Macrolide antibiotics like azithromycin increase lipopolysaccharide-induced IL-8 production by human gingival fibroblasts

Objective

Macrolide antibiotics are reported to modulate the production of cytokines in various type of cells. We examined the effect of macrolide antibiotics on inflammatory cytokines (IL-6 and IL-8) and chemical mediator (PGE₂) and also matrix metalloproteinases (MMPs) productions by human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS).

Methods

The effect of macrolide antibiotics [erythromycin (EM), azithromycin (AZM) and josamycin (JOM)] on HGFs proliferation were examined by MTT assay. HGFs were treated with LPS from Porphyromonas gingivalis (PgLPS) and macrolide antibiotics, and IL-6, IL-8 and PGE₂ levels were evaluated by ELISA. MMPs were detected by gelatin zymography.

Results

AZM slightly but significantly decreased HGFs proliferation, while EM and JOM did not affected. AZM increased PgLPS-induced IL-8 production dose-dependently, while AZM did not alter IL-6 and PGE₂ productions. EM and JOM did not altered PgLPS-induced IL-6, IL-8 and PGE₂ productions. All macrolide antibiotics did not alter MMPs production. These results indicate that macrolide antibiotics have no direct anti-inflammatory effect. However, the use of the inhibitors of cell signaling pathway failed to reveal the mechanism that AZM enhanced PgLPS-induced IL-8 production.

Conclusion

These results suggest macrolide antibiotics have an indirect anti-inflammatory effect as a result of their antimicrobial properties. Because AZM increased LPS-induced IL-8 production by HGFs, the possibility is considered that neutrophils may be migrated to periodontal tissue and phagocytize the periodontopathic bacteria more efficiently.     

EFFECTS OF INTERLEUKIN－4 ON GRANULOCYTE－MACROPHAGE－COLONY FORMATION FROM MURINE BONE MARROW CELLS AND HEMATOPOIETIC RECONSTIT

ＥＦＦＥＣＴＳＯＦＩＮＴＥＲＬＥＵＫＩＮ－４ＯＮＧＲＡＮＵＬＯＣＹＴＥ－ＭＡＣＲＯＰＨＡＧＥ－ＣＯＬＯＮＹＦＯＲＭＡＴＩＯＮＦＲＯＭＭＵＲＩＮＥＢＯＮＥＭＡＲＲＯＷＣＥＬＬＳＡＮＤＨＥＭＡＴＯＰＯＩＥＴＩＣＲＥＣＯＮＳＴＩＴＵＴＩＯＮＦＯＬＬＯＷＩＮ...     

弥漫性泛细支气管炎给我们的启示

弥漫性泛细支气管炎是发现于日本的新呼吸系统疾患,与种族相关抗原关系密切,是一种遗传相关性疾病.十四、十五元环大环内酯类抗生素对其有特殊疗效.该病及大环内酯类药物疗法的发现过程,提示我们应重视疾病的人种差别,不要轻易放过医疗过程中遇到的偶然现象.     

体外实验中缺氧对中性粒细胞凋亡的影响

目的：通过在缺氧条件下体外培养中性粒细胞,探讨缺氧对中性粒细胞凋亡的影响。方法：用全反式维甲酸诱导NB-4细胞分化获得中性粒细胞并建立缺氧模型,用流式细胞技术检测缺氧对中性粒细胞凋亡和坏死的影响以及缺氧后CD11b^＋细胞百分率的变化,用免疫荧光技术检测缺氧对中性粒细胞的细胞膜TNFR1表达强度的影响。结果：缺氧后中性粒细胞增殖减缓,凋亡和坏死受到抑制,细胞膜TNFR1表达下降,CD11b^＋细胞百分率稍有增高。结论：缺氧可以减缓中性粒细胞增殖,同时抑制中性粒细胞的凋亡。     

杀菌/通透性增加蛋白的细胞定位

目的 确定人体能产生杀菌/通透性增加蛋白（bactericidal/permeability-increasing protein,BPI)的细胞群体,为研究BPI在细胞内的定位奠定基础。方法 分离人外周血获得嗜中性多形核粒细胞（polymorphonuclear neutrophils,PMN）和单个核细胞,PMN,HL-60及单个核细胞分别进行RNA提取,逆转录PCR反应扩增BPI基因片段和免疫荧光法检测。结果 逆转录PCR在PMN和HL-60细胞中扩增出BPI基因片段,活细胞免疫荧光染色观察到PMN和HL-60能够与抗BPI的单抗所结合而呈阳性。结论 BPI是中性粒系细胞的特异性产物。     

Diffuse panbronchiolitis--a case report

Diffuse panbronchiolitis (DPB) is a chronic progressive obstructive disease of the airways. Of unknown aetiology, it is responsive to macrolide therapy. Although not uncommon among the Japanese, it is rare in other parts of the world. We report a case of DPB in a Chinese patient in Singapore.     

