白介素13在支气管哮喘发病中的作用

通过对支气管哮喘发病机制研究总结,对白介素13（IL-13）在支气管哮喘发病机制中的作用进行分析,特别是IL-13在免疫-炎症、气道重塑、基因遗传方面的作用进行阐述分析。     

白介素-13、白介素-18和肿瘤坏死因子-α血清浓度的变化在支气管哮喘发病中的作用

目的探讨白介素-13(IL-13)、白介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)在支气管哮喘发病机制中的作用。方法用酶联免疫吸附法(ELISA)测定48例支气管哮喘急性发作期治疗前后患者和40例健康对照者的血清IL-13、IL-18、TNF-α的浓度。结果(1)支气管哮喘急性发作期患者的血清IL-13、IL-18、TNF-α浓度显著高于正常对照组;(2)支气管哮喘患者急性发作期经肾上腺糖皮质激素等治疗病情缓解后IL-18、TNF-α血清浓度比较有显著性意义。结论支气管哮喘急性发作期患者的血清IL-13、IL-18、TNF-α的浓度高于健康对照组。急性发作期支气管哮喘患者经治疗病情缓解前后上述细胞因子浓度比较差异有显著性意义,提示IL-13、IL-18、TNF-α3种细胞因子参与支气管哮喘发病的免疫病理机制。     

三氧化二砷对支气管哮喘小鼠Th17细胞的影响

目的 观察三氧化二砷（ATO）对支气管哮喘小鼠Th 17细胞和IL-17的影响.方法 30只BALB/c小鼠随机分为对照组、哮喘组和ATO组.末次激发24小时后,测定气道肺阻力,收集支气管肺泡灌洗液（BALF）,细胞分类计数,流式细胞仪检测脾脏Th17细胞阳性率.ATO干预20小时后,检测培养上清液及BALF中IL-17及IL-4水平.结果 ①哮喘组BALF中各类炎症细胞均明显高于对照组（P＜0.05）,而ATO组明显低于哮喘组（P＜0.01）.②哮喘组肺阻力、脾脏CD4＋T细胞总数、Th17细胞阳性率、BALF中IL-17及IL-4均明显高于对照组,而ATO组上述指标均明显低于哮喘组（P＜0.05）.③ATO干预组培养上清液中IL-17、IL-4含量显著低于原液组（P＜0.01）.④Th17细胞阳性率、IL-17含量均与中性粒细胞数量呈显著正相关（P＜0.05）.结论 ATO能够抑制Th17细胞、IL-17的表达,减轻哮喘小鼠气道炎症及高反应性.     

雷公藤甲素对哮喘小鼠白介素8及白介素10表达的影响

目的研究雷公藤甲素对哮喘小鼠的治疗作用及对白介素8（IL-8）及白介素10（IL-10）表达的影响。方法 60只小鼠,随机分6组,每组各10只。分别为正常对照组（A）、哮喘组（B）、地塞米松治疗组（C）及不同剂量雷公藤甲素治疗组（D、E、F）,建立小鼠卵蛋白哮喘模型,用ELISA测定肺组织匀浆及气管肺泡灌洗液（BALF）中IL-8、IL-10表达水平。结果哮喘组小鼠肺组织匀浆及BALF中IL-8表达均高于正常对照组（P〈0.01）,哮喘组小鼠肺组织匀浆及BALF中IL-10表达均低于正常对照组（P〈0.01）,雷公藤甲素治疗组及地塞米松组IL-8低于哮喘组（P〈0.01）,雷公藤甲素治疗组及地塞米松组IL-10高于哮喘组（P〈0.01）。结论雷公藤甲素对哮喘小鼠有明显治疗作用,可抑制IL-8表达及提高IL-10的表达。     

