层增厚,基底层细胞缺失,炎性细胞浸润,外周血Treg及血清IL-17A、IL-6水平升高(P<0.01);天灸3天组天灸区域结痂并可见部分脱落,皮肤病理改变最明显,外周血Th17、Treg表达及血清IL-17A、IL-6、IL-10水平升高(P<0.01);天灸7天组天灸区域皮肤损伤及病理改变基本恢复。天灸1、3、7天组Th17/Treg值均较假天灸组降低(P<0.05或P<0.01)。与天灸1天组比较,天灸3天组外周血Th17、Treg表达及血清IL-17A升高;天灸7天组外周血Treg表达及血清IL-17A、IL-6降低,Th17/Treg值升高(P<0.05或P<0.01)。与天灸3天组比较,天灸7天组外周血Th17、Treg表达及血清IL-17A、IL-6、IL-10降低,Th17/Treg值升高(P<0.05或P<0.01)。结论天灸“肺俞”穴能使健康大鼠Th17/Treg平衡向Treg偏移,使其逐渐趋向免疫平衡,并调节血清中相关的炎症因子防止炎症的发生发展。

Effects of Xinfeng capsule on pulmonary function based on treg-mediated notch pathway in a rat model of adjuvant arthritis

OBJECTIVE:To observe the impact of xinfeng xapsule(XFC) on pulmonary function in a rat model of adjuvant arthritis(AA) and to investigate the mechanism of action.METHODS:Forty rats were randomly divided into four groups of ten:normal control(NC);model control(MC);tripterygium glycosides tablet(TPT);and xinfeng capsule(XFC).Except for the NC group,AA was induced in all rats by intracutaneous injection of 0.1 mL Freund's complete adjuvant in the right paw on the 19th day.NC and MC groups were given(0.9%) physiological saline.The TPT and XFC groups were given TPT(10 mg/kg) and XFC(1.2 g/kg),respectively.Thirty days after administration,changes in paw edema(E),the arthritis index(AI),pulmonary function,levels of regulatory T-cells(Treg),ultrastructure of lung tissue,and expression of Notch receptors and ligands in lung tissue were observed.RESULTS:In the MC group,E and the AI were increased and pulmonary function significantly decreased;the structure of alveolar type-II cells was damaged;ratios of Treg in peripheral blood were reduced;and expression of Notch receptors such as Notch3 and Notch4 and ligands such as Delta1 in lung tissue were significantly increased whereas expression of Notch1,Jagged1 and Jagged2 were significantly decreased.After intervention with XFC,E and the AI were decreased;pulmonary function was enhanced;the structure of alveolar type-II cells was improved;and expression of Treg,Notch1,Jagged1,Jagged2 was elevated,whereas that of Notch3,Notch4 and Delta1 was reduced.CONCLUSION:XFC can not only inhibit E and the AI and improve joint symptoms,it can also improve pulmonary function and reduce inflammation in lung tissue.These actions could be carried out through increases in the expression of Treg,Notch receptors(Notch1) and ligands(Jagged1,Jagged2),and reductions in the expression of Notch3,Notch4 and Delta1.These phenomena would reduce the deposition of immune complexes and the inflammatory response in lung tissue,thereby improving joint symptoms and pulmonary function.

Protective effects of Jiayan Kangtai granules on autoimmune thyroiditis in a rat model by modulating Th17/Treg cell balance