抑制中性粒细胞PD-L1表达对脓毒症T淋巴细胞功能的影响

目的: 探讨脓毒症状态下中性粒细胞对T淋巴细胞功能的影响及相关机制。方法: 采用脂多糖(lipopolysaccharide,LPS)刺激中性粒细胞,模拟脓毒症状态下的体外模型。分离人外周血中性粒细胞并分离出淋巴细胞,分为对照组、LPS组、中性粒细胞组和LPS+中性粒细胞组4个组。采用流式细胞术检测淋巴细胞凋亡、增殖、活性和炎症细胞因子浓度,以及中性粒细胞表面程序性死亡配体1（PD-L1）的表达;流式细胞术检测使用不同浓度PD-L1抗体后的淋巴细胞活性。结果： 与对照组相比,中性粒细胞组和LPS+中性粒细胞组淋巴活性明显降低,凋亡明显增加,细胞增殖明显降低,CD69和CD71表达百分比、平均荧光强度、γ干扰素和IL-2的释放显著降低(P均<0.01);与LPS组相比,LPS+中性粒细胞组能显著抑制淋巴细胞增殖、活化和细胞因子的释放(P均<0.05);与对照组相比,LPS组PD-L1表达明显增加(P<0.05);与对照组相比,中、高浓度组淋巴细胞存活百分比明显增加(P均<0.05),且呈浓度依赖性。 结论： 脓毒症状态下中性粒细胞通过PD-L1/PD-1通路途径抑制T淋巴细胞的功能。     

犬体外循环后中性粒细胞功能变化的研究

目的 研究体外循环后是否存在中性粒细胞的黏附和破坏能力增强,而吞噬杀菌能力减弱的现象.方法 健康成年狼犬12只,随机均分为体外循环(CPB)组和假CPB(Sham)组,CPB组转机100 min后停机;Sham组静置100 min.通过测定CPB后中性粒细胞CD11b/CD18的表达、肺组织髓过氧化物酶(MPO)含量和肺功能等,评估中性粒细胞的黏附功能和对组织的破坏作用;通过测定中性粒细胞吞噬功能、细胞内MPO检测等,评估中性粒细胞的免疫功能.结果 CPB后4 h,CPB组中性粒细胞CD11b、CD18的表达和肺组织MPO活性显著高于肝素化前和Sham组[(55±21)IU/100 g组织vs(30±19)IU/100 g组织,均P<0.01],而PaO2/FiO2显著低于肝素化前和Sham组(319±79 vs 405±101,均P<0.05).细胞的超微结构显示,CPB组中性粒细胞伸出粗大触角.伴随中性粒细胞黏附功能的升高,CPB组中性粒细胞的吞噬功能显著低于体外循环前和对照组(129±53 vs357±92,均P<0.05).但两组动物MPO积分差异无统计学意义.经IL-8刺激后,中性粒细胞MPO释放和氧自由基产生差异无统计学意义(均P>0.05).结论 体外循环能导致中性粒细胞功能失调,包括黏附能力的提高和吞噬功能的下降,这可能是导致体外循环后器官损伤和感染率增加的主要原因.     

Preliminary Study on Drug Susceptibility Profile and Resistance Mechanisms to Macrolides of Clinical Isolates of Non-tuberculous Mycobacteria from China

Objective

Macrolide susceptibility and drug resistance mechanisms of clinical non-tuberculous mycobacteria (NTM) isolates were preliminarily investigated for more accurate diagnosis and treatment of the infection in China.

粒细胞NADPH氧化酶在犬烟雾吸入伤中的变化及意义

本实验利用犬烟雾吸入伤模型。观察了致伤前后粒细胞NADPH氧化酶活性的变化,显示其伤后活性增加,提示粒细胞及其活性氧代谢产物参与了犬烟雾吸入性肺损伤的发生发展,同时观察了血气、病检,肺含水量等指标,证实动物存在严重吸入性肺损伤。     

Factors influencing in vitro production of colony-stimulating factor by mononuclear leukocytes from humans.

