白藜芦醇苷对内毒素休克肺损伤大鼠Th1/Th2失衡的干预作用

目的:探讨白藜芦醇苷对内毒素休克肺损伤大鼠Th1/Th2类细胞因子水平失衡的影响。方法:采用颈静脉注入LPS复制大鼠急性肺损伤模型,ELISA法检测其外周血单个核细胞产生IFN-γ、IL-4水平及IFN-γ/IL-4比值并观察PD对其影响。结果:在体内实验中,PD能使IL-4水平显著升高(P<0.001),使失衡的IFN-γ/IL-4比值接近正常对照组,肺组织病理切片显示肺损伤减轻;在体外实验中,与急性肺损伤对照组比较,IFN-γ、IL-4水平虽无显著差异(P>0.05),但IFN-γ/IL-4比值接近正常对照组(P>0.05)。结论:PD在体内、外实验中,能纠正内毒素休克大鼠体内失衡的Th1/Th2类细胞因子水平,减轻肺损伤。     

N-acetylcysteine abrogates acute lung injury induced by endotoxin

1. Acute lung injury (ALI) or acute respiratory distress syndrome is a serious clinical problem with high mortality. N-Acetylcysteine (NAC) is an anti-oxidant and a free radical scavenger. It has been reported recently that NAC ameliorates organ damage induced by endotoxin (lipopolysaccharide; LPS) in conscious rats. The present study was designed to evaluate the effects of NAC on LPS-induced ALI and other changes in anaesthetized rats. 2. Sprague-Dawley rats were anaesthetized with pentobarbital (40 mg/kg, i.p.). Endotracheal intubation was performed to provide artificial ventilation. Arterial pressure and heart rate were monitored. The extent of ALI was evaluated with the lung weight (LW)/bodyweight ratio, LW gain, exhaled nitric oxide (NO) and protein concentration in bronchoalveolar lavage (PCBAL). Haematocrit, white blood cells, plasma nitrate/nitrite, methyl guanidine (MG), tumour necrosis factor (TNF)-alpha and interleukin (IL)-1b were measured. Pathological changes in the lung were examined and evaluated. 3. Endotoxaemia was produced by injection of 10 mg/kg, i.v., LPS (Escherichia coli). Animals were randomly divided into three groups. In the vehicle group, rats received an i.v. drip of physiological saline solution (PSS) at a rate of 0.3 mL/h. The LPS group received an i.v. drip of PSS for 1 h, followed by LPS (10 mg/kg by slow blous injection, i.v., over 1-2 min). Rats in the LPS + NAC group received NAC by i.v. drip at a rate of 150 mg/kg per h (0.3 mL/h) for 60 min starting 10 min before LPS administration (10 mg/kg by slow blous injection, i.v., over 1-2 min). Each group was observed for a period of 6 h. 4. N-Acetylcysteine treatment improved the LPS-induced hypotension and leukocytopenia. It also reduced the extent of ALI, as evidenced by reductions in LW changes, exhaled NO, PCBAL and lung pathology. In addition, NAC diminished the LPS-induced increases in nitrate/nitrite, MG, TNF-a and IL-1b. 5. In another series of experiments, LPS increased the mortality rate compared with the vehicle group (i.v. drip of PSS at a rate of 0.3 mL/h) during a 6 h observation period. N-Acetylcysteine, given 10 min prior to LPS, significantly increased the survival rate. 6. The results of the present study suggest that NAC exerts a protective effect on the LPS-induced ALI. The mechanisms of action may be mediated through the reduction of the production of NO, free radicals and pro-inflammatory cytokines.     

