目的 探讨负性调节细胞CD4⁺CD25⁺T及其相关细胞因子在慢性阻塞性肺病(COPD)患者外周血中的表达与合并细菌感染的相关性。方法 纳入2018年1月—2019年12月间收治的66例COPD患者作为研究对象,其中急性加重期COPD患者(AECOPD)36例、稳定期患者30例,并纳入同期体检健康者30例作为对照组。对所有纳入的研究对象外周血标本中的CD4⁺CD25⁺T调节性T细胞及其相关细胞因子[白介素-4(IL-4)、白介素-10(IL-10)、干扰素-γ(IFN-γ)]表达水平进行检测,分析相关指标水平与COPD是否合并细菌感染的关系,及预测细菌感染的效能。结果 AECOPD和稳定期COPD患者CD4⁺、CD4⁺CD25⁺、IFN-γ/IL-4水平均低于对照组(P<0.05),IL-4、IL-10水均高于对照组(P<0.05);AECOPD患者IFN-γ水平高于对照组(P<0.05);AECOPD患者CD4⁺、CD4⁺CD25⁺水平低于稳定期COPD患者(P<0.05),IL-4、IL-10、IFN-γ均高于稳定期COPD患者(P<0.05);CD4⁺、CD4⁺CD25⁺水平与IL4、IFN-γ均呈负相关关系(P<0.05),CD4⁺水平与IL-10呈负相关关系(P<0.05);COPD合并感染者CD4⁺水平低于未合并感染者(P<0.05),IL-4、IFN-γ水平均高于未合并感染者(P<0.05);COPD合并革兰氏阴性菌感染者CD4⁺CD25⁺水平低于未合并感染者(P<0.05),IL-10水平均高于未合并感染者(P<0.05);CD4⁺、IL-4、IL-10、IFN-γ均是预测COPD患者合并细菌感染的有效指标(P<0.05),其中IL-4和IFN-γ效能较高。结论 CD4⁺、CD4⁺CD25⁺Treg细胞及其相关细胞因子参与COPD发生发展和患者细菌感染,监测其水平变化有利于为临床诊治提供信息。

Objective To investigate the correlation between the expressions of negative regulatory cell CD4⁺CD25⁺T and its related cytokines in peripheral blood and bacterial infection of patients with chronic obstructive pulmonary disease (COPD). Methods Sixty-six COPD patients admitted between January 2018 and December 2019 were included as the research subjects, including 36 patients with acute exacerbation of COPD (AECOPD) and 30 patients with stable COPD. Another 30 healthy people undergoing physical examination during the same period were included in control group. The expression levels of CD4⁺CD25⁺ regulatory T cell and its related cytokines [interleukin-4 (IL-4), interleukin-10 (IL-10), interferon-γ (IFN-γ)] in the peripheral blood samples were detected among the included subjects. The relationship between levels of related indicators and presence or absence of bacterial infection in COPD and the efficacy of predicting infection were analyzed. Results The levels of CD4⁺, CD4⁺CD25⁺ and IFN-γ/IL-4 in patients with AECOPD and patients with stable COPD were lower than those in control group (P<0.05), while the levels of IL-4 and IL-10 were higher than those in control group (P<0.05). The IFN-γ level of AECOPD patients was higher than that of control group (P<0.05). The levels of CD4⁺ and CD4⁺CD25⁺of AECOPD patients were lower than those of stable COPD patients (P<0.05), while the levels of IL-4, IL-10 and IFN-γ were all higher than those of stable COPD patients (P<0.05). The levels of CD4⁺ and CD4⁺CD25⁺were negatively correlated with IL-4 and IFN-γ (P<0.05), and the CD4⁺level was negatively correlated with IL-10 (P<0.05). The CD4⁺ level in COPD patients with infection was lower than that in patients without infection (P<0.05), while the levels of IL-4 and IFN-γ were higher than those in patients without infection (P<0.05). The CD4⁺CD25⁺level of COPD patients with Gram-negative bacteria infection was lower than that of patients without infection (P<0.05), while the IL-10 level was higher than that of patients without infection (P<0.05). CD4⁺, IL-4, IL-10 and IFN-γ were effective indicators in predicting bacterial infection in COPD patients (P<0.05), and IL-4 and IFN-γ had higher efficacy. Conclusions CD4⁺, CD4⁺CD25⁺ T cell and related cytokines are involved in the occurrence and development of COPD and bacterial infection in patients. Monitoring changes of those levels is helpful to provide information for clinical diagnosis and treatment.