The purpose of this study was to identify factors that influence the production of colony-stimulating factor by leukocytes of humans. The use of nonadherent light-density bone marrow cells is semisolid agar cultures to assay the concentrations of colony-stimulating factor in the supernatant of monocyte and mononuclear leukocyte cultures made it possible to distinguish between colony-stimulating factor, which stimulates colony-forming cells directly, and monocyte-dependent stimulating activity, which acts indirectly, by increasing the monocyte production of colony-stimulating factor. Colony-stimulating factor was not detectable in the cytosol of monocytes; that detected in culture must, therefore, have been newly synthesized. Synthesis was enhanced independently by heat-inactivated human serum and by semipurified serum fractions enriched with monocyte-dependent stimulating activity. The kinetics of the production of colony-stimulating factor in the presence and absence of monocyte-dependent stimulating activity indicated that the latter facilitated monocyte production of the former. Factors released from neutrophils were shown to reduce the production of colony-stimulating factor and thr proliferation of colony-forming cells and thus may provide a feedback control mechanism limiting the proliferation of neutrophils.     

溃疡性结肠炎患者外周血中性粒细胞凋亡与细胞因子、粘附分子水平的关系

目的 :观察溃疡性结肠炎患者 PMN凋亡的变化 ,探讨细胞因子对 PMN凋亡的影响。方法 :检测 2 8例 UC病人外周血 PMN凋亡和细胞因子及粘附分子水平。结果 :活动期 UC患者 PMN凋亡率明显低于健康人对照组和缓解期 UC患者。缓解期 UC患者 PMN凋亡率与对照组无差别。不同病情活动期 UC患者 PMN凋亡有显著性差异。活动期 UC患者外周血中 IL - 8、IL- 6、TNF-α、NO、P- sel、ICAM- 1水平均高于对照组和缓解组 ,且与 PMN凋亡呈负相关 ,与病情呈正相关。缓解组患者 IL- 8、IL- 6、TNF- α、NO、P- sel和 ICAM- 1与对照组无显著性差异。结论 :UC患者 PMN凋亡延迟 ,且与病情及疗效密切相关。各种炎性细胞因子产生过多、各种免疫细胞粘附分子表达上调可能是导致 PMN凋亡延迟的重要机制。适度调控 PMN凋亡 ,有可能会成为治疗 UC的有效途径。     

红霉素对急性肺损伤的有限性保护作用

目的探讨红霉素(EM)对急性肺损伤(ALI)动物模型的干预作用。方法经大鼠尾静脉注射无菌油酸(OA)0．15ml／kg复制ALI模型，经大鼠腹腔注射180mg／kg的EM进行预防或治疗，预防组于复制大鼠ALI模型前2h给予EM，治疗组在复制大鼠ALI模型后2、24及48h分别给予EM。分别观测ALI模型大鼠肺组织的病理变化。结果与ALI组相比，EM预防组和EM治疗组大鼠肺组织病理改变显著减轻，其支气管肺泡灌洗液中蛋白含量、中性粒细胞及淋巴细胞均显著减少，但预后无显著差异。结论EM能抑制中性粒细胞和淋巴细胞向损伤肺组织浸润，减轻ALI的肺组织病变，对ALI起有限性保护作用。     

Irreversible sensorineural hearing loss due to clarithromycin

Clarithromycin is a commonly used advanced generation macrolide. This case study reviews a case of an 81 year old woman who developed sensorineural deafness in the right ear after the start of low dose oral clarithromycin for an infective exacerbation of chronic obstructive pulmonary disease. Despite cessation of this drug after only three days, the sensorineural deafness was found to be irreversible. Reversible sensorineural deafness secondary to macrolides has previously been described and evidence in the literature shows that a dose related phenomenon occurs. Research has indicated that transient dysfunction of the outer hair cells could be the possible mechanism. In this case, however, the patient experienced an irreversible sensorineural deafness associated with the start of low dose oral clarithromycin. This is a side effect profile that has not previously been reported.     