呼吸道合胞病毒(RSV)与Th1/Th2细胞因子的关系研究进展

目的对感染呼吸道合胞病毒产生的Th1、Th2型细胞因子种类及其对Th1/Th2型反应平衡的影响进行综述。方法在查阅国内外文献的基础上,介绍各种细胞因子在呼吸道合胞病毒(RSV)感染中的作用、地位。结果与结论虽然到目前为止各种细胞因子在各实验中得出的结果,甚者是对于RSV感染更倾向于发生Th1型还是Th2型反应仍有争论,但Th1/Th2反应平衡在RSV治疗和开发防治药物方面仍具有重要意义。     

右美托咪定激活PI3K/Akt通路改善LPS诱导的大鼠急性肺损伤实验研究

目的 考察右美托咪定（Dex）对脂多糖（LPS）诱导的大鼠急性肺损伤（ALI）的影响,并探讨可能与PI3K/Akt信号通路改变的相关机制。方法 将80只SD大鼠根据建模方法随机分为5组：对照组、模型组、Dex低剂量组、Dex高剂量组、Dex+LY294002组,建模24 h时处死并分离肺组织,观察组织病理切片,检查肺水肿程度及微血管通透性,采用Western blotting法检测Akt及其磷酸化蛋白以及线粒体凋亡信号相关蛋白的表达水平,检查线粒体细胞色素C、膜电位及ATP含量,并用电镜观察线粒体形态,采用流式细胞术检测肺组织细胞凋亡情况,采用DCFH-DA探针荧光显微镜法检测细胞内的活性氧（ROS）水平。结果 LPS滴注24 h后,病理切片表现有明显的ALI特征,Dex在50 μg/kg剂量时可明显改善LPS诱导的ALI病理特征;以对照组为参照,p-Akt（Thr308）与p-Akt（Ser473）的相对表达量模型组未发生明显变化,Dex组则均显著增高（P<0.001）,在Dex+LY294002组的表达水平较Dex组显著降低（P<0.001）;模型组的Bax与Bcl-2蛋白较对照组的相对表达量分别显著升高与降低（P<0.001）,Dex组分别降低与升高至接近对照组水平,Dex+LY294002组又分别回升与回降;对照组、模型组、Dex组、Dex+LY294002组的肺组织细胞凋亡率分别为2.4%、19.5%、6.7%、13.1%;各组的ROS水平变化与肺组织细胞凋亡情况一致性良好。结论 Dex可通过激活PI3K/Akt信号通路减轻LPS诱导的肺组织细胞凋亡及线粒体凋亡信号激活,继而改善ALI,发挥肺保护作用。     

屈头鸡果对内毒素致小鼠急性肺损伤的保护作用

目的 研究屈头鸡果在内毒素致小鼠急性肺损伤中的作用及其机制.方法 将40只SPF级雄性BALB/c小鼠分为正常组、对照组、模型组和实验组,每组各10只.经鼻滴入20 mg·kg-1脂多糖构建内毒素致小鼠急性肺损伤模型,其中对照组和实验组分别灌胃屈头鸡果2.16 g·kg-1,正常组和模型组灌胃等量生理盐水.观察各组小鼠...     

1型糖尿病外周血Th1/Th2细胞因子表达水平的研究

目的检测1型糖尿病患者外周血CD_4~+T细胞分泌细胞因子的水平变化,探讨患者Th1/Th2细胞因子的平衡状态及其在1型糖尿病中的作用。方法对49例1型糖尿病患者和30例健康时照组外周血用刺激物刺激细胞,增加细胞内细胞因子的表达,再加入荧光标记的特异性抗细胞因子单克隆抗体,特异性抗原抗体结合,以流式细胞仪分析特异性细胞因子表达水平。结果1型糖尿病患者Th1型细胞因子干扰素-γ(IFN-γ)、白细胞介素-2(IL-2)表达水平较正常对照组升高,差异有统计学意义(P<0.01)。Th2型细胞因子白细胞介素-4(IL-4)、白细胞介素-10(IL-10)表达水平较正常时照组显著降低,差异有统计学意义(P<0.01)。结论1型糖尿病患者Th1/Th2平衡失调,Th1型反应模式处于优势状态,Th2型反应模式处于弱势状态。Th1/ Th2平衡向Th1方向漂移。     