目的 分析ABCC2基因表达水平与肺腺癌预后之间的关联性,并对其影响机制进行初步探索。 方法 采用TCGA数据库和HPA数据库对肺腺癌病人癌组织和癌旁组织基因表达数据进行差异性分析,单因素及多因素COX回归评估ABCC2与肺腺癌预后之间的关联性,GSEA用于探讨与ABCC2显著关联的信号通路。 结果 ABCC2在肺腺癌肿瘤组织中存在过表达现象,Kaplan-Meier生存分析曲线结果显示ABCC2基因过表达使肺腺癌病人的死亡风险显著升高(HR=1.46,95%CI=1.09~1.95; P=0.010)。单因素及多因素COX回归结果显示ABCC2基因过表达是肺腺癌病人不良预后的独立危险因素。GSEA结果显示ABCC2可能通过调节药物代谢从而对肺腺癌的发展进行调控。 结论 ABCC2基因过表达使肺腺癌病人的死亡风险显著升高,ABCC2可能是肺腺癌不良预后的潜在分子生物标志物。

Objective To estimate the association between ABCC2 mRNA expression and the prognosis of lung adenocarcinoma and explore the potential influencing mechanism.Methods Difference analysis was used to evaluate the gene expression in tumor tissues and adjacent normal tissues based on The Cancer Genome Atlas database and Human Protein Atlas database.Multivariate COX regression and Kaplan-Meier analysis were performed to evaluate the association between ABCC2 gene expression and the prognosis of lung adenocarcinoma.Gene-set enrichment analysis (GSEA) was performed to screen differentially enriched pathways associated with the ABCC2 high expression phenotype.Results ABCC2 was overexpressed in lung adenocarcinoma tumor tissues compared with adjacent normal tissues.Kaplan-Meier survival analysis showed a significant relationship between ABCC2 mRNA expression and lung adenocarcinoma prognosis (HR=1.16,95% CI=1.09-1.95; P=0.010).Univariate and multivariate Cox regression analysis showed that ABCC2 mRNA expression was an independent risk factor affecting the survival of patients with lung adenocarcinoma.The results of GSEA suggested that ABCC2 may influence the development of lung adenocarcinoma by regulating the metabolism of targeted drug the treatment.Conclusions ABCC2 overexpression can significantly increase the risk of death in patients with lung adenocarcinoma,ABCC2 may be a potential molecular marker for poor prognosis in lung adenocarcinoma.

目的 观察紧密连接蛋白在高尿酸血症致大鼠肾损害模型中的表达变化以及非布司他的干预疗效。方法 将SD大鼠分为正常组,高尿酸血症组(模型组),非布司他组(干预组);氧嗪酸联合尿酸诱导制作高尿酸血症大鼠模型,给予非布司他进行干预,分别于6周后检测各组大鼠血中尿素氮(BUN)、血肌酐(Scr)、尿酸(UA)水平,免疫组化及RT-PCR方法检测紧密连接蛋白包括膜周蛋白-1(ZO-1)、跨膜蛋白(occludin) 的表达变化,采用Masson染色检测大鼠肾间质病理改变。结果 6周时,模型组、干预组ZO-1、occludin表达较正常组降低(均P<0.05);干预组ZO-1、occludin表达较模型组增加,差异有统计学意义(均P<0.05),与正常组相比,模型组、干预组RIF指数均增高(均P<0.05),干预组RIF指数低于模型组,高于正常组(均P<0.05)。结论 紧密连接蛋白表达的降低在高尿酸血症肾间质纤维化发展过程中起着举足轻重的作用,并与血尿酸水平及肾功能损害密切相关。非布司他通过降低血尿酸水平,能改善紧密连接蛋白的表达,延缓肾功能损害,起到肾保护作用。

