Mangiferin corrects the imbalance of Th17/Treg cells in mice with TNBS-induced colitis

In the previous study, 80% ethanol extract of the rhizome mixture of Anemarrhena asphodeloides and Coptidis chinensis (AC) and its main constituent mangiferin improved TNBS-induced colitis in mice by inhibiting macrophage activation related to the innate immunity. In the preliminary study, we found that AC could inhibit Th17 cell differentiation in mice with TNBS-induced colitis. Therefore, we investigated whether AC and it main constituent mangiferin are capable of inhibiting inflammation by regulating T cell differentiation related to the adaptive immunity in vitro and in vivo. AC and mangiferin potently suppressed colon shortening and myeloperoxidase activity in mice with TNBS-induced colitis. They also suppressed TNBS-induced Th17 cell differentiation and IL-17 expression, but increased TNBS-suppressed Treg cell differentiation and IL-10 expression. Moreover, AC and mangiferin strongly inhibited the expression of TNF-α and IL-17, as well as the activation of NF-κB. Furthermore, mangiferin potently inhibited the differentiation of splenocytes into Th7 cells and increased the differentiation into Treg cells in vitro. Mangiferin also inhibited RORγt and IL-17 expression and STAT3 activation in splenocytes and induced Foxp3 and IL-10 expression and STAT5 activation. Based on these findings, mangiferin may ameliorate colitis by the restoration of disturbed Th17/Treg cells and inhibition of macrophage activation.     

维拉帕米对Th17/Treg细胞因子的效应研究

目的:研究维拉帕米(VER)对Th17/Treg细胞因子的效应作用。方法:用酶联免疫吸附试验(ELASA)法检测VER对脂多糖(LPS)激活的体外培养的人外周血淋巴细胞分泌Th17/Treg细胞因子的影响,观察VER对淋巴细胞增殖的效应。结果:正常儿童外周血淋巴细胞经脂多糖(LPS)激活后IL-17、IL-6和TGF-β1含量较未激活组显著增加(P<0.05);经LPS与10μmol.L-1VER共培养后,IL-17、IL-6和TGF-β1含量与未激活组比较无显著差异(P>0.05),与激活组比较显著降低(P<0.05);经LPS与100μmol.L-1VER共培养后,IL-17、IL-6和TGF-β1含量与未激活组和激活组比较均显著降低(P<0.05)。结论:外来病原体等刺激,使人淋巴细胞在激活状态下增殖、分化,引起机体免疫调节紊乱。Kv1.3通道阻断剂VER在体外可通过对人外周血淋巴细胞的直接作用,调节T淋巴细胞增殖、分化,使Th17/Treg细胞因子分泌发生改变。因此,推测T细胞膜Kv1.3钾离子通道可能为治疗自身免疫疾病的作用靶点之一。     

慢性阻塞性肺病稳定期患者外周血Th17/Treg失衡情况及与预后的关系

目的：观察慢性阻塞性肺病（chronic obstructive pulmonary disease,COPD）稳定期患者外周血中Th17与Treg细胞的水平.方法：选取本院2010年1月-2012年12月间确诊的40例COPD稳定期患者,选取同期40例健康体检健康人群作为对照组,收集两组人群的外周血后采用流式细胞术检测各个人群Th17与Treg细胞亚群的比例,采用ELISA法检测血清中的IL-17、TGF-β水平,采用RT-PCR检测转录因子RORγT与Foxp3 mRNA水平.结果：COPD稳定期患者外周血中Th17比例为（3.14±0.31）％,显著高于对照组人群的（1.23±0.24）％,差异有统计学意义（P＜0.05）;COPD稳定期患者外周血中Treg比例为（6.27±0.32）％,显著高于对照组人群的（3.43±0.32）％,差异有统计学意义（P＜0.01）;COPD稳定期患者外周血中IL-17水平为（89.36±4.13） pg/mL,显著高于对照组的（30.35±2.53） pg/mL,差异有统计学意义（P＜0.05）;COPD稳定期患者外周血中TGF-β水平为（681.45±41.65） pg/mL,显著高于对照组人群的（519.36±30.43） pg/mL,差异有统计学意义（P＜0.01）;COPD稳定期患者外周血转录因子Foxp3与RORγT mRNA水平均显著高于对照组人群,差异有统计学意义（均P＜0.01）.COPD稳定期患者Th17、Treg比例高水平的住院天数和病死率均高于对应低水平亚组（P＜0.05）.结论：Th17、Treg细胞及IL-17、TGF-β水平在COPD稳定期患者外周血表达增加,且转录因子Foxp3与RORγT mRNA水平表达增加,提示其与COPD患者的预后有关,其外周血中Th17与Treg细胞的不平衡可能参与了COPD发病和病程的进展.     

