参麦注射液对支气管哮喘患者外周血Th17/Treg细胞的影响

目的 探讨参麦注射液对支气管哮喘(简称哮喘)患者外周血Th17/Treg细胞的影响及其意义.方法 选取山西医科大学第一医院呼吸科急性发作期、缓解期哮喘患者各25例,选取山西医科大学第一医院体检中心肺功能正常、激发或者舒张试验阴性的健康人25例作为对照组,分别使用1640培养液及参麦注射液进行干预处理.抽取晨起空腹静脉血5 ml,运用流式细胞仪对各组外周血Th17细胞、Treg细胞分别占CD4+的比例及Th17/Trcg细胞的比值进行比较,同时检测血清中IL-10因子的水平.结果 ①各组间干预前CD4+ Th17细胞/CD4+T细胞、CD4+ Th17细胞/CD4+ Treg细胞的比值,急性发作期组(3.05±1.27、1.49±1.78)高于缓解期组(2.38±0.93、0.61±0.24)及对照组(1.19±0.39、0.30±1.14),差异有统计学意义(P＜0.05),缓解期高于对照组,差异有统计学意义(P＜0.05);CD4+Treg细胞/CD4+T细胞的比值,对照组(5.20±3.26)高于缓解期组(3.99±0.90)及急性发作期组(2.76±0.93),差异有统计学意义,(P＜0.05),缓解期组高于急性发作期组,差异有统计学意义(P ＜0.05).血清中IL-10因子的水平,对照组(5,28±1.31)高于缓解期组(4.46±0.83)及急性发作期组(3.90±0.64),差异有统计学意义(P＜0.05),缓解期组高于急性发作期组,差异有统计学意义(P ＜0.05).②同一组间CD4+ Th17细胞/CD4+T细胞、CD4+Th17细胞/CD4+ Treg细胞的比值,干预前高于干预后,差异有统计学意义(P＜0.05).CD4+ Treg细胞/CD4+T细胞的比值,干预后高于干预前,差异有统计学意义(P＜0.05).血清中IL-10因子水平,干预后高于干预前,差异有统计学意义(P ＜0.05).结论 哮喘患者外周血中存在Th17/Treg细胞失衡,参麦注射液可以改善哮喘患者的Th17/Treg细胞失衡.     

Th1/Th2漂移与支气管哮喘的免疫治疗

Th1/Th2细胞在不同的细胞因子、抗原等因素的影响下，可发生Th1/Th2的转换，研究表明在支气管哮喘（哮喘）的发生机制中，Th2细胞分化明显增多，而Th1细胞数目减少。因此，促进Th1反应，抑制Th2反应的免疫治疗是治疗哮喘的重要部分。目前许多调节Th1/Th2失衡的免疫治疗方法已取得一些进展。     

支气管哮喘的免疫发病机制——从Th1/Th2细胞失衡到Th2/Treg细胞失衡

支气管哮喘(简称哮喘)是最常见的呼吸道慢性疾病之一,近年来随着全球哮喘防治创议(GINA)指南的全面推广,哮喘的防治取得了很大的进展。但迄今为止哮喘仍然不能得到根治,还存在对包括吸入肾上腺糖皮质激素在内的综合治疗     

布地奈德对支气管哮喘患者Th17/Treg平衡的干预机制研究

目的探讨布地奈德对支气管哮喘(简称哮喘)患者Th17/TregT细胞平衡的干预机制。方法收集轻、中、重度哮喘患者各15例用布地奈德进行干预治疗,健康者15人作为对照组,观察治疗前后外周血Th17及其胞内细胞因子IL-17和IL-23,Treg及其胞内细胞因子IL-10和TGF-β1;同时对以上观察对象进行肺功能检测。结果①Th17/Treg:中及重度哮喘患者Th17及其IL-17、IL-23显著高于健康对照组,而Treg及其IL-10、TGF-β1显著低于健康对照组;经布地奈德干预后Th17及其细胞因子下降,而Treg及其细胞因子回升。②肺功能:治疗后轻及中度患者的FEV1显著回升,且FEV1下降及回升率以及PEF波动率下降,而重度组治疗后上述效果不明显。③相关性分析:Th17与Treg呈显著负相关性。Th17与FEV1呈负相关性;与FEV1下降及上升率、呼吸峰流速波动率呈正相关性。而Treg与FEV1呈正相关性;与FEV1下降及上升率、呼吸峰流速波动率呈负相关性(P<0.05)。结论布地奈德可有效抑制哮喘患者Th17的数量与功能,同时增强Treg平衡数量与功能,使平衡向Treg回落,并且上述改变与哮喘的病变程度及肺功能的恢复紧密联系。     

