弥漫性泛细支气管炎气道黏液高分泌的研究进展

弥漫性泛细支气管炎是一种小气道慢性炎症性疾病，气道黏液高分泌即痰液增多是该病主要临床表现之一。其黏液高分泌的机制至今尚不清楚，相关研究表明与绿脓杆菌、中性粒细胞、炎症介质及黏蛋白基因的多态性有关。另外，大环内酯类抗生素对弥漫性泛细支气管炎的气道高分泌有显著治疗作用。     

表皮生长因子受体与气道黏液高分泌

黏液高分泌是慢性气道炎症性疾病的重要病理特征之一,也是患者病情加剧恶化的重要因素.表皮生长因子受体(EGFR)是一种受体酪氨酸激酶(RTK),在气道黏液高分泌的信号转导通路中居于中心地位,各种炎性介质、病原微生物及其产物、活性氧等可通过多种信号转导通路活化EGFR,活化相关转录因子引起黏蛋白5AC基因表达上调.明确EGFR上下游信号通路势必对慢性气道炎症性疾病的防治提供指导.     

Human epidermal growth factor receptor residue covalently cross-linked to epidermal growth factor.

An epidermal growth factor (EGF) receptor monoclonal antibody (mAb), mAb LA22, was used to analyze the covalent coupling of human EGF receptors to mouse EGF by the amine-reactive cross-linking agent disuccinimidyl suberate. A soluble Mr 105,000 truncated form of the receptor secreted by A-431 epidermoid carcinoma cells and consisting of the ligand-binding extracellular domain was cross-linked to 125I-labeled EGF. Digestion of this complex with an endoproteinase that specifically cleaves at the COOH side of glutamyl residue released a single radiolabeled glycosylated fragment of Mr 18,000 that reacted with mAb LA22. As the epitope for mAb LA22 resided between Ala-351 and Asp-364 of the mature receptor, this result localized the cross-linked receptor residue(s) to the 47-amino acid interval from Phe-321 to Glu-367. The receptor residue(s) involved in the covalent coupling of rat 125I-labeled transforming growth factor alpha was similarly localized to this region of the receptor. This receptor interval, which included two glycosylated asparaginyl residues at positions 328 and 337, contained but three amino acid residues that were potentially reactive with disuccinimidyl suberate: Lys-332, Lys-333, and Lys-336. Characterization of mAb LA22-reactive 125I-EGF-labeled receptor fragments generated by an endoproteinase specific for the COOH side of lysyl residue placed the NH2 termini of the two smallest fragments between the glycosylated residues Asn-328 and Asn-337. These results indicated that disuccinimidyl suberate cross-linked the NH2 group of EGF residue Asn-1 to the human EGF receptor residue Lys-336. Our results further suggest that EGF and transforming growth factor alpha, two members of the EGF family of peptide growth factors, interact with closely apposed or identical features of the receptor.     

致敏大鼠气道黏液异常分泌与生长因子受体信号调节

目的探讨致敏大鼠黏液高分泌与转化生长因子α(transfroming growth factor-α,TGF-α)、表皮生长因子受体(epidermal growth factor receptor,EGFR)通路之间的关系。方法应用析因设计方法,随机将大鼠分为12组,每组4只。采用卵蛋白(OVA)腹腔注射加雾化吸入致敏;HE染色和AB-PAS染色测定杯状细胞化生/增生(GCH/GCM)指数;免疫组织化学检测支气管上皮细胞MUC5AC蛋白和EGFR蛋白表达水平;放射免疫方法检测血清中TGF-α。结果①哮喘气道上皮杯状细胞增生肥大,大量黏液形成,小气道内有黏液栓形成。MUC5AC蛋白表达和GCH指数明显增高,呈动态演变。且与EGFR蛋白表达和血清TGF-α水平呈正相关。②析因分析表明,不同处理因素,MUC5AC蛋白表达、GCH/GCM指数、EGFR蛋白表达和血清TGF-α水平差异有统计学意义(P<0.01),MUC5AC除外。不同的处理时间,MUC5AC蛋白表达、GCH/GCM指数、EGFR蛋白表达和血清TGF-α水平差异有统计学意义(P<0.01)。处理因素和处理时间有交互作用(P>0.05)。结论哮喘组MUC5AC蛋白表达和GCH/GCM指数明显增高,且随着激发时间呈动态演变。Genistein通过抑制EGFR通路活性降低MUC5AC高表达和GCH/GCM指数。     