紫杉醇化疗方案对乳腺癌患者血清Th1/Th2细胞因子水平的影响

目的探讨紫杉醇化疗方案对乳腺癌患者血清Th1/Th2细胞因子水平的影响。方法将100例乳腺癌术后辅助化疗患者随机(1︰1)分为2组,常规组行AC(阿霉素-环磷酰胺,每3周1次)-P(紫杉醇80mg·m~(-2),每周1次)方案化疗,剂量密集组,行AC(阿霉素-环磷酰胺每2周1次)-P(紫杉醇175mg·m~(-2),每2周1次)方案化疗,化疗前后观察患者血清细胞因子水平变化。结果常规组及剂量密集组化疗后患者血清IL-2、TNF-α及IFN-γ浓度均显著增高(P<0.05),血清IL-4、IL-6及IL-10水平显著降低(P<0.05)。结论常规组及剂量密集组均能够改善乳腺癌患者细胞免疫功能;剂量密集组较常规组具有更强的恢复细胞免疫为主的抗肿瘤免疫能力,且未显著增加毒性和不良反应。     

康艾注射液对人结直肠癌Lovo细胞Th1/Th2类细胞因子的影响

Objective To explore the effect of Kang'ai injection on Th1/Th2 cytokine in human colon carcinoma cell line Lovo.Methods Lovo cells were cultured by conventional ways until they were in exponential growth phase and then divided into 4 groups:one control group and three experimental groups with different concentration of Kang'ai injection.After 72 hours culture,Th1 type cytokines (IL-2,IFN-γ) and Th2.type cytokines (IL-4,IL-10) in the culture supernatant of Lovo cells in each group were separately detected by double-antibody sandwich enzyme-linked immunosorhent assay (ELISA).Results After the coculture of Lovo cells and Kang'ai injection,Th2 type cytokine levels in the supernatants were significant lower than the control group (P＜0.05),and Th1 type cytokines in each group were of little change.Conclusions Kang'ai injection markedly downregulated Th2 type cytokines in Lovo cells,and contributed to the switching from Th2 to Th1.     

Maresin 1 mitigates LPS-induced acute lung injury in mice

BACKGROUND AND PURPOSE

Acute lung injury (ALI) is a severe illness with a high rate of mortality. Maresin 1 (MaR1) was recently reported to regulate inflammatory responses. We used a LPS-induced ALI model to determine whether MaR1 can mitigate lung injury.

EXPERIMENTAL APPROACH

Male BALB/c mice were injected, intratracheally, with either LPS (3 mg·kg⁻¹) or normal saline (1.5 mL·kg⁻¹). After this, normal saline, a low dose of MaR1 (0.1 ng per mouse) or a high dose of MaR1 (1 ng per mouse) was given i.v. Lung injury was evaluated by detecting arterial blood gas, pathohistological examination, pulmonary oedema, inflammatory cell infiltration, inflammatory cytokines in the bronchoalveolar lavage fluid and neutrophil–platelet interactions.

KEY RESULTS

The high dose of MaR1 significantly inhibited LPS-induced ALI by restoring oxygenation, attenuating pulmonary oedema and mitigating pathohistological changes. A combination of elisa and immunohistochemistry showed that high-dose MaR1 attenuated LPS-induced increases in pro-inflammatory cytokines (TNF-α, IL-1β and IL-6), chemokines [keratinocyte chemokine, monocyte chemoattractant protein-5, macrophage inflammatory protein (MIP)-1α and MIP-1γ], pulmonary myeloperoxidase activity and neutrophil infiltration in the lung tissues. Consistent with these observations, flow cytometry and Western blotting indicated that MaR1 down-regulated LPS-induced neutrophil adhesions and suppressed the expression of intercellular adhesion molecule (ICAM)-1, P-selection and CD24.

CONCLUSIONS AND IMPLICATIONS

High-dose MaR1 mitigated LPS-induced lung injury in mice by inhibiting neutrophil adhesions and decreasing the levels of pro-inflammatory cytokines.     

N-乙酰半胱氨酸对大鼠急性脊髓损伤后继发性脊髓损伤的保护作用

目的 探讨N-乙酰半胱氨酸(NAC)对大鼠脊髓急性损伤后继发性脊髓损伤的保护机制.方法 成年SD大鼠18只随机均分成单纯椎板切除(对照组)、急性脊髓损伤(损伤组)和急性脊髓损伤后NAC治疗(治疗组)三组.用30 g力量动脉瘤夹从两侧夹闭脊髓30 s建立脊髓损伤模型.治疗组术后15 min、1、2和3d分别予NAC 150 mg/kg腹腔注射;对照组和损伤组腹腔注射等量生理盐水.所有动物于术后3d处死取材,用凝胶电泳迁移率分析和免疫组化检测脊髓组织中核因子κB(NF-κB),ELISA法检测肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6表达.结果 损伤组脊髓组织中NF-κB、TNF-α、IL-1β和IL-6水平均比对照组升高(P＜0.05),治疗组脊髓组织中各因子水平均比损伤组显著降低(P＜0.05).结论 NAC可以通过抑制大鼠急性脊髓损伤后NF-κB的活性,进一步下调TNF-α、IL-1β和IL-6的表达,从而减轻继发性炎症反应所导致的脊髓损伤.     