大鼠支气管哮喘模型γδT细胞Th1/Th2免疫应答模式的研究

目的　探讨γδT细胞在哮喘的免疫应答模式 ,认识γδT细胞亚群在哮喘发病机制中的作用。方法　Wistar大鼠 2 0只 ,随机分为健康对照组与哮喘组 (用鸡卵清蛋白致敏和刺激大鼠 ,制作哮喘模型 ) ,每组 10只。收集外周血单个核细胞 (PBMC)和支气管肺泡灌洗液 (BALF) ,用补体攻击法结合洗淘法选择性培养扩增γδT细胞 ,并用流式细胞术鉴定培养体系中的γδT细胞及其纯度 ,用原位杂交法测γδT细胞白细胞介素 (IL) 4mRNA和干扰素 (IFN)γmRNA的表达 ,用ELISA检测培养上清液中IL 4和IFN γ的浓度。结果　哮喘组大鼠PBMC和BALF中 ,γδT细胞培养上清液中IL 4浓度显著高于健康对照组 (P <0 0 1) ,IFN γ浓度低于健康对照组 (P <0 0 1) ;γδT细胞IL 4mRNA表达阳性率高于健康对照组 (P <0 0 1) ,IFN γmRNA表达阳性率低于健康对照组 (P <0 0 1)。结论γδT细胞或者γδT细胞亚群存在Th1/Th2模式 ,在大鼠哮喘模型呈Th2优势应答 ,γδT细胞参与了哮喘的发病过程。     

大鼠胰岛B细胞功能对Th1/Th2淋巴细胞分化的影响

目的　探讨大鼠胰岛B细胞功能对Th1/Th2淋巴细胞分化的影响及其机制。方法　 2 0 0 2 - 11～ 2 0 0 3- 0 9在北京大学人民医院将 4 0只雄性SD大鼠随机分A、B、C、D和E组 ,每组 8只。A组 :腹腔注射链脲菌素(STZ)建成 1型糖尿病模型 ;B组 :腹腔注射卵清蛋白 (OVA )致敏 OVA激发建成哮喘大鼠模型 ;C组 :腹腔注射STZ OVA致敏和激发 ;D组 :腹腔注射STZ 皮下注射胰岛素 OVA致敏和激发 ;E组为对照组。检测各组血清C肽浓度、支气管肺泡灌洗液 (BALF)IL 4和IFN γ质量浓度。结果　与对照组比较 ,A、C组的血清C肽浓度显著下降 ,而BALF的IFN γ质量浓度显著增高 ,IL 4质量浓度显著下降 ;与C组比较 ,D组经注射胰岛素后 ,其BALF中的IFN γ质量浓度显著降低 (137 6 3± 9 94vs 111 0 0± 12 14 ,P <0 0 5 ) ,IL 4质量浓度显著增高 (11 0 0± 3 93vs15 6 3± 2 6 7,P <0 0 5 )。结论　大鼠胰岛B细胞功能对Th1/Th2淋巴细胞的分化有调节作用。     

茶碱对支气管哮喘患者血中T淋巴细胞亚群的影响

支气管哮喘的发病机制较为复杂 ,治疗哮喘药物的作用机理亦未完全明确 ,茶碱是治疗支气管哮喘的首选药物之一 ,对哮喘有良好的治疗作用。本组资料对支气管哮喘急性发作患者茶碱治疗前后 T淋巴细胞亚群的变化进行观察。对象与方法1.对象 :( 1)支气管哮喘急性发作轻度患者 3 0例 ,均符合支气管哮喘防治指南诊断标准[1 ] ,男性 12例 ,女性 18例 ,年龄19～ 5 3岁 ,近期未用肾上腺皮质激素及茶碱类药物。上述 3 0例支气管哮喘患者给予茶碱治疗 3周 (初期静滴加口服 ,逐渐改为单纯口服 ) ,病情符合支气管哮喘防治指南非急性发作期轻度哮喘论断标…     