茶黄素和表皮生长因子受体对气道黏液分泌的影响

目的 探讨茶黄素对炎症反应时气道上皮细胞黏液分泌的影响.方法 通过人中性粒细胞弹性蛋白酶(HNE)刺激人肺腺癌细胞A549,构建炎症反应时气道黏液高分泌模型,以茶黄素和表皮生长因子受体(EGFR)阻断剂表皮生长因子受体阻断剂(AG1478)进行干预,观察黏蛋白5AC(MUCSAC)、EGFR、磷酸化EGFR(P-EGFR)、磷酸化细胞外信号调节激酶1/2(P-ERK1/2)、兔抗磷酸化p38(P-p38)及磷酸化JNK丝裂原活化蛋白激酶(P-JNK)的表达.用四甲基偶氮唑盐法测定细胞活性,再将A549细胞分为对照组、HNE处理组、茶黄素组、AG1478组和茶黄素+AG1478组.用逆转录PCR方法检测各组MUC5AC mRNA、EGFR mRNA的变化;Western blot法检测EGFR、P-EGFR、P-ERK1/2、P-p38和P-JNK蛋白的表达;酶联免疫吸附测定法观察MUCSAC蛋白表达的变化,并用细胞免疫激光共聚焦显微镜观察作用前后黏蛋白的分布.两样本均数间比较采用t检验,多样本均数间比较采用单因素方差分析.结果 HNE处理组MUC5AC的mRNA和蛋白的积分吸光度值分别为0.99±0.03和(169±6)μg/mg,EGFR mRNA和蛋白表达的积分吸光度值分别为0.98±0.02和(0.89±0.03)μg/mg,均较对照组[0.53±0.02、(105±4)μg/mg和0.61±0.11、0.21±0.05]明显升高;P-EGFR、P-ERK1/2的蛋白表达也较对照组显著增加,而P-p38的表达则有较低幅度的增强,P-JNK无明显变化.给予茶黄素及AG1478预处理后,与HNE刺激组相比,EGFR、P-EGFR、P-ERK1/2、P-p38均明显下调,P-JNK无相应改变;而茶黄素+AG1478组MUCSAC mRNA和MUC5AC的下调较单独用茶黄素或AG1478处理更为明显,其积分吸光度值分别为0.20±0.02和(125±3)μg/mg,差异均有统计学意义(t值分别为3.02和1405.94,均P<0.05).结论 茶黄素可通过下调EGFR水平、减少EGFR的活化、部分阻遏EGFR信号转导途径及细胞外信号调节激酶来实现对下游途径的影响,从而发挥茶黄素抑制炎性气道黏液高分泌形成的作用.     

Therapeutic approaches to mucus hypersecretion

Mucolytic and related agents have been in use since prehistoric times. Although widely prescribed and used extensively in over-the-counter preparations, their efficacy and mechanisms of action remain in doubt. These agents belong to several distinct chemical classes. Mucolytic agents such as N-acetyl-cysteine are thiols with a free-sulfhydryl group. They are assumed to break disulfide bonds between gelforming mucins and thus reduce mucus viscosity. Mucokinetic agents are thiols with a blocked sulfhydryl group. Expectorants such as guaifenesin increase mucus secretion. They may act as irritants to gastric vagal receptors, and recruit efferent parasympathetic reflexes that cause glandular exocytosis of a less viscous mucus mixture. Cough may be provoked. This combination may flush tenacious, congealed mucopurulent material from obstructed small airways and lead to a temporary improvement in dyspnea or the work of breathing. The roles of anticholinergic agents, DNase, and other drugs are also discussed with regard to their roles in reducing mucus production in rhinitis and other airway diseases.     