白花前胡丙素抗哮喘模型小鼠Th17/Treg失衡作用

目的 研究白花前胡丙素对哮喘模型小鼠体内Th17/Treg失衡的干预作用。方法 小鼠腹腔注射和雾化吸入卵清蛋白以建立气道炎性反应模型,并灌胃不同剂量的白花前胡丙素干预。模型结束后,动物肺功能仪分析气道高反应性;细胞计数器和Diff-Quick染色计数支气管灌洗液中白细胞总数及分类;酶联免疫吸附法检测血清或BALF中细胞因子和炎性介质的水平;苏木精-伊红(HE)染色法观察气道病理改变;流式细胞检测脾脏CD4+细胞数及分类;RT-PCR分析脾脏Foxp3 mRNA的表达。结果 与对照组比较,模型组呈现明显的气道炎性反应和气道高反应(P<0.05或P<0.01);IgE、IL-17水平增加(P<0.01),IL-10、TGF-β1水平降低(P<0.01);CD4⁺CD25⁺Foxp3⁺细胞占总CD4⁺细胞的比例降低(P<0.01);Foxp3 mRNA的表达减少(P<0.01)。与模型组比较,白花前胡丙素组气道炎症显著减轻,中、高剂量组气道反应在氯乙酰胆碱剂量>10 mg·kg^-1均显著受到抑制(P<0.05或P<0.01);白花前胡丙素组IgE水平显著降低(P<0.01),IL-10、TGF-β1水平显著增加(P<0.05或P<0.01),中、高剂量组IL-17显著降低(P<0.01);白花前胡丙素组显著增加CD4⁺CD25⁺Foxp3⁺细胞的比例(P<0.05或P<0.01),并增加Foxp3 mRNA的表达(P<0.05或P<0.01)。结论 白花前胡丙素具有抗哮喘鼠Th17/Treg失衡的作用。     

Systemic effects of acute cigarette smoke exposure in mice

Abstract

Context: Cigarette smoke (CS) causes both pulmonary and extrapulmonary disorders.

Objective: To determine the pulmonary and extrapulmonary effects of acute CS exposure in regard to inflammation, oxidative stress and DNA damage.

Materials and methods: Mice were exposed to CS for 10 days and then their lungs, heart, liver, pancreas, kidneys, gastrocnemius muscle and subcutaneous (inguinal and flank) and visceral (retroperitoneum and periuterus) adipose tissues were excised. Bronchoalveolar lavage fluid samples were obtained for differential cell analysis. Inflammatory cell infiltration of the tissues was assessed by immunohistochemistry for Mac-3⁺ cells, F4/80⁺ cells and CD45⁺ cells. Oxidative stress was determined by immunohistochemistry for thymidine glycol (a marker of DNA peroxidation) and 4-hydroxy hexenal (a marker of lipid peroxidation), by enzyme-linked immunosorbent assay for protein carbonyls (a marker of protein peroxidation) and by measurements of enzyme activities of glutathione peroxidase, superoxide dismutase and catalase. DNA double-strand breaks were assessed by immunohistochemistry for γH2AX.

Results: CS exposure-induced inflammatory cell infiltration, oxidative stress and DNA damage in the lung. Neither inflammatory cell infiltration nor DNA damage was observed in any extrapulmonary organs. However, oxidative stress was increased in the heart and inguinal adipose tissue.