OBJECTIVE: To investigate the protective effects of Jiayan Kangtai(JYKT) granules, consisting of 9 Chinese herbs, in a rat model of autoimmune thyroiditis(AIT), and the possible underlying mechanism.METHODS: Female Lewis rats(6-8 weeks) were randomly apportioned to 5 groups of 10, including a normal control. AIT was induced in the untreated AIT-model group, and rats treated subsequently with daily low, medium, or high dose JYKT granules. After 12 weeks, plasma levels of thyroid autoantibodies and morphological changes in the thyroid were detected by enzyme-linked immunosorbent assay and histological examination, respectively. The presence of interleukin(IL)-6,IL23 p19, and IL-2 in thyroid tissue was assessed by immunohistochemical staining. The percentages of T helper(Th)17 cells and regulatory T cells(Tregs) in the peripheral blood were analyzed by flow cytometry. Relevant levels of cytokines and proteins were examined via bead-based multiplex flow cytometry and ELISA, respectively. Expressions of genes and proteins regulated by Th17 cells and Tregs were shown by real-time PCR and Western blot.RESULTS: Compared to the control, AIT-model rats had higher plasma concentrations of thyroid autoantibodies. The high-dose JYKT rats showed significantly lower levels of thyroid autoantibodies compared with the AIT model group. Rats in the AIT-JYKT groups also had fewer thyroid lesions and less lymphocytic infiltration, a lower percentage of Th17 cells, and a higher percentage of Tregs, compared with the AIT-model. Rats given high-dose JYKT had a significantly lower Th17/Treg ratio compared with the AIT model. Differences in plasma cytokine concentrations and relevant gene and protein expressions in the spleens of JYKT-treated rats and the AIT group suggested an association between JYKT treatment and lower Th17 cell percentage and higher Treg activity.CONCLUSION: JYKT treatment appeared to be protective against AIT in rats, possibly via the regulation of the Th17 cell/Treg imbalance in AIT.

Correlation analysis of Treg/Th17 cells and related cytokines in patients with psoriasis vulgaris

OBJECTIVE: To investigate potential differences in circulating levels of T regulatory (Treg)/T helper 17 (Th17) cells, related inflammatory cytokines and specific transcription factors in healthy individuals and patients with psoriasis conforming to one of three Traditional Chinese Medicine (TCM) syndromes: blood-heat syndrome (BHS), blood-stasis syndrome (BSS) and blood-dryness syndrome (BDS). METHODS: Sixty-seven patients with psoriasis were recruited and assigned to one of three corresponding TCM syndrome groups: BHS (n = 40), BSS (n = 14) and BDS (n = 13 patients). The control group comprised 21 healthy individuals. The circulating levels of Treg/Th17 cells in peripheral blood were assessed using flow cytometry;the levels of inflammatory cytokines interleukin (IL)-10 and tumor necrosis factor (TNF)-α by enzyme-linked immunosorbent assay;and the mRNA expression of T cell-specific transcription factors retinoic acid-related orphan receptor γt (RORγt) and forkhead box P3 (Foxp3) by quantitative real-time PCR. RESULTS: The ratio of Th17 cells and the levels of TNF-α and RORγt were all significantly higher in the BHS and BSS groups than the control group (P < 0.05), while the ratio of Treg cells and the levels of IL-10 and Foxp3 mRNA in the BHS group were significantly lower compared with the control group (P < 0.05). No significant differences were seen between the BSS group and the control group. The ratio of Th17 cells and the levels of TNF-α and RORγt in the BDS group were not significantly different from those of the control group;however, the ratio of Treg cells and the levels of IL-10 and Foxp3 were all lower than those in the healthy controls (P < 0.05). CONCLUSION: Compared with healthy individuals, the ratio of Th17 cells and the levels of related cytokines were higher, while the ratio of Treg cells and the levels of related cytokines were lower, in the peripheral blood of psoriasis/BHS patients;corresponding results for the BSS and BDS groups also showed differences. We propose that patterns of differentiation of immunological cells in psoriasis patients are reflected in corresponding TCM blood syndromes.