Human epidermal growth factor receptor 2 expression in mixed gastric carcinoma

AIM: To investigate human epidermal growth factor receptor 2 (HER2) amplification and protein expression in mixed gastric carcinoma.METHODS: Fluorescence in situ hybridization and immunohistochemistry were used to detect HER2 amplification and protein expression in 277 cases of mixed gastric carcinoma. Protein staining intensity was rate as 1+, 2+, or 3+.RESULTS: Of the 277 cases, 114 (41.2%) expressed HER2 protein. HER2 3+ staining was observed in 28/277 (10.1%) cases, 2+ in 37/277 (13.4%) cases, and 1+ in 49/277 (17.7%) cases. A HER2 amplification rate of 17% was detected, of which 25/28 (89.3%) were observed in the HER2 3+ staining group, 17/37 (45.9%) in 2+, and 5/49 (10.2%) in 1+. Of the 47 patients with HER2 amplification who received chemotherapy plus trastuzumab, 22 demonstrated median progression-free and overall survivals of 9.1 mo and 16.7 mo, respectively, which were significantly better than those achieved with chemotherapy alone (5.6 mo and 12.1 mo, respectively) in 19 previously treated patients (Ps < 0.05).CONCLUSION: HER2 detection in mixed gastric carcinoma displays high heterogeneity. Relatively quantitative parameters are needed for assessing the level of HER2 amplification and protein expression.     

表皮生长因子及其受体在糜烂胃黏膜中的表达

目的:观察胃窦黏膜糜烂区与糜烂旁胃黏膜、慢性萎缩性胃炎中表皮生长因子(epidermal growth factor,EGF)及其受体(epidermal growth factor receptor,EGFR)的表达,探讨其在胃黏膜损伤修复中的意义.方法:选择经胃镜及病理确诊的慢性萎缩性胃炎伴胃窦黏膜糜烂患者50例,距糜烂区3cm处40例,无糜烂慢性萎缩性胃炎40例,采用免疫组织化学染色法测EGF及EGFR的表达.结果:胃窦黏膜糜烂区EGF、EGFR阳性表达率分别为40%和30%,明显高于糜烂旁胃黏膜15%和10%及无糜烂慢性萎缩性胃炎20%和12.5%的阳性表达率(P<0.05),有统计学意义,无糜烂慢性萎缩性胃炎组略高于糜烂旁胃黏膜组,但无统计学差异.结论:EGF、EGFR在胃黏膜损伤后高表达,对促进胃黏膜修复有着重要意义.     

EGFR在结直肠癌组织中的表达及其意义

目的:探讨结直肠癌患者肿瘤组织中表皮生长因子受体(epidermal growth factor,EGFR)过表达与临床病理特征及其预后的关系和意义.方法:收集2004-01/2010-04在中国人民解放军南京军区福州总医院手术切除的结直肠癌组织标本1 228例,选择其中资料完整的病例1 100例,采用EliVisio...     

Decreased expression of hepatic epidermal growth factor receptor gene in diabetic mice

The expression of hepatic epidermal growth factor (EGF) receptor gene was studied in genetically diabetic (C57BL/KsJ db/db) mice and streptozotocin-induced diabetic mice. The binding of 125I-labelled EGF to hepatic membrane preparations of genetically diabetic mice was only 35% of that of non-diabetic mice. Levels of EGF receptor messenger RNAs (10 and 6 kb) in the liver of the diabetic animals were also reduced by about 75%. In streptozotocin-induced diabetic mice, levels of hepatic EGF binding and messenger RNAs for EGF receptor were decreased to 27 and 30% of control levels respectively, at 5 weeks after injection of the drug. There were, however, no significant differences in levels of messenger RNAs for the structural protein beta-actin in the liver. In addition, levels of EGF receptor messenger RNAs in the kidney were similar between control and the two kinds of diabetic mice. Daily administration of insulin to the streptozotocin-induced diabetic mice increased the hepatic levels of EGF receptor messenger RNAs to almost normal levels. These results indicate that EGF binding to its receptor decreases in the liver of diabetic mice, involving alterations in the level of EGF receptor messenger RNAs, and that insulin is important for the regulation of EGF receptor gene expression in the liver but not in the kidney.     