红霉素抑制人支气管上皮细胞白介素-8基因的表达

目的 从分子水平上探讨红霉素(EM)的抗炎作用机制,为临床长期应用小剂量红霉素治疗慢性气道感染性疾病提供理论依据.方法 体外培养人支气管上皮细胞(16HBE),将细胞随机分为8组,先加入红霉素干预,后加入肿瘤坏死因子(TNF-α)刺激.分组如下:(1)空白对照组;(2)TNF-α(20ng/mL,16h);(3)EM(0.3μg/mL,24h)+TNF-α(20ng/mL,16h);(4)EM(3μg/mL,24h)+TNF-α(20ng/mL,16h);(5)EM(30μg/mL,24h)+TNF-α(20ng/mL,16h);(6)EM(0.3μg/mL,48h)+TNF-α(20ng/mL,16h);(7)EM(3μg/mL,48h)+TNF-α(20ng/mL,16h);(8)EM(30μg/mL,48h)+TNF-α(20ng/mL,16h).然后收集各组细胞分别提取RNA,应用逆转录聚合酶链反应(RT-PCR)方法检测白介素-8(IL-8)mRNA.结果 用前炎因子TNF-α刺激人支气管上皮细胞(16HBE)后,RT-PCR结果显示细胞IL-8mRNA表达增高.先加入不同浓度及作用时间的红霉素,后加入前炎因子TNF-α刺激人支气管上皮细胞(16HBE),RT-PCR结果显示各组细胞IL-8mRNA表达均降低,但其降低的水平与红霉素作用浓度及时间无明显关系.结论 前炎因子TNF-α能激活人支气管上皮细胞IL-8基因的表达,使其表达增高,促进炎症的发生发展,在炎症进程中发挥着重要作用.红霉素能抑制人支气管上皮细胞白介素-8基因表达,这可能是红霉素的抗炎作用机制之一.     

IL －4和 IL －13基因多态性与广东惠州地区儿童哮喘发生的相关性

目的：探讨IL－13（Arg130Gln）、IL－4（－589C／T）基因位点多态性与广东惠州地区儿童哮喘发生的相关性。方法采用PCR－RFLP法对34例广东籍哮喘患儿（哮喘组）及30例健康对照儿童（对照组）进行IL－13（Arg130Gln）、IL－4（－589C／T）位点的单核苷酸多态性检测。结果哮喘组与对照组IL－13（Arg130Gln）位点的基因型和等位基因频率差异有统计学意义（P＜0.05）;哮喘组与对照组IL－4（－589C／T）位点的基因型和等位基因频率差异无统计学意义（P＞0.05）;IL－13（Arg130 Gln）基因型AA及AG致儿童哮喘发生的相对危险度要高于基因型GG（P＜0.05）。结论 IL－13（Arg130Gln）基因多态性可能与广东惠州地区儿童哮喘发生存在相关性,并且基因型AA及AG对哮喘发生相对易感。     

成人支气管哮喘合并肺炎支原体感染对血清 IL－6、IL－13、IgE 水平的影响

目的：通过研究支气管哮喘发作合并肺炎支原体（MP）感染的成人患者血清中（IL －6）、IL －13及 IgE 的表达水平,探讨支原体感染与哮喘的发作及进展的关系。方法：收集2012年8月～2014年6月支气管哮喘发作并 MP 感染患者（年龄≥18岁）69例,非感染者73例,采用 ELISA 方法检测 IL －6、IL －13、IgE 水平。结果：IL －6、IL －13、IgE 水平在支气管哮喘合并 MP 感染发作组分别为（102.4±10.3 pg／ml）、（375.2±92.5 pg／ml）、（739.5±214.7U ／ml）,显著高于非 MP 感染组（62.7±7.4 pg／ml）、（129.8±31.6 pg／ml）、（398.7±123.2 U ／ml）,P ＜0.01。结论：肺炎支原体感染合并支气管哮喘患者 IL －6、IL －13、与 IgE 水平升高更加明显,表明 MP 感染是支气管哮喘发生发展过程中的重要危险因素。     

ADAM15在支气管哮喘表达及意义

目的:研究整合素金属蛋白酶15(a disintegrin and metalloproteinase 15,ADAM15)在支气管哮喘中表达及可能机制.方法:收集本院32例哮喘患者及同期16例健康对照的临床资料及血清标本,酶联免疫吸附法检测血清ADAM15水平,分析其与外周血嗜酸性粒细胞数、血清总IgE相关性.构建O...     