Discussions: Induction of inflammatory cell infiltration and DNA damage by acute CS exposure was confined to the lung. However, an increased oxidative burden occurred in the heart and some adipose tissue, as well as in the lung.

Conclusions: Although extrapulmonary effects of CS are relatively modest compared with the pulmonary effects, some extrapulmonary organs are vulnerable to CS-induced oxidative stress.     

Ovarian toxicity of cigarette smoke exposure during pregnancy in mice

Pregnant mice of C57BL/6 and DBA/2 inbred strains were exposed to cigarette smoke during days 1-18 of pregnancy. Both mothers and offspring were killed at the age of 21-22 weeks, and ovaries were studied with respect to the number of primordial follicles and small primary follicles. In additional studies, 7,12-dimethylbenz(a)anthracene (DMBA) was administered on day 15 of pregnancy. Cigarette-smoke exposure during gestation did not affect the number of primordial follicles in the ovaries of mothers, whereas counts were significantly decreased (31%) in the ovaries of DBA/2 offspring, and decreased, although not significantly (20%) in the ovaries of C57BL/6 offspring. DMBA (5 to 50 mg/kg) decreased oocyte counts in the mothers in a dose-dependent manner. At the same doses, DMBA reduced the viability of the offspring. These studies suggest that primordial and small primary follicles may be more sensitive to components in cigarette smoke during fetal life than in adulthood.     

桔梗总皂苷调控Th17/Treg免疫失衡改善哮喘模型小鼠气道炎症

目的 探究桔梗总皂苷是否通过介导Notch通路对哮喘Th17/Treg平衡进行调控。方法 48只BALB/c小鼠随机分为正常对照组,模型组,桔梗总皂苷低、中、高剂量组(15,30,60 mg·kg^-1)和地塞米松组,每组8只。雾化吸入卵清蛋白建立小鼠哮喘模型,药物干预8周后,测定小鼠气道高反应性。取材支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)细胞计数、ELISA检测、HE、PAS及Masson染色,流式细胞术检测小鼠肺组织Th17、Treg细胞百分比及Th17/Treg比值变化情况,qPCR检测小鼠肺组织Notch1、Jagged1、RORγt、Foxp3表达水平,Western blotting检测小鼠肺组织中Notch1、Jagged1、Hes1及RORγt、Foxp3、IL-10、IL-17蛋白表达情况。结果 与模型组相比,桔梗总皂苷中、高剂量组小鼠气道阻力减小,BALF中炎性细胞数量及炎性因子IL-4、IL-5、IL-13含量降低,血清IL-17、IL-6、IgE含量降低,BALF中INF-γ及血清中IL-10含量水平显著升高(P<0.05或P<0.01)。HE、PAS及Masson结果显示,桔梗总皂苷高剂量组及地塞米松组能够显著缓解炎性细胞浸润及支气管膜破坏情况,减少杯状细胞增生,同时减轻组织胶原纤维散出程度。流式细胞术结果显示,桔梗总皂苷中、高剂量组Th17细胞数量及Th17/Treg占比均显著减少,Treg细胞数量显著增加(P<0.05或P<0.01)。qPCR及Western blotting结果显示,桔梗总皂苷给药处理后小鼠肺组织Notch1、Jagged1、RORγt mRNA及Notch1、Jagged1、Hes1、RORγt、IL-10、IL-17蛋白的表达水平均显著降低,Foxp3 mRNA及Foxp3蛋白的表达水平显著升高(P<0.05或P<0.01)。结论 桔梗总皂苷能够介导Notch通路对哮喘Th17/Treg平衡进行调控,从而起到减轻气道炎症,抑制哮喘发作的作用。     