Reduced expression of epidermal growth factor receptor related protein in gastric cancer

OBJECTIVES: The recently cloned epidermal growth factor receptor related protein (ERRP) has been proposed to be a negative regulator of the epidermal growth factor receptor (EGFR). Because of the causal involvement of EGFR and its ligands in gastric cancer growth, we investigated expression of ERRP and cell proliferation in human gastric cancer. METHODS: We examined ERRP expression and localisation in surgical specimens of gastric cancers from 47 patients versus non-malignant gastric mucosa and determined their relationship to cell proliferation and differentiation. We also examined expression of ERRP by western blotting in three different gastric cancer cell lines. To further determine the functional properties of ERRP, we examined the effect of ERRP on epidermal growth factor (EGF) induced EGFR phosphorylation essential for its activation in MKN-28 gastric cancer cells. RESULTS: ERRP expression was dramatically reduced in gastric cancers (34% of all specimens positive) compared with non-malignant gastric mucosa (66% of specimens positive). Expression of ERRP in cancer cells inversely correlated with cell proliferation and grade of malignancy. Cell lines derived from metastatic gastric cancers had reduced ERRP expression compared with cell lines derived from a non-metastatic cancer. Exogenous ERRP protein markedly inhibited EGF induced EGFR phosphorylation in gastric cancer cells providing a novel molecular mechanism of its action. CONCLUSIONS: Our data indicate that downregulation of ERRP could play an important role in gastric cancer differentiation and progression. ERRP is a negative regulator of tumour cell proliferation and may exert its inhibitory effect, in part, by attenuating EGFR activation.     

Genetic predisposition to diffuse panbronchiolitis

Diffuse panbronchiolitis is characterized by chronic inflammation in respiratory bronchioles and sinobronchial infection. The pathophysiology accompanying the persistent bacterial infection is noteworthy for the accumulation of lymphocytes and foamy macrophages around the small airways, for mucus hypersecretion, and for the number of neutrophils in the large airways. Until the establishment of long-term macrolide therapy, the prognosis was generally poor. Case studies of diffuse panbronchiolitis in East Asians, including Japanese, Koreans and Chinese, have frequently been reported, and genetic predisposition to the disease has been assumed in Asians. Immunogenetic studies revealed a strong association with human leukocyte antigen (HLA)-B54 in Japanese, whereas an association with HLA-A11 was reported in Koreans. These findings imply that a major susceptibility gene may be located between the HLA-A and HLA-B loci on the short arm of human chromosome 6. We have recently cloned novel mucin-like genes in this candidate region. In addition to accumulated knowledge of classical HLA genes and mucin genes, further analysis of newly identified genes may provide insights into the pathogenesis of the disease.     

肝内胆管癌形成过程中表皮生长因子及其受体与转化生长因子-α的表达意义

通过免疫组织化学的方法，动态观察表皮生长因子(EGF)及其受体(EGFR)和转化生长因子-α(TGF-α)在叙利亚地鼠肝内胆管癌(ICC)形成过程中的表达情况，探讨上述因子在ICC形成过程中的作用。     

The epidermal growth factor receptor family

The epidermal growth factor receptor family consists of four receptor genes and at least 11 ligands, several of which are produced in different protein forms. They create an interacting system that has the ability to receive and process information that results in multiple outputs. The family has an important role in directing and coordinating many normal processes, including growth and development, normal tissue turnover and wound healing. Its members are also aberrantly activated by overexpression or mutation in many common human tumour types and as such have been the target for anticancer drug development.     