喘可治注射液逆转哮喘患者Th1/Th2失衡的作用机制

采集发作期哮喘患者的静脉血,分离外周血单个核细胞（PBMC）,分为两份,一份加入PHA（对照组）,另一份加入喘可治注射液和PHA（观察组）,培养48h后分别收集上清液和细胞沉淀,采用RT-PCR方法测定细胞沉淀中T-bet mRNA、GATA-3 mRNA的表达强度,ELISA法测定上清液中IL-4、IL-5、IL-13、IFN-γ的表达水平。结果：①观察组细胞沉淀中T-bet mRNA的表达强度较对照组显著增强,GATA-3 mRNA表达强度显著减低;T-bet／GATA-3比值较对照组显著增高。②观察组上清液中IL-4、IL-5、IL-13水平均较对照组显著降低,IFN-γ水平较对照组显著增高。认为喘可治注射液可通过上调T-bet表达、下调GATA-3表达来调节T-bet／GATA-3比值,逆转Th1／Th2细胞失衡。     

布地奈德雾化吸入对轻中度支气管哮喘患者Th1/Th2细胞因子平衡的影响

目的探讨雾化吸入布地奈德对轻中度支气管哮喘患者Th1/Th2细胞因子平衡的影响。方法49例轻中度支气管哮喘患者随机分为对照组(25例)和治疗组(24例)。两组均予吸氧、抗感染、静脉氨茶碱及对症治疗等。治疗组加用布地奈德混悬液氧气雾化吸入。分别采用ELISA法检测两组哮喘患者治疗前、治疗两周后及健康体检者外周血浆白细胞介素-5(inter-leuk in-5)和IL-12水平;同时测定两组哮喘患者治疗前后的FEV1。结果两组哮喘患者治疗前后血浆IL-5水平明显高于健康体检组,而IL-12水平则明显低于健康体检组,均P0.05;治疗后两哮喘组IL-5、IL-12水平与治疗前比较,均P0.01,治疗组与对照组治疗后比较IL-5、IL-12亦均差异显著;无论是治疗前还是治疗后,两组哮喘患者血浆IL-5与IL-12均呈明显的直线副相关。结论Th1/Th2的失衡与哮喘发病密切相关;布地奈德雾化吸入可通过调节Th1/Th2的平衡而治疗哮喘。     

支气管哮喘患者急性发作期外周血LncRNA NEAT1表达及与Th1/Th2平衡的关系

目的：探讨支气管哮喘（BA）患者急性发作期外周血长链非编码RNA（LncRNA）核富集转录本1（NEAT1）表达水平及其与辅助性T细胞1（Th1）/辅助性T细胞2（Th2）平衡的关系。方法：选取BA缓解期患者36例（缓解期组）、急性发作期患者44例（急性发作期组）,另选取同期体检健康者50例作为对照组。利用实时荧光定量PCR（qRT-PCR）测定外周血LncRNA NEAT1表达,应用流式细胞仪测定外周血单个核细胞（PBMC）中Th1、Th2细胞比例,应用ELISA法测定外周血干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）水平;采用Pearson检验分析BA急性发作期患者外周血LncRNA NEAT1表达与Th1/Th2、IFN-γ/IL-4比值的关系,并绘制ROC曲线分析LncRNA NEAT1表达水平对BA患者急性发作的预测价值。结果：急性发作期组患者外周血LncRNA NEAT1表达水平最高,其次为缓解期组,对照组最低,组间比较差异有统计学意义（均P＜0.05）。PBMC中Th1细胞比例及Th1/Th2比值高低依次为对照组、缓解期组、急性发作期组,Th2细胞比例高低依次为急性发作期组、缓解期组、对照组,组间比较差异有统计学意义（均P＜0.05）。外周血IFN-γ水平及IFN-γ/IL-4比值高低依次为对照组、缓解期组、急性发作期组,IL-4水平高低依次为急性发作期组、缓解期组、对照组,组间比较差异有统计学意义（均P＜0.05）。BA急性发作期患者外周血LncRNA NEAT1表达与Th1/Th2、IFN-γ/IL-4比值呈负相关（均P＜0.05）。外周血LncRNA NEAT1表达水平预测BA患者急性发作的曲线下面积为0.903,最佳截断值为1.73,敏感性为81.80%,特异性为88.90%。结论：BA患者急性发作期外周血LncRNA NEAT1高表达,可能与Th1/Th2平衡紊乱有关。     