早期T细胞激活过程中NF-κB活化的研究

目的:研究T细胞在CD3/CD28双信号通路共刺激活化早期,细胞中NF-κB的活化以及三氧化二砷对该活化作用的影响。方法:用CD3/CD28单克隆抗体(浓度分别为10μg/ml和5μg/ml)双信号通路对Hut-78细胞进行共刺激30min,采用TransAM的方法观察Hut-78细胞内p65的表达状况,并研究三氧化二砷对该表达的影响。结果:Hut-78细胞在CD3/CD28双信号通路共刺激30min后,细胞核中p65的表达显著下降(P<0.05)。三氧化二砷组p65表达要高于单纯CD3/CD28作用组。全细胞蛋白提取液中的p65表达水平的趋势与细胞核提取液类似。结论:T细胞经过CD3/CD28共刺激,细胞内NF-κB不能得到迅速活化,反而有下降的趋势。三氧化二砷能够促进NF-κB的早期活化。     

Inhibition of interleukin-13 gene expression in T cells through GATA-3 pathway by arsenic trioxide

Aarsenic trioxide （AT） has a long history of use in both traditional Chinese medicine and in modem medicine in asthma therapy. Recently, Yin et al＇ found that AT even at small doses reduced the airway inflammation of sensitized guinea pigs. However the mechanism underlying this is still largely unknown. Interleukin 13 （IL-13）, as one of the important TH2 cytokines, plays an important role in asthma pathogenesis through promoting eosinophilic inflammation,     

Arsenic trioxide inhibits p-glycoprotein expression in multidrug-resistant human leukemia cells that overexpress the MDR1 gene

Objective To investigate the effects of arsenic trioxide (As2O3) on the apoptosis and p-glycoprotein (P-gp) expression of multidrug-resistant human leukemia cells.Methods Human multidrug-resistant leukemia cell line K562/ADM overexpressing the MDR1 gene, was used as the target cells. The cell proliferating activity was assessed using the MTT colorimetric assay. Cytomorphology was investigated under light, confocal and electron microscopes. DNA fragmentation was examined using agarose gel electrophoresis, while p-gp expression, cell cycle status and sub-G1 cells were determined using flow cytometry.Results Zero point five to 20 μmol/L As2O3 inhibited the proliferation of K562/ADM cells, and K562/ADM cells were more sensitive to As2O3 than the parental K562 cells. As2O3-induced apoptosis of K562/ADM cells was determined by the observance of typical morphological changes and the appearance of DNA ladder and sub-G1 cell populations. As2O3 significantly inhibited the P-gp expression of K562/ADM cells, and synergistically enhanced the sensitivity of the drug-resistant cells to adriamycin.Conclusions As2O3 induces growth-inhibition and apoptosis, down-regulates P-gp expression and exerts a synergistic effect in combination with adriamycin in multidrug-resistant leukemia cells.     

Arsenic Trioxide Inhibits Proliferation in K562 Cells by Changing Cell Cycle and Survivin Expression

To study the mechanisms involved in the inhibition of chronic myeloid leukemic cells (K562) proliferation induced by arsenic trioxide (As2O3) and to explore the potential role of Survivin, an inhibitor of apoptosis protein, in the regulation of As203 induced cell apoptosis, K562 cells were cultured with As203 of different concentrations. Cells were collected for proliferation analysis by MTT assay. Cell cycle distribution and cell apoptosis were analyzed by flow cytometry. Expression of Survivin protein and mRNA were detected by flow cytometry and RT-PCR, respectively. Our results showed that As2O3 (2-10/μmol/L) inhibited K562 cells growth effectively, but it did not induce cells apoptosis significantly. The percentage of K562 cells at G2/M phase increased in proportion to As2O3 concentrations, and the expression of Survivin mRNA and content of Survivin protein was up-regulated accordingly. It is concluded that As2O3 inhibited K562 cells growth by inducing cell cycle arrest mainly at G2/M phase. Over-expression of Survivin gene and protein might be one of the possible mechanisms contributing to K562 cells‘ resistance to As2O3-induced apoptosis.     