Correlation of human epidermal growth factor receptor protein expression and colorectal cancer

AIM: To investigate the correlation between human epidermal growth factor receptor (HER-2) protein expression and colorectal cancer (CRC) using a case-control study and meta-analysis.METHODS: Tumor tissue specimens from 162 CRC patients were selected for the case group. Fifty cases were randomly selected, and normal CRC tissue at least 10 cm away from the tumor margins of these cases was used to generate the control group. The expression of the HER-2 protein in the 162 CRC tissue samples and the 50 adjacent normal mucosa tissue samples was detected via immunohistochemistry. The experimental data were analyzed using SPSS 18.0 software, and R software version 3.1.0 was utilized for further verification.RESULTS: The expression of HER-2 protein in the 162 CRC tissue samples was significantly higher than in the normal tissue specimens. The data showed that the expression of HER-2 in CRC was related to the Dukes’ stage, the depth of invasion and lymph node metastasis. The HER-2-positive patients had lower 3- and 5-year OS rates than the HER-2-negative patients, but there was no significant difference. However, there was a statistically significant difference in the 3- and 5-year disease-free survival (DFS) rates of HER-2-positive and HER-2-negative patients. The results of the meta-analysis showed that the expression of HER-2 in CRC patients was statistically significantly increased over that of healthy people. The 3-year DFS rate in HER-2-positive patients was markedly lower than that in HER-2-negative patients.CONCLUSION: Down-regulation of HER-2 expression might be a dependable strategy for CRC therapy.     

Clinical efficacy of the FLUTTER device for airway mucus clearance in patients with diffuse panbronchiolitis

Abstract Expectoration of mucus is important in preventing the development of airway inflammation in patients with diffuse panbronchiolitis (DPB). To evaluate the clinical efficacy of the FLUTTER device in clearing mucus from the airways of patients with DPB who have difficulty expectorating, we assessed pulmonary function and symptoms in patients treated with FLUTTER. Eight patients in a stable clinical condition with DPB were included in the study. The study was divided into two consecutive, 1-week periods. The initial week was an observation week. During the following week, patients used FLUTTER four times daily. Expectorated sputum was collected in a container and weighed every day during 2 weeks. Pulmonary function, partial oxygen pressure and partial carbon dioxide pressure in arterial blood were measured in all patients on the last day of the observation week and the FLUTTER treatment week. A symptom score for difficulty of expectoration was determined by questionnaire. A pneumothorax developed in one patient during using FLUTTER. The mean daily sputum weight and peak expiratory flow rate increased significantly after treatment with FLUTTER ( P < 0.04 and P< 0.02, respectively). Symptom score improved significantly after using FLUTTER ( P < 0.02). We conclude that the use of FLUTTER is effective in clearing mucus from the airways. However, the development of a pneumothorax may complicate use of the procedure in some cases.     

过氧化物酶体增殖物激活受体γ及其配体与气道黏液高分泌

气道黏液高分泌足慢性气道疾病如慢性阻塞性肺疾病、支气管哮喘、肺囊性纤维化、支气管扩张等共同而重要的病理特征之一,与气道疾病的发生、发展转归有密切关系.过氧化物酶体增殖物激活受体γ是一类由致密分子构成依赖配体激活的核转录因子.近年研究认为,过氧化物酶体增殖物激活受体γ及其配体能拮抗气道炎症反应,能对气道黏液分泌发挥调节作用.     

过氧化物酶体增殖物激活受体γ及其配体与气道黏液高分泌

气道黏液高分泌足慢性气道疾病如慢性阻塞性肺疾病、支气管哮喘、肺囊性纤维化、支气管扩张等共同而重要的病理特征之一,与气道疾病的发生、发展转归有密切关系.过氧化物酶体增殖物激活受体γ是一类由致密分子构成依赖配体激活的核转录因子.近年研究认为,过氧化物酶体增殖物激活受体γ及其配体能拮抗气道炎症反应,能对气道黏液分泌发挥调节作用.