Patulin influences the expression of Th1/Th2 cytokines by activated peripheral blood mononuclear cells and T cells through depletion of intracellular glutathione

Patulin is a mold toxin secreted mainly by fungi of the Penicillium species. Exposure generally results from consumption of moldy fruits and fruit products. Since recent studies identified mold exposure as a risk factor for allergic diseases, we examined the effects of patulin on human peripheral blood mononuclear cells (PBMC) prepared from buffy coats of healthy donors. Cells were stimulated with CD3- and CD28-specific antibodies in the presence or absence of patulin. Effects of patulin on PBMCs were evaluated by proliferation, viability assays, and cytokine ELISAs. The presence of 50 ng/mL patulin strongly decreased the amounts of several cytokines in the supernatant of stimulated PBMCs. This decrease in cytokine secretion was not due to cytotoxic effects of patulin. Moreover, the extent of the reduction of cytokine amounts was cytokine specific, affecting some (IL-4, IL-13, IFNgamma, and IL-10), but not others (IL-8, IL-5). We show that all effects could be abolished by adding thiol containing compounds. A depletion of intracellular GSH could be measured after incubation of cells with patulin. Taken together, our data indicate that patulin modulates the functional activation of PBMCs with respect to proliferation and cytokine secretion patterns by depletion of intracellular GSH. The depletion of intracellular glutathione may influence the balance between Th1 and Th2 cells and have implications for allergic diseases.     

康艾注射液对人结直肠癌Lovo细胞Th1/Th2类细胞因子的影响

目的 探讨康艾注射液对人结直肠癌Lovo细胞Th1/Th2状态的影响.方法 常规培养Lovo细胞,待细胞处于指数增长期再分成4组:1组为空白对照,另外3组加入不同浓度的康艾注射液.培养72h后,采用双抗夹心酶联免疫吸附法(ELISA)分别检测各组Lovo细胞培养上清中Th1类细胞因子(IL-2,IFN-γ)和Th2类细胞因子(IL-4,IL-10)的水平.结果 Lovo细胞与康艾注射液共培养后,上清液中Th2类细胞因子的含量较对照组明显降低(P＜0.05),而Th1类细胞因子在各组中的含量变化不大.结论康艾注射液可显著下调Lovo细胞Th2类细胞因子水平,具有促进其Th2向Th1逆转的倾向.     

血必净辅助治疗重症肺炎的疗效及对患者肾功能和血清Th1/Th2细胞因子的影响

目的 探讨血必净辅助治疗重症肺炎的疗效及对患者肾功能和血清Th1/Th2细胞因子的影响。方法 取2015年1月-2017年2月南阳医学高等专科学校第一附属医院诊治的196例重症肺炎患者,根据随机数字表法分为对照组和观察组各98例。均给予吸氧、化痰和营养支持等常规指标和抗生素治疗;观察组在此基础上给予血必净治疗,连续治疗1周。比较两组的疗效,治疗前后的肾功能指标水平以及血清Th1/Th2细胞因子水平。结果 观察组的治疗总有效率为94.90%,高于对照组的78.57%,差异有统计学意义（P < 0.05）。治疗前,两组的血肌酐（SCr）、血尿素氮（BUN）和内生肌酐清除率（Ccr）水平差异无统计学意义;治疗后,对照组的SCr、BUN水平略高于治疗前,Ccr水平略低于治疗前但不具统计学差异,而观察组的SCr、BUN水平高于治疗前,Ccr水平低于治疗前,差异有统计学意义（P < 0.05）;治疗后观察组的SCr、BUN水平高于对照组,且Ccr水平低于对照组,差异有统计学意义（P < 0.05）。治疗前,两组的干扰素-γ（INF-γ）和白细胞介素-4（IL-4）水平和INF-γ/IL-4差异无统计学意义;治疗后两组的INF-γ水平、INF-γ/IL-4高于治疗前,IL-4水平低于治疗前,同组治疗前后比较差异有统计学意义（P < 0.05）;且治疗后观察组的INF-γ水平、INF-γ/IL-4高于对照组,IL-4水平低于对照组,差异有统计学意义（P < 0.05）。结论 血必净联合抗生素治疗重症肺炎效果明显,有利于保护肾脏功能,调节血清Th1/Th2细胞因子水平,增强免疫力。     