Arsenic trioxide up-regulates Fas expression in human osteosarcoma cells

Background Osteosarcoma is a common primary malignant tumor of bone with a poor prognosis due to its propensity for metastasis. The prognosis of patients is highly dependent on the presence or absence of lung metastasis and on the effectiveness of treatment against it. It has been reported that low level expression of Fas protein in human osteosarcoma cell is closely associated with lung metastasis. A large number of studies have shown that arsenic trioxide （ATO） can inhibit proliferation and induce apoptosis of many cancer cell lines; however, its effects on human osteosarcoma cells （Saos-2 cell line） remains unknown. The aim of this study was to investigate the effects of ATO on Saos-2 cells and to characterize its mechanism of Fas-expressing. Methods A group of Saos-2 cells was treated with or without 0.5, 1, 2, 4 and 8 pmol/L ATO for three successive days, and the cytotoxicity of ATO was determined by an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide （MTT） assay. Morphological changes in cells were studied by acridine orange/ethidium bromide （AO/EB） double staining. Flow cytometry （FCM） was used to assay cell DNA distribution. Another group of cells was pretreated with 10 nmol/L matrix metalloproteinase 7 （MMP-7） for 3 hours. They were then incubated with or without 2 pmol/L ATO for 24, 48 and 72 hours. Cytotoxicity, Fas protein and mRNA levels were systematically studied using MTT, Western blotting and real-time PCR, respectively. Cell proliferation, cell cycle progression and apoptosis were examined in this study. Results Proliferation of Saos-2 cells was inhibited by ATO in both a dose- and time-dependent manner. The IC50 values at 24, 48 and 72 hours were 9.30, 5.54 and 3.49 pmol/L, respectively. The survival rate of Saos-2 cells in the MMP-7 and ATO co-treated group was significantly higher than the ATO group, but it was lower than the control group. ATO induced G1 phase arrest of the cell cycle and very efficiently stimulated apoptosis in Saos-2 cells, as evidenced by flow cytometric detection of sub-G1 DNA content and AO/EB staining. Western blotting results indicated that Fas （FasL） protein expression in osteosarcoma cultures markedly increases in a time dependent manner after exposure to ATO. Compared with control, treatment with ATO 2 IJmol/L and 4 pmol/L for 48 hours, resulted in increase of Fas gene expression to 28.31% and 56.74%, respectively. Our results indicated that ATO induced-apoptosis of Saos-2 cells may be mediated through the Fas pathway. Conclusions ATO suppressed cell proliferation of Saos-2 cell in a dose- and time-dependent manner and increased Fas protein expression. However, Fas-mediated apoptosis was incompletely interrupted by MMP-7, which suggested that other molecular mechanisms may mediate this process.     

槐杞黄对哮喘气道及血清IL-13水平的影响

目的:探讨IL-13在哮喘患儿血清中的表达及槐杞黄对于哮喘大鼠IL-13表达的可能影响及与布地奈德的共同作用?方法:选择 10例哮喘缓解期儿童 (哮喘组)及10例健康儿童 (健康对照组)为研究对象?两组年龄?性别?体质指数比较差异均无统计学意义?测试两组儿童初诊时肺功能,并利用酶联免疫吸附试验(enzymelinkedimmunosorbentassay,ELISA)检测两组儿童血清 IL-13的水平?取50只4~6周龄的健康雄性SD大鼠,适应性喂养1周后,随机分为5组,即对照组?哮喘模型组?布地奈德组?槐杞黄组?布地奈德联合槐杞黄组?用卵清白蛋白(ovalbumin,OVA)致敏及激发共持续6周,建立哮喘模型?通过HE染色观察肺部炎症细胞浸润情况?应用ELISA检测及比较各组大鼠之间肺泡灌洗液(bronchoalveolar lavage fluid,BALF)及血清中IL-13浓度的差异?结果:与健康对照组儿童比较,哮喘组儿童初诊时第1秒用力呼气量占预测值百分比及呼气峰流速占预测值百分比均升高,血清IL-13水平升高(P < 0.05)?大鼠肺组织病理学改变:模型组气道周围大量炎症细胞浸润?柱状细胞增生?气管壁增厚;对照组则无上述表现;相比模型组,布地奈德组?槐杞黄组?布地奈德联合槐杞黄组大鼠不同程度减轻(P < 0.05)?模型组BALF及血清中IL-13的浓度增加(P < 0.05)?相较于模型组,布地奈德组?槐杞黄组?布地奈德联合槐杞黄组大鼠BALF及血清IL-13浓度有不同程度降低(P < 0.05),且布地奈德联合槐杞黄组变化比布地奈德组更明显(P < 0.05)? 结论:在OVA诱导的大鼠哮喘模型中,使用布地奈德?槐杞黄或是布地奈德联合槐杞黄治疗可以不同程度改善气道IL-13分泌,因而可能改善气道炎症?布地奈德联合槐杞黄治疗可能存在协同效应?     