青蒿素对变应性接触性皮炎小鼠Treg/Th17免疫平衡的影响

目的探讨青蒿素(Art)在变应性接触性皮炎(ACD)中对Th17和Treg的影响及机制。方法以DNFB致敏和激发制作小鼠ACD模型,局部外涂Art进行干预,采用RT-PCR和ELISA检测Treg/Th17细胞特异性核因子及相关细胞因子的表达与含量,流式细胞术检测Treg细胞数量,West-ern blot方法检测信号转导子和转录激活子STAT3的磷酸化活性表达。结果与正常组相比,模型组IL-6、IL-17含量、ROR-γt及STAT3的活性表达明显升高,同时Foxp3表达降低。Art明显降低ROR-γt表达、下调IL-6及IL-17水平,同时增加Foxp3表达并促进Treg产生,减弱STAT3的磷酸化活性。结论 Art可能通过调节Treg/Th17免疫平衡,对DN-FB诱导的ACD小鼠发挥免疫治疗作用。     

Th17/Treg平衡在类风湿关节炎中作用的研究进展

类风湿性关节炎(rheumatoid arthritis,RA)是一种以慢性多关节滑膜炎为主要特征的自身免疫性疾病。虽然目前对于RA的确切发病机制尚不明确,但一般认为和T细胞相关。最近研究发现调节性T细胞(regulatory T cell,Treg)和Th17细胞在RA的发生发展中发挥重要作用。Th17细胞能够分泌促炎症因子IL-17,通过诱导基质金属蛋白酶(ma-trix metallo proteinases,MMPs)和破骨细胞生成,促进骨滑膜炎症、骨和关节损伤;而Treg则通过释放抑制性细胞因子IL-10和TGF-β发挥免疫效应,调控RA中的炎症性免疫应答过程。单独TGF-β作用下诱导初始T细胞分化为Treg,而在TGF-β和IL-6共同作用下诱导初始T细胞分化为Th17细胞,因此,Th17和Treg细胞在特定的细胞因子微环境下可以相互转化。调节Th17/Treg之间的平衡可能成为治疗RA的新方法。该文将对Th17/Treg平衡在RA发生发展中的调节作用作一综述。     

芍药甘草汤对哮喘大鼠Treg/Th17失衡及其相关细胞因子的干预作用

目的 观察芍药甘草汤对支气管哮喘SD大鼠Treg/Th17失衡及相关细胞因子IL-2、IL-6、IL-10、IL-17的干预情况,探讨芍药甘草汤防治哮喘的作用机制。方法 SD大鼠40只,♂,随机均分为正常对照组、模型组、芍药甘草汤组、地塞米松组、中西医结合组,每组8只。除正常对照组外其余各组大鼠以卵白蛋白致敏并诱发哮喘模型。正常对照组和模型组生理盐水灌胃,其余组给予对应药物,用Elisa酶联免疫法测定肺匀浆中相关细胞因子IL-2、IL-6、IL-10、IL-17水平。流式细胞仪检测大鼠脾脏CD4⁺细胞数及比例。结果 与模型组比较,各药物组均可显著降低IL-2、IL-6及IL-17水平(P<0.01),同时能显著升高IL-10含量(P<0.01);各药物组之间比较IL-2、IL-6和IL-10差异无统计学意义(P>0.05);地塞米松组IL-17表达比芍药甘草汤组及中西医结合组均显著降低(P<0.05)。与模型组比较,芍药甘草汤组能显著增加CD4⁺CD25⁺Foxp3⁺Treg细胞的比例(P<0.01)。结论 芍药甘草汤防治哮喘的作用机制可能与调节哮喘大鼠Treg/Th17比例及其相关细胞因子表达的作用有关。     

Periplocoside A ameliorated type II collagen-induced arthritis in mice via regulation of the balance of Th17/Treg cells