乌司他丁联合参麦注射液治疗ALI的疗效研究

目的观察乌司他丁联合参麦注射液对急性肺损伤的疗效。方法将60例患者随机分为治疗组和对照组,每组30例,两组常规治疗相同。治疗组加乌司他丁、参麦注射液静滴。观察两组患者临床疗效、治疗前后呼吸频率(RR)及血气分析指标,并测定血浆TNF-α及SOD的含量。结果治疗组临床疗效优于对照组;治疗3天后,治疗组患者RR、PaO2、PaO2/FiO2均优于对照组;治疗3天后始,随着治疗时间的延长,两组TNF-α及SOD检测值均呈下降趋势,治疗组的TNF-α较对照组下降更明显,而治疗组SOD的活力明显高于对照组。结论乌司他丁联合参麦注射液治疗ALI患者,有利于抑制炎症因子、氧自由基生成,临床疗效显著。     

非小细胞肺癌患者Th1/Th2型细胞因子的表达及其临床意义

目的研究辅助性T淋巴细胞(helper T lymphocyte,Th)1、2型细胞因子在非小细胞肺癌(non-small cell lung canc-er,NSCLC)患者外周血中的表达。方法采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测40例NSCLC患者手术前后和20例正常对照外周血中Th1型细胞因子白细胞介素-2(interleukin-2,IL-2)、肿瘤坏死因子-α(tumor necrosis fac-tor-α,TNF-α)和Th2型细胞因子IL-10、转化生长因子-β1(transforming growth factor-β1,TGF-β1)的水平。结果 NSCLC组术前IL-2、TNF-α水平较正常对照组明显降低(P<0.05);IL-10、TGF-β1水平则明显升高(P<0.05)。NSCLC患者手术后IL-2、TNF-α水平较手术前明显升高(P<0.01);IL-10、TGF-β1水平则明显降低(P<0.01)。外周血Th1/Th2型细胞因子水平与NSCLC临床病理无关。结论①NSCLC患者呈现Th1抑制和Th2极化状态,是肺癌发生免疫耐受的可能机制;②手术切除肿瘤可在一定程度上逆转Th1/Th2的失衡状态,改善机体免疫状态,对预后产生积极作用。     

Peripheral Blood Th1 and Th2 Profile in Patients with Moderate Asthma: Effect of Inhaled Corticosteroid

The peripheral blood from healthy subjects and asthma patients was stimulated with phorbol 12-myristate 13-acetate and ionomycin, and the cells were stained with anti-CD4 antibody, permeabilized, stained with anti-IFN-γ and anti-IL-4 antibodies, and analyzed by flow cytometry. Compared with healthy subjects, asthma patients showed a greater percentage of both IL-4(+) IFN-γ (–) CD4 cells (Th2 cells) and IFN-γ(+) IL-4(–) CD4 cells (Th1 cells). The percentage of Th2 cells was correlated with serum IgE level. After treatment with inhaled corticosteroid, Th2 cells decreased at week 24, but not week 4. Long-term therapy with inhaled steroid may thus be required for improvement in lymphocytic inflammation.     