雌激素和异黄酮类药物对中性粒细胞粘附分子表达的影响

目的 研究雌激素和异黄酮类药物对中性粒细胞粘附分子表达的影响。方法 利用肿瘤坏死因子（TNFα）处理健康人和缺血性中风患者的中性粒细胞,加用不同学浓度的异黄酮（WZ1、WZ2）和雌激素（WZ3、WZ4）进行干预,采用流式细胞仪定量测定细胞表面的粘附分子表达,结果 （1）10ng/mlTNFα对健康人中性粒细胞有明显的激活作用。可使CD18表达上调10%,CD62L平均荧光强度下降15%,阳性细胞百分数下降30%。（2）异黄酮类药物WZ1、WZ2对中性粒细胞表面的CD18和CD62L表达基本上没有影响。（3）用雌激素类药物WZ3、WZ4预处理中性粒细胞后再加TNFα激活,可以使中性粒细胞表面粘附分子CD18的平均荧光强度下降8%,CD62L的平均荧光强度增加15%,CD62L的阳性细胞百分率增加20%;如果先用TNFα激活中性粒细胞然后再用WZ3、WZ4处理,中性粒细胞粘附分子的表达与单纯TNFα组之间无显著性差别。（4）TNFα处理缺血性中风患者中性粒细胞处理,中性粒细胞粘附分子的表达与单纯TNFα组之间无显著性差别。（4）TNFα处理缺血性中风患者中性粒细胞后,CD18表达增加20%,CD62L的平均荧光强度和阳性细胞百分数均下降30%,与健康对照组相比有统计学差异,雌激素预处理后可以抑制中性粒细胞的激活。结论（１）ＴＮＦα是中性粒细胞强有力的激活剂,在中风的发生、发展过程发挥了重要作用。（２）异黄酮类药物对中性粒细胞在细胞因子刺激下粘附分子的表达没有很大影响,提示该类药物对心脑血管的保护作用可能不通过此途径发挥作用。（３）雌激素类药物预处理中性粒细胞,可以抑制ＴＮＦα对它们的激活作用。降低粘附分子的表达,减少粘附;雌激素类药物可以用来预防中性粒细胞与内皮细胞的粘附,从而可降低缺血性中风的发生;而雌激素类药物对于已经被ＴＮＦα激活的中性粒细胞粘附分子的表达没有影响,提示该类药物对于抗粘附缺乏治疗意义。     

雌激素和异黄酮类药物对中性粒细胞粘附分子表达的影响

目的　研究雌激素和异黄酮类药物对中性粒细胞粘附分子表达的影响。方法　利用肿瘤坏死因子 ( TNFα)处理健康人和缺血性中风患者的中性粒细胞 ,加用不同浓度的异黄酮 ( WZ1 、WZ2 )和雌激素 ( WZ3 、WZ4)进行干预 ,采用流式细胞仪定量测定细胞表面的粘附分子表达。结果　 110 ng/ml TNFα对健康人中性粒细胞有明显的激活作用 ,可使 CD18表达上调 10 % ,CD6 2 L平均荧光强度下降 15 % ,阳性细胞百分数下降 30 %。 2异黄酮类药物 WZ1 、WZ2 对中性粒细胞表面的 CD18和 CD6 2 L表达基本上没有影响。 3用雌激素类药物 WZ3 、WZ4预处理中性粒细胞后再加 TNFα激活 ,可以使中性粒细胞表面粘附分子 CD18的平均荧光强度下降 8% ,CD6 2 L的平均荧光强度增加 15 % ,CD6 2 L 的阳性细胞百分率增加 2 0 % ;如果先用 TNFα激活中性粒细胞然后再用 WZ3 、WZ4处理 ,中性粒细胞粘附分子的表达与单纯 TNFα组之间无显著性差别。 4TNFα处理缺血性中风患者中性粒细胞后 ,CD18表达增加 2 0 % ,CD6 2 L 的平均荧光强度和阳性细胞百分数均下降 30 % ,与健康对照组相比有统计学差异 ;雌激素预处理后可以抑制中性粒细胞的激活。结论　 1TNFα是中性粒细胞强有力的激活剂 ,在中风的发生、发展过程发挥了重要作用。 2异黄酮类药     

Azithromycin inhibits neutrophil accumulation in airways by affecting interleukin-17 downstream signals

Background  Azithromycin can reduce neutrophil accumulation in neutrophilic pulmonary diseases. However, the precise mechanism behind this action remains unknown. Our experiment assessed whether azithromycin inhibits neutrophil accumulation in the airways by affecting interleukin-17 (IL-17) downstream signals.