Periplocoside A (PSA) has been extracted from the Chinese herbal medicine Periploca sepium Bge to treat rheumatoid arthritis (RA) via immune regulation. We previously found that PSA exhibits immunosuppressive activity both in vitro and in vivo. Balanced regulation of helper T 17 (Th17)/regulatory T (Treg) cells is the current therapeutic direction for the treatment of RA. The present study investigated the mechanism of PSA in treating collagen-induced arthritis (CIA). The therapeutic effects and potential pharmacological mechanisms of PSA were specifically clarified by examining its effects on CIA in DBA/1 mice. PSA administration significantly relieved the severity of the arthritis, and preventive administration of PSA reduced the incidence of arthritis in the mice with CIA and relieved joint damage in terms of morphology. PSA was also able to reduce the levels of anti-collagen II (CII) antibodies and pro-inflammatory cytokines in the serum. As a result, the proportion of Th17 cells decreased, and the proportion of Treg cells increased. A follow-up study of the ex vivo immunological reactions induced by a specific antigen found that PSA suppressed lymphocyte proliferation, inhibited the differentiation and reactivity of Th17 cells, and promoted the proportion of Treg cells among helper T cells. PSA also exhibited pharmacological effect in regulating the balance between Th17 and Treg cells in CIA through relevant signalling pathways. Thus, PSA played a specific role in CIA treatment. In particular, our results suggest that the therapeutic effects of PSA on RA are partially realized via the regulation of the balance of Th17/Treg cells.     

外周血辅助性T细胞17调节性T细胞及二者比值在系统性红斑狼疮的研究

目的探讨Th17细胞和调节性T细胞在系统性红斑狼疮(SLE)、狼疮肾炎(LN)发生和发展中的作用,以期寻找病情评估的无创性新指标。方法选取SLE患者103例及23名健康体检者,其中LN患者40例,新诊断、未治疗的SLE患者42例。SLE患者非活动组(SLEDAI 0~4分)患者16例、轻度活动组(SLEDAI 5~9分)患者29例、中度活动组(SLEDAI 10~14分)患者26例、重度活动组(SLEDAI≥15分)患者32例。用四色分选流式细胞仪检测外周血CD4+IL-17+Th17细胞和CD4+CD25+FOXP3+Treg细胞百分数,分析其与疾病活动性(SLEDAI评分)、补体C3、C4和肾脏受累的关系。结果 1SLE轻度、中度、重度活动组Th17细胞百分数均高于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05);SLE中度、重度活动组Th17细胞均高于SLE轻度活动组,差异均有统计学意义(P<0.05);LN组Th17细胞高于非LN组,差异有统计学意义(P<0.05);新发SLE活动组Th17细胞百分数高于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05)。补体降低组SLE患者Th17细胞高于补体正常组及健康对照组,差异均有统计学意义(P<0.05)。Th17细胞百分数与SLEDAI评分呈正相关。2SLE中度、重度活动组Treg细胞百分数均低于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05);SLE重度活动组Treg细胞低于SLE轻度活动组,差异有统计学意义(P<0.05)。LN组Treg细胞与非LN组差异无统计学意义。新发SLE活动组Treg细胞百分数低于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05)。补体降低组SLE患者Treg细胞低于补体正常组及健康对照组,差异均有统计学意义(P<0.05)。Treg细胞百分数与SLEDAI评分无相关性。3SLE轻度、中度、重度活动组Th17/Treg比值均高于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05);SLE中度、重度活动组Th17/Treg比值均高于SLE轻度活动组,差异均有统计学意义(P<0.05);LN组Th17/Treg高于非LN组,差异有统计学意义(P<0.05)。新发SLE活动组Th17/Treg细胞比值高于健康对照组和SLE非活动组,差异均有统计学意义(P<0.05)。补体降低组SLE患者Th17/Treg比值高于补体正常组及健康对照组,差异均有统计学意义(P<0.05)。Th17/Treg比值与SLEDAI评分呈正相关。结论活动性SLE患者外周血Th17细胞升高、Treg细胞下降,Th17/Treg细胞比值与疾病活动呈正相关,提示SLE发病不仅仅是Th17细胞升高或Treg细胞下降,而是二者相互作用导致免疫失衡导致。     

Pharmacodynamic effects of chronic cigarette smoke exposure in spontaneously hypertensive rats