Suplatast Tosilate Alters DC1/DC2 Balance in Peripheral Blood in Bronchial Asthma

Suplatast tosilate is an antiallergic drug that selectively suppresses Th2-cytokine production and inhibits airway hyperresponsiveness and eosinophilic airway inflammation. This drug has been also shown to improve pulmonary function and symptoms in steroid-dependent asthma, resulting in a decrease in doses of inhaled corticosteroid. However, the precise mechanism by which suplatast tosilate exerts an antiasthmatic effect in vivo remains to be known. Our previous study showed the polarization of circulating type 1 dendritic cells (DC1)/type 2 dendritic cells (DC2) balance toward DC2 in asthma, which might be associated with its Th2-dominant immune response. In the present study, we attempted to clarify the effect of suplatast tosilate on DC1/DC2 balance in asthma. Using multicolor flow cytometry, we enumerated circulating DC1 and DC2 before and 8 weeks after treatment with suplatast tosilate in nine patients with asthma. Before the treatment, the patients with asthma had a significant higher percentage of DC2 together with a significant lower ratio of DC1/DC2 compared with normal subjects. Administration of suplatast tosilate significantly decreased the percentage of DC2, but not that of DC1, resulting in a significant raises of the ratio of DC1/DC2. Concomitantly, intracellular cytokine analysis showed that the percentage of IL-4 producing CD4⁺ T cells was significantly decreased after the treatment. These data suggest that suplatast tosilate normalizes the polarized DC1/DC2 balance toward DC2 in asthma, which may also alter its Th2-dominant Th1/Th2 balance toward Th1.     

Suplatast Tosilate Alters DC1/DC2 Balance in Peripheral Blood in Bronchial Asthma

Suplatast tosilate is an antiallergic drug that selectively suppresses Th2-cytokine production and inhibits airway hyperresponsiveness and eosinophilic airway inflammation. This drug has been also shown to improve pulmonary function and symptoms in steroid-dependent asthma, resulting in a decrease in doses of inhaled corticosteroid. However, the precise mechanism by which suplatast tosilate exerts an antiasthmatic effect in vivo remains to be known. Our previous study showed the polarization of circulating type 1 dendritic cells (DC1)/type 2 dendritic cells (DC2) balance toward DC2 in asthma, which might be associated with its Th2-dominant immune response. In the present study, we attempted to clarify the effect of suplatast tosilate on DC1/DC2 balance in asthma. Using multicolor flow cytometry, we enumerated circulating DC1 and DC2 before and 8 weeks after treatment with suplatast tosilate in nine patients with asthma. Before the treatment, the patients with asthma had a significant higher percentage of DC2 together with a significant lower ratio of DC1/DC2 compared with normal subjects. Administration of suplatast tosilate significantly decreased the percentage of DC2, but not that of DC1, resulting in a significant raises of the ratio of DC1/DC2. Concomitantly, intracellular cytokine analysis showed that the percentage of IL-4 producing CD4⁺ T cells was significantly decreased after the treatment. These data suggest that suplatast tosilate normalizes the polarized DC1/DC2 balance toward DC2 in asthma, which may also alter its Th2-dominant Th1/Th2 balance toward Th1.     

Effect of Azithromycin on the Severity of Bronchial Hyperresponsiveness in Patients with Mild Asthma

The effect of azithromycin on bronchial hyperresponsiveness was measured in a group of 11 patients with mild asthma. Azithromycin 250 mg orally was administered intermittently to all the patients twice a week for eight weeks. The only other treatment was inhaled β2 agonist, when required. A histamine inhalation test was performed at the beginning and at the fourth and the eighth week of the study. The mean PC₂₀ values increased significantly over the initial value at the eighth week after the administration of azithromycin (p < 0.05) but mean values for FEV1 and FEV1 percent predicted did not differ significantly. These results suggested that eight weeks of intermittent, low-dose administration of azithromycin in patients with mild asthma might reduce the severity of bronchial hyperresponsiveness.     

Peripheral Blood Th1 and Th2 Profile in Patients with Moderate Asthma: Effect of Inhaled Corticosteroid

The peripheral blood from healthy subjects and asthma patients was stimulated with phorbol 12-myristate 13-acetate and ionomycin, and the cells were stained with anti-CD4 antibody, permeabilized, stained with anti-IFN-γ and anti-IL-4 antibodies, and analyzed by flow cytometry. Compared with healthy subjects, asthma patients showed a greater percentage of both IL-4(+) IFN-γ (-) CD4 cells (Th2 cells) and IFN-γ(+) IL-4(-) CD4 cells (Th1 cells). The percentage of Th2 cells was correlated with serum IgE level. After treatment with inhaled corticosteroid, Th2 cells decreased at week 24, but not week 4. Long-term therapy with inhaled steroid may thus be required for improvement in lymphocytic inflammation.     