Methods  Mice were pretreated with azithromycin before murine IL-17A (mIL-17) stimulation. After the mIL-17 stimulation, the levels of six neutrophil-mobilizing cytokines were determined by enzyme-linked immunosorbent assay (ELISA) tests in bronchoalveolar lavage (BAL) fluid; IL-6, CXC chemokine ligand-1 (CXCL-1), CXCL-5, macrophage inflammatory protein-2 (MIP-2), granulocyte colony-stimulating factor (G-CSF), and granulocyte macrophage colony-stimulating factor (GM-CSF). The number of neutrophils in BAL fluid were evaluated by cytospin preparations.

Results  (1) Azithromycin pretreatment significantly inhibited both the release of three neutrophil-mobilizing cytokines (MIP-2, CXCL-5 and GM-CSF) and the accumulation of neutrophils in airways caused by mIL-17 stimulation. (2) The levels of three neutrophil-mobilizing cytokines (IL-6, MIP-2 and GM-CSF) were positively correlated with the numbers of neutrophil in BAL fluid.

Conclusions  Azithromycin can inhibit neutrophil accumulation in the airways by affecting IL-17 downstream signals. This finding suggests that macrolide antibiotic application might be useful in prevention of neutrophilic pulmonary diseases characterized by high levels of IL-17.

     

重症肺炎患者中性粒细胞胞吐作用的改变

目的　研究重症肺炎患者中性粒细胞的功能改变。方法　采用免疫化学法测定 32例重症肺炎患者中性粒细胞胞吐作用和氧化酶反应的变化 ,并与健康对照组比较。结果　重症肺炎患者血液中性粒细胞的基础和PMA刺激的胞吐作用均明显下降 (P <0 .0 1) ;重症肺炎患者的基础和PMA刺激的反应性氧属 (ROS)产生明显增加 ;重症肺炎患者肺部中性粒细胞释放乳铁蛋白、髓过氧化物酶 (MPO)和ROS增加 ,PMA刺激后相似 ;肺炎的严重性与血液中性粒细胞释放乳铁蛋白呈负相关。结论　重症肺炎患者中性粒细胞胞吐作用明显受损 ,与预后相关。     

慢性支气管炎与肺气肿大鼠气道炎症的实验研究

目的　探讨慢性支气管炎 (慢支炎 )与肺气肿气道炎症的特点及红霉素对气道炎症的干预作用。方法　将 43只雄性Wistar大鼠随机分为正常组 ,对照组 ,慢支炎组 ,慢支炎并肺气肿组 ,低、高剂量红霉素治疗组及低、高剂量红霉素预防组共 8组 ,应用气管内注入脂多糖和烟熏的方法复制慢支炎、慢支炎并肺气肿动物模型 ,收集支气管肺泡灌洗液 (BALF)进行细胞学计数和分类检查 ,并对大鼠肺脏进行病理学检查。结果　慢支炎组BALF中有核细胞数及中性粒细胞分类计数较对照组显著增高 (P <0 0 5) ;慢支炎并肺气肿组BALF中有核细胞数、中性粒细胞分类计数及单核巨噬细胞绝对数均显著增加 ,肺泡间隔破坏 ,肺气肿形成。慢支炎组、慢支炎并肺气肿组与正常组比较病理总分均显著增高 (P <0 0 1 ) ,慢支炎并肺气肿组病理积分较慢支炎组增高。红霉素治疗组及预防组BALF中中性粒细胞数显著降低 ,病理总分也有不同程度降低。结论　多种炎性细胞 ,尤其是中性粒细胞、肺泡巨噬细胞在慢支炎和肺气肿气道炎症中起重要作用 ,随病情加重 ,肺泡巨噬细胞浸润更加明显 ;红霉素对慢支炎与肺气肿气道炎症具有一定的防治作用 ,其疗效与剂量和疗程有关 ,其机理与抑制慢性阻塞性肺疾病气道炎症中性粒细胞聚集和活化有关     

氧自由基与肺损伤——Ⅲ.钨对实验性肺损伤的影响

钨酸钠对油酸(OA)所致的大鼠肺损伤有明显保护作用,表现在肺系数,支气管肺泡灌洗液中嗜中性白细胞(PMN)和丙二醛、肺泡内RBC与PMN渗出比对照组显著降低;注入OA后5、15min的外周血中白细胞总数与PMN亦无明显下降。其机制可能与钨能抑制黄嘌呤氧化酶,使氧自由基产生减少有关。