CpG-ODN对哮喘小鼠Th1/Th2相关细胞因子表达的影响

目的：探讨CpG-ODN对哮喘小鼠Th1/Th2相关细胞因子的调节作用及气道炎症的抑制作用。方法：采用酶联免疫中附法（ELISA）分别检测CpG-ODN对哮喘小鼠抗原首次激发后24小时和抗末次激发后24小时血清及支气管肺泡灌洗液（BALF）中Th2细胞因子IL－4和Th1细胞因子IFNγ水平的影响，并与激素治疗组和哮喘组对照。结果：（1）抗原末次激发后24小时，CpG-ODN 血清及BALF中IFNγ水平高于哮喘组及地塞米松组（P＜0．001）。（2）抗原首次激发后24小时及抗原末次激发后24小时血清和BALF中IL－4水平低于哮喘组（P＜0．05），与地塞米松组比较无显著性差异（P＞0．05）。（3）抗原末次激发后24小时，CpG-ODN组外周血和BALF中嗜酸性粒细胞计数均低于哮喘组（P＜0．001）。结论：CpG-ODN不仅有具有与地塞米松相同的下调Th2细胞因子表达和抑制嗜酸性粒细胞的作用，而且还具有地塞米松所没有的诱导Th1细胞因子表达的作用。说明其可能通过调节Th1/Th2平衡而达到在早期抑制哮喘发病的目的。     

中药热毒宁对急性肺损伤兔肺内致炎因子的影响

目的:研究中药热毒宁(RDN)对于内毒素性急性肺损伤兔肺内致炎因子的影响。方法:静脉注射内毒素建立兔急性肺损伤的模型,动物共分3组:对照组,模型组和药物组。应用RDN预处理后,不同时间点检测外周血液白细胞介素8(IL-8)、白细胞介素10(IL-10)、肿瘤坏死因子(TNF-α)的含量,流式细胞仪测定外周血及灌洗液内的多形核白细胞(PMN)凋亡、坏死细胞比例及呼吸爆发功能。结果:模型组血IL-8、IL-10、TNF-α含量较对照组显著升高,灌洗液PMN成活比例增加,凋亡延迟,呼吸爆发明显增强(P＜0．05)。药物组上述变化明显受抑(P＜0．05,P＜0．01)。结论:RDN的预处理可抑制内毒素性兔急性肺损伤的发生与发展。     

类风湿关节炎患者外周血Th1、Th2和Th17细胞亚群表达比例变化的特点

目的研究类风湿关节炎（rheumatoidarthritis,RA）患者外周血中Th1、Th2和Th17细胞亚群表达比例变化,分析RA免疫学发病机制。方法运用流式细胞术检测RA患者及对照组外周血单个核细胞（PBMCs）中Th1、Th2、Th17细胞表达比例。并用酶联免疫吸附法检测RA患者及对照组血清中IFN-γ、IL-4、IL-17的浓度。结果RA患者CD4＋T细胞明显高于健康对照组（P〈0．05）;活动期RA患者Th1、Th17及其分泌的细胞因子含量明显高于健康对照组（P〈0．05）;非活动期RA患者Th17也明显高于健康对照组（P〈0．05）。结论类风湿关节炎Th1、Th2、Th17平衡改变与其疾病的活动度有关,处于活动期Th1、Th17反应增强,非活动期Th1、Th2、Th17趋于平衡。通过调节Th1、Th2、Th17平衡,有可能为RA的治疗提供一条新思路。