Renin-Angiotensin System Component Gene Polymorphism in Japanese Bronchial Asthma Patients

The influence of renin-angiotensin system (RAS) component gene polymorphism in the pathogenesis of bronchial asthma was investigated in an association study involving 119 bronchial asthma patients and 208 control subjects. The selected RAS polymorphisms were angiotensinogen (Agt) T235/M235 and angiotensin l-converting enzyme (ACE) insertion/deletion (l/D). The control allelic frequencies of the Agt T235/M235 (0.84/0.16) and ACE l/D (0.63/0.37) in this study were similar to the previous reports in Japanese normal population. The allelic frequencies of the Agt T235/M235 (0.84/0.16) and ACE l/D (0.65/ 0.35) among the asthma patients were not significantly different from those among the control subjects. There was no association between severity of bronchial asthma and the selected RAS component gene polymorphism. From these data, we conclude that in the Japanese population, the RAS component gene polymorphism is not associated with increased risk for bronchial asthma.     

不同潮气量通气对内毒素肺损伤大鼠Th1/Th2平衡的影响

目的 探讨内毒素肺损伤Thl/Th2平衡变化及不同潮气量通气对内毒素肺损伤大鼠Thl/Th2平衡的影响。方法 健康大鼠32只随机分成4组：生理盐水正常对照A组、内毒素肺损伤非机械通气B组、内毒素肺损伤＋小潮气量C组、内毒素肺损伤＋传统潮气量D组。监测平均动脉压(MAP)、心率(HR)，计算氧合指数（OI），检测PBMC中IFN-γ、IL-4浓度水平并计算IFN-γ/IL-4。取右肺中叶计算肺组织湿干比值(W/D)；观察右肺下叶肺组织病理变化；分析支气管肺泡灌洗液(BALF)中白细胞(WBC)与中性粒细胞(PMN)。结果 1. B、C、D组OI、W/D及病理学变化符合急性肺损伤表现；2. B组IFN-γ/IL-4比值明显高于A组；C组IFN-γ/IL-4比值明显低于B组；D组IFN-γ/IL-4比值明显高于B组；3. B组肺W/D及BALF中WBC、PMN计数明显高于A组；C组肺W/D及BALF中WBC、PMN计数明显低于B组；D组肺W/D及BALF中WBC、PMN计数明显高于B组。结论 1.注射LPS可成功复制大鼠急性肺损伤动物模型；2.内毒素致大鼠肺损伤早期，其PBMC中IFN-γ和IL-4浓度增加，IFN-γ/IL-4升高，Thl/Th2失衡；3.传统潮气量机械通气可加重内毒素肺损伤大鼠炎症反应，使肺损伤早期大鼠Th1/Th2失衡加剧；4.保护性通气策略的小潮气量机械通气可减轻内毒素肺损伤大鼠PMN在肺内“扣押”，改善Th1/Th2失衡状态。     

肝移植术后急性排斥反应中Th1/Th2变化的研究

目的探讨肝移植术后患者Th1/Th2在急性排斥反应中的变化及意义。方法采用双色流式细胞术分析21例肝移植患者手术前后外周血中的Th1细胞和Th2细胞的百分率,并计算Th1/Th2的变化。结果肝移植术后急性排斥过程中Th1细胞较术前有显著性升高,Th2细胞较术前无显著性变化,Th1/Th2较术前有显著性升高。排斥组与非排斥组相比,Th1/Th2有显著性升高。急性排斥时及排斥前Th1/Th2与术前相比有显著性差异。结论Th1/Th2的变化与肝移植急性排斥反应的发生密切相关。术后监测外周血中Th1/Th2是进行免疫监测及指导免疫抑制剂应用的简便有效的方法