Role of interleukin-10 and transforming growth factor beta 1 in otitis media with effusion

Background Otitis media with effusion （OME） is a disease with complicated pathogeneses which are not clearly known Increasing interest has been focused on immunological cells, cytokines and their roles in chronic inflammatory states. This study was designed to disclose the existence and roles of interleukin-10 （IL-10） and transforming growth factor beta1 （TGF-β1） in the cause of OME in adults, and to investigate the probable role of Foxp3^＋CD4^＋CD25^＋ T cells in OME. Methods The concentrations of IL-10 and TGF-β1 in the middle ear effusions （MEEs） and plasmas of 36 adults （45 ears） with OME were measured by means of enzyme linked immunosorbent assay （ELISA）. As contrast, the concentrations of IL-10 and TGF-131 in the plasma of 30 normal volunteers were measured using the same method. Furthermore, the proportion of Foxp3^＋CD4^＋CD25^＋ T cells in CD4^＋ T cells of blood was tested by flow cytometry. Results （1） The concentrations of IL-10 in all MEEs and plasmas of the chronic OME patients were higher than those in patients with acute OME （both P 〈0.05）, so was TGF-131 （both P 〈0.01）. The concentration of IL-10 in MEEs was significantly higher than that in plasmas, not only in acute OME （P〈0.01）, but also in chronic OME （P〈0.01）. In chronic OME, the concentration of TGF-β1 in MEEs had no statistical difference with those in plasmas of the same patients. However, the concentration of TGF-β1 in plasmas of patients with chronic OME was significantly higher than that in plasmas of normal volunteers （P 〈0.01）. （2） The concentrations of IL-10 and TGF-β1 in MEEs of the patients who had been treated more than once were higher than those MEEs of the patients who were treated for the first time, respectively （P〈0.05, P〈0.01）. The level of TGF-β1 in plasmas of the patients who had been treated more than once was higher than in those of the patients who were treated firstly （P 〈0.05）, while the level of IL-10 in plasmas had no difference. The concentration of IL-10 in mucoid MEEs was higher than those in serous ones （P〈0.05）, while TGF-β1 had no statistical difference between mucoid and serous MEEs （P〉0.05）. The concentration of IL-10 in MEEs had a strong correlation with the duration of the illness （r=0.547, P〈0.01）. The same correlation was also found between the concentration of TGF-β1 in MEEs and the times patients being treated （r=0.579, P 〈0.01）. （3） The proportion of Foxp3^＋CD4^＋CD25^＋T/CD4^＋ T cells in the blood of chronic OME was not only significantly higher than that in the acute OME （P〈0.01）, but also higher than that in normal volunteers （P 〈0.01）. In chronic OME, there was a correlation between the proportion of Foxp3^＋CD4^＋CD25^＋ T/CD4^＋ T cells in the blood and the concentration of IL-10 in the plasmas （r=0.602, P 〈0.05）. Conclusions IL-10 and TGF-β1, as two important immunoregulatory mediators, participate in middle ear inflammatory response, especially in chronic course of OME in adults. Foxp3^＋CD4^＋CD25^＋ T cells may play some immunoregulatory roles in the course of this disease.

Immunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell lineImmunomodulatory effects of astragalus polysaccharide on Human peripheral blood mononuclear cells co-cultured with cervical cancer cell line

OBJECTIVE:To investigate immunomodulatory effects of Astragalus polysaccharides(APS)on the co-culture of peripheral blood mononuclear cells(PBMCs)with He La cervical cancer cell line.METHODS:To assess the proliferation of PBMCs,carboxyfluorescein succinimidyl ester(CFSE)-labeled PBMCs were co-cultured with He La cells and treated with different concentrations of APS.Supernatants of cell culture were collected for cytokines assay via enzyme-linked immunosorbent assay(ELISA).The impact of APS on the proliferation of PBMCs,induction of regulatory T cells(Tregs),and myeloid-derived suppressor cells(MDSCs)was carried out by flow cytometry.RESULTS:It was observed that APS could increase the proliferation of PBMCs co-cultured with He La cells(P<0.05).However,APS had no significant effects on the induction of Tregs and MDSCs in the co-culture assay(P>0.05).Furthermore,ELISA results demonstrated that APS could decrease IL-10 and TGF-βconcentration(P<0.05).CONCLUSION:The above-mentioned characteristics showed that APS might be able to modulate immune responses and improve anti-tumor effects through increasing the proliferation of PBMCs and decreasing inhibitory cytokines secretion as critical mediators of immune suppression in the tumor microenvironment.