In investigating the influence of chronic cigarette smoke exposure on hypertension, we compared the pharmacodynamic effects of enforced exposure to smoke on spontaneously hypertensive rats (SHR) with those on Wistar-Kyoto (WKY) rats. Chronic cigarette smoke exposure for 8 weeks decreased the elevated heart rate of mature male SHR to approximately the rate in WKY rats 24 h after smoke exposure. Both systolic and diastolic blood pressures also decreased slightly. However, WKY rats showed a marked rise in heart rate soon after exposure to cigarette smoke began, with no change in blood pressure, while the heart rate of SHR in the early stage remained similar to that of animals without exposure, although their blood pressure was clearly reduced. The body weight of both strains tended to decrease during smoke exposure, but the effect was more severe in SHR. Moreover, the effects of chronic smoke exposure were observed using retired, aged female SHR breeders. A decrease in body weight and heart rate, but not in blood pressure, was also recognized even in these mature animals. These effects gradually recovered after withdrawal from exposure. On the basis of these results, a profile of chronic cigarette smoke exposure under hypertension is discussed in this study.     

他克莫司对白癜风小鼠Th17/Treg细胞平衡及黑素细胞丢失的影响

目的 探究他克莫司对白癜风小鼠辅助性T17（Th17）/调节性T （Treg）细胞平衡和黑素细胞丢失的影响。方法 将小鼠随机分为对照组、模型组和他克莫司低、中、高剂量组（涂抹10、50、100 mg 0.03%他克莫司软膏）,使用松香/蜡混合物对小鼠背部相同位置进行去毛,大小为2 cm×2 cm,将其分为两部分：用药区域与非用药区域,面积1：1。除对照组外,于小鼠背部用药区均匀涂抹50 mg 40%莫诺苯宗软膏,每天1次,连续90 d,制备白癜风模型。莫诺苯宗涂抹30 d后开始于相同位置涂抹他克莫司软膏,2种药膏涂抹时间间隔7 h,他克莫司软膏每天早晚各涂抹1次,连续用药60 d。肉眼观察小鼠皮毛脱色情况并进行脱色评分,通过反射式共聚焦显微镜（RCM）观察小鼠皮肤黑素细胞和黑色素的分布,取小鼠皮损进行苏木精-伊红染色（HE）和马松染色（MF）对基底层黑素细胞和含黑色素的毛囊进行计数,通过流式细胞术测定小鼠外周血中Th17和Treg淋巴细胞的比例,酶联免疫吸附试验（ELISA）检测外周血白细胞介素-17（IL-17）、IL-22和叉头型基因p3（Foxp3）的含量。结果 与对照组比较,白癜风模型组小鼠用药区皮毛明显脱色,脱色评分显著升高（P<0.05）,皮损处色素明显缺失;含黑色素的毛囊和黑素细胞显著减少（P<0.05）;Th17/Treg淋巴细胞比例显著升高（P<0.05）;血清IL-17、IL-22水平显著上升,Foxp3水平显著降低（P<0.05）。与模型组比较,他克莫司各组小鼠皮毛脱色情况明显改善,脱色评分显著降低（P<0.05）,皮损处可见色素分布及黑素细胞;含黑色素的毛囊和黑素细胞显著增多（P<0.05）;Th17/Treg淋巴细胞比例显著降低（P<0.05）;IL-17、IL-22水平显著降低,Foxp3水平显著升高（均P<0.05）。结论 他克莫司可调控白癜风小鼠Th17/Treg淋巴细胞平衡,抑制其黑素细胞丢失。     

3-week inhalation exposure to cigarette smoke and/or lipopolysaccharide in AKR/J mice