IL-13基因多态性位点rs20541和rs1800925单核苷酸多态性 与儿童支气管哮喘关联性的研究

目的:研究IL-13基因多态性位点rs20541和rs1800925单核苷酸多态性与儿童支气管哮喘发病的关联性.方法:应用聚合酶链反应(PCR)、等位基因特异性PCR(AS-PCR)和直接测序方法检测105例儿童哮喘患者和112名正常儿童的IL-13基因上rs20541和rs1800925单核苷酸多态性分布特点.结果:第4外显子中的+2044G→A(rs20541)哮喘组与正常对照组呈现明显差异(P<0.05).而-1112C→T(rs1800925)位点两组间差异无统计学意义(P>0.05).结论:+2044G→A(rs20541)与哮喘患儿的发病具有强烈相关性.-1112C→T无明显相关性.     

Arsenic trioxide downregulates the expression of annexin II in bone marrow cells from patients with acute myelogenous leukemia

Background Most patients with acute myelogenous leukemia （AML） suffer from disordered hemostasis. We have previously shown that annexin II （Ann II）, a high-affinity co-receptor for plasminogen/tissue plasminogen activator, plays a central role in primary hyperfibrinolysis in patients with acute promyelocytic leukemia （APL）. The expression of Ann II in cells from patients with major subtypes of AML and the effect of arsenic trioxide （As203） on Ann II expression in AML cells were investigated to determine whether As203-mediated downregulation of Ann II could restore hemostatic stability. Methods A total of 103 patients （48 females and 55 males; age, 19-58 years） were included. Plasma samples were collected before and after treatment as well as after complete remission. Ann II and plasminogen activation were measured in leukemic cells during treatment with 1 pmol/L As203. Results Before AS203 treatment, Ann II mRNA expression （real-time PCR） was the highest in M3 cells （P 〈0.05）, higher in M5 cells than that in M1, M2, M4, and M6 cells （P〈0.001）, and positively correlated with Ann II protein expression （flow cytometry） （r=0.752, P 〈0.01）. Exposure for up to 120 hours to As203 （1 μmol/L） had no significant effect on Ann II protein in M1 and M2 leukemic cells, but decreased Ann II protein expression twofold within 48 hours of exposure in M3 cells （P 〈0.05） and twofold within 96 hours in M5 cells （P 〈0.05）. The rate of plasmin generation was higher in APL, M5, and M4 cells than in M1, M2, and M6 cells. Conclusions As203 may reduce hyperfibrinolysis in AML by downregulation of Ann 11. Furthermore, As2O3 affects more than one form of AML （APL, M4 and M5）, suggesting its potential role in their management.     

Arsenic trioxide in treatment of de novo acute basophilic leukemia

Acute basophilic leukemia (ABL) is a rare subtype of acute myeloid leukemia (AML),accounting for 4％-5％ of AML and less than 2％ of all hematopoietic malignancies.It is usually characterized by a very rapid clinical course,symptoms of hyperhistaminemia,peptic ulceration,gastrointestinal cerebrovascular bleeding and resistance to therapy.1 However,the clinical outcome of ABL remains disappointing.Most patients died within 1 year after diagnosis.We reported a de novo ABL case in a 70-year-old patient treated with single-agent arsenic trioxide (ATO).And prolonged survival was observed.