AKR/J mice were exposed to cigarette smoke (CS) and/or lipopolysaccharide (LPS) via inhalation for 3 wk and pulmonary responses were evaluated. The objective was to explore the feasibility of coexposing LPS with cigarette smoke under a subacute exposure, as a surrogate for viral or bacterial insults, that would mimic the pathogenesis of infection-related chronic obstructive pulmonary disease (COPD) exacerbations. The study was the first step in an effort to develop a rodent COPD model in which morphologic lesions of COPD develop in a shorter period of exposure and more closely simulate human COPD. Mice were exposed 6 h/day, 5 days/wk for 3 wk to one of the following: (1) sham control: filtered air; (2) CS: 250 microg/L wet total particulate matter (WTPM) for 5 h/day followed by 1 h/day air; (3) LPS: 0.5 microg/L LPS (055:B5 Escherichia coli; 3,000,000 EU/mg) for the last 1 h/day 2 day/wk (following 5 h/day of filtered air); and (4) CS/LPS: CS 5 h/day followed by air or LPS (2 days/wk) for 1 h/day. After the last exposure, animals were necropsied and subjected to bronchoalveolar lavage (BAL) or histopathology. The BAL neutrophil counts were highest in the LPS group, while macrophage counts were higher in the CS/LPS group than other exposed groups. The LPS group displayed the greatest increases in BAL cytokines, while KC (keratinocyte-derived chemokine) and TARC (thymus and activation-regulated chemokine) were highest in the CS group. The CS/LPS group had generally lower cytokine levels relative to the LPS or CS groups, except for the levels of RANTES and G-CSF (granulocyte-colony stimulating factor) comparable to the LPS group. At microscopic examination of lung sections, cellular inflammatory infiltrates were most notable in the CS/LPS group, which had a diffuse, predominantly macrophage infiltrate with fewer neutrophils. The LPS group had predominantly neutrophils in the pulmonary infiltrate and the CS group had a predominantly macrophage infiltrate in alveolar ducts and adjacent alveoli. Apoptotic labeling of lung cells was highest with the CS/LPS group. In summary, the CS/LPS group displayed greater cellular infiltration and apoptotic responses in the lung with an indication of immunosuppressive effects (lower BAL cytokines) than the CS or LPS group, suggesting that the CS/LPS model shows promise to be further explored as an animal model for studying pathogenesis of COPD exacerbations. A longer term study with interim assessments is needed to confirm that the subacute responses observed in the CS/LPS group will result in greater severity of COPD-related pulmonary lesions following prolonged exposures.     

白藜芦醇调控Th17/Treg抑制血吸虫病肝脏肉芽肿

目的探讨白藜芦醇对血吸虫病肝脏肉芽肿作用,分析其对Th17、Treg应答的影响。方法 45只C57BL/6小鼠感染日本血吸虫3周后,随机分为感染组、灌胃白藜芦醇治疗组、灌胃吡喹酮治疗组,另取15只正常小鼠为健康对照组。感染第9周,取肝脏HE染色观察虫卵肉芽肿,计数中性粒细胞;RT-PCR检测肝脏中IL-17A、Foxp3、CXCL1、CXCL2水平;流式细胞术检测脾脏中Th17、Treg应答。用PBS、SEA刺激正常小鼠脾脏细胞,分别加入白藜芦醇和吡喹酮,流式细胞术检测Th17、Treg应答。结果与感染组相比,白藜芦醇治疗组,肝脏肉芽肿减轻,肉芽肿中性粒细胞数量减少(P<0.01),Th17应答减弱(P<0.05),Treg应答增强(P<0.05),肝脏IL-17A、CXCL1、CXCL2表达均降低(P<0.05),Foxp3表达增高(P<0.01)。白藜芦醇促使SEA刺激的T细胞往Th17细胞分化减少(P<0.05),往Treg细胞分化增多(P<0.05)。结论白藜芦醇通过降低Th17应答,增强Treg应答,抑制血吸虫病肝脏肉芽肿。     

Stress and hypothermia in mice in a nose-only cigarette smoke exposure system

In nose-only exposure systems, animals need to be restrained inside a tube, which leads to stress. Stress is known to cause hyperthermia in rodents. Chronically repeated episodes of hyperthermia could be detrimental to animal health and influence results of nose-only exposure studies. Therefore we investigated whether hyperthermia occurred in male C57BL/6J mice that were restrained for increasing lengths of time, using nosepieces held at room temperature, preheated at 37 degrees C, or thermostat controlled at different temperatures, with and without exposure to different concentrations of cigarette smoke. Body temperature, body weight, plasma corticosterone levels, and adrenal weights were recorded. Restraint using nosepieces at room temperature caused a time-dependent decrease in body temperature, which could be reversed by preheating the nosepieces to 37 degrees C. Cigarette smoke dose-dependently caused an additional decrease, which was counteracted by controlling nosepiece temperature at 38 degrees C. During 3 mo of exposure using heated nosepieces, Delta body temperature remained constant. Body weight gain did not differ between smoke-exposed and room air-breathing animals exposed using either heated or room-temperature nosepieces, but both groups gained significantly less weight, while adrenal weights were significantly and similarly increased, when compared to unrestrained littermates. Plasma corticosterone levels did not differ between the three groups. In conclusion, during restraint in nose-only exposure tubes with room temperature metal nosepieces, mice suffer a pronounced hypothermia. Preventing this by heating the nosepieces does not reduce the stress experienced by the animals.     

慢性丙肝及肝硬化患者外周血Th17/Treg细胞及其相关细胞因子的表达及意义

目的研究慢性丙肝及肝硬化患者外周血Th17细胞、Treg细胞及其相关因子的表达及意义,分析Th17细胞、Treg细胞及其相关因子的变化对患者的影响。方法选取2014年3月~2016年2月在运城市中心医院进行治疗的慢性丙肝患者47例,肝硬化患者27例,同时从健康人群中选取39人,分成慢性丙肝组、肝硬化组和健康对照组。用流式细胞检测三组患者Th17细胞和Treg细胞表达率,用ELSIA技术检测IL-10、IL-17、IL-6、TGF-β的血清水平,比较三组之间的差异。结果健康对照组Th17细胞表达率及IL-6、IL-17水平明显低于慢性丙肝组,健康对照组Treg细胞表达率及IL-6水平明显低于慢性丙肝组和肝硬化组,健康对照组和慢性丙肝组Th17/Treg比值明显高于肝硬化组,健康对照组和慢性丙肝组TGF-β、IL-10水平明显低于肝硬化组,差异均有统计学意义(P<0.05)。结论 Th17细胞和Treg细胞表达率以及相关因子的血清水平的变化与慢性丙肝及肝硬化有密切的关系,关注Th17细胞和Treg细胞表达率以及相关因子的血清水平的变化对预防慢性丙肝及肝硬化有重要指导意义。     

Notch信号通路调控Treg/Th17细胞机制的研究进展

摘要： Notch家族是多细胞生物发育过程中一类高度保守的、 十分重要的跨膜信号蛋白, 通过与相邻细胞之间的相互作用, 在细胞增殖、 分化、 凋亡中发挥着关键作用。调节性T细胞 （Treg） 及辅助性T细胞17 （Th17） 是目前发现的一类新型CD4+ T细胞亚群, 在生理状态下, 两者可通过分泌多种细胞因子来调节机体免疫的平衡。近年来, 越来越多的研究发现Notch信号通路通过调控Treg、 Th17细胞参与机体多种疾病的发生。本文就Notch信号通路在血液系统疾病、 自身免疫系统疾病等疾病中对Treg/Th17细胞调控机制的研究进展作一简要综述。     

In vitro exposure to cigarette smoke induces oxidative stress in follicular cells of F1 hybrid mice

This study assessed the influence of cigarette smoke condensate (CSC) and benzo(a)pyrene [B(a)P] on the levels of two oxidative stress biomarkers [8‐isoprostane (8‐IsoP) and 8‐hydroxy‐2‐deoxy Guanosine (8‐OH‐dG)], in in‐vitro spent media of follicle cells. Follicles (100–130 µm) isolated from ovaries of F₁ hybrid (C57Bl/6j × CBA/Ca) mice were cultured for 13 days in media exposed to B(a)P [0 ng ml^–1 (control) to 45 ng ml^–1] or CSC [0 µg ml^–1 (control) to 130 µg ml^–1]. The concentrations of oxidative stress biomarkers in spent media were quantified by enzyme‐linked immune sorbent assays (ELISA). CSC and B(a)P treatment induced a significant, dose‐dependent increase in the concentrations of 8‐IsoP and 8‐OH‐dG in the spent media. We conclude that CSC and B(a)P exposure can induce oxidative stress in ovarian follicles, an effect that may contribute to the previously documented decline in follicle development and premature ovarian failure in women who smoke. © Her Majesty the Queen in Right of Canada 2013.