Objective To observe the expression of tight junction protein in hyperuricemia induced renal damage model in rats and the intervention effect of febuxostat. Methods SD rats were randomly divided into three groups: normal control group, model control group, febuxostat treatment group. Hyperuricemia was induced in rats with oxonic acid per time for three times per day, by gavage and combined with uric acid added in drinking water, while febuxostat were administered by gavage in febuxostat treatment group.The blood of rats were collected to analyse the differences of control, model and treatment group on changes of blood urea nitrogen (BUN), creatinine (Cr), uric acid (UA). Immunohistochemistry was used to assay ZO-1 and occludin protein expression and quantitive real time PCR to detect the expression of ZO-1 and occludin in renal tissue of renal interstitial fibrosis model rats induced by hyperuricemia. Paraffin section of kidney was maked and then performed Masson staining to make sure the model is successful. Results At 6 weeks, the expressions of ZO-1 and occludin in the model group and treatment group were lower than those in the normal group (all P<0.05). The expressions of ZO-1 and occludin in the treatment group were higher than those in the model group (all P<0.05). Compared with the normal group, the RIF index in the model group and treatment group were higher (all P<0.05), and the RIF index in the treatment group was lower than that in the model group and higher than that in the normal group (all P<0.05). Conclusion The downregulated expression of ZO-1 and occludin plays a crucial role during the development of hyperuricemia in renal interstitial fibrosis, and are closely related to UA level and renal function impairment. Febuxostat may improve the expression of tight junction by downregurating UA, reduce renal fuction impairment and play a role in renal protection.

目的 探讨SNCG蛋白在卵巢癌组织中的表达情况及临床意义,明确SNCG在人卵巢癌中的表达情况及其恶性程度的关系,为临床卵巢癌的诊断、治疗及预后提供理论依据。方法 收集2010年1月—2015年1月石河子大学医学院第一附属医院收治的具有完整临床病理资料和石蜡切片的119例卵巢癌以及50例正常卵巢患者,用免疫组化方法检测组织中SNCG的表达情况,并分析SNCG的表达与卵巢癌患者临床病理特征及预后的关系。结果 SNCG在卵巢癌组织中的表达高于正常卵巢组织(χ²=73.575,P＜0.001);SNCG的表达与卵巢癌患者的肿瘤分期、分化程度、淋巴结转移、达到满意减瘤术、CA125以及HE4水平相关,差异均有统计学意义(P＜0.05);与卵巢癌肿瘤的原发部位、腹水、复发及化疗耐药无关,差异无统计学意义(P＞0.05);SNCG的过表达与HGSOC患者的PFS、OS相关,差异有统计学意义(P＜0.05);多变量Cox比例风险模型分析显示SNCG是HGSOC患者的独立预后因素,与PFS(HR=2.107,95%CI:1.014～3.795,P=0.034)、OS(HR=1.238,95%CI:0.716～1.928,P=0.047)相关。结论 SNCG在卵巢癌中高表达,与患者肿瘤分期、分化程度、淋巴结转移、达到满意减瘤术、CA125以及HE4水平有关,与卵巢癌患者的复发与化疗耐药无关,SNCG蛋白的过表达可作为HGSOC患者的独立预后因素,指导临床诊治。

Objective To detect the expression of SNCG in ovarian cancer tissue and its clinical significance, to clarify the relationship between the expression of SNCG in human ovarian cancer and the degree of malignancy, so as to provide theoretical basis for the diagnosis, treatment and prognosis of clinical ovarian cancer. Methods From January 2010 to January 2015 in First Affiliated Hospital of Medical College of Shihezi University,119 patients with ovarian cancer and 50 patients with normal ovarian which had complete clinical data and paraffin section were selected. Immunohistochemical method was used to detect ovarian SNCG expression, and the expression of SNCG relationship with clinical pathological characteristics and prognosis of patients with ovarian cancer was analyzed. Results The expression of SNCG in ovarian cancer tissue was significantly higher than that in normal ovarian tissue (χ²=73.575,P＜0.001). SNCG expression was correlated with tumor stage, degree of differentiation, lymph node metastasis, satisfying tumor reduction, CA125 and HE4 levels in ovarian cancer patients, and the differences were statistically significant (P＜0.05). It was not correlated with the tumor primary site, ascites, recurrence of ovarian tumor and chemotherapy resistance, and the differences were not statistically significant (P＞0.05).The overexpression of SNCG was correlated with PFS and OS in HGSOC patients, and the differences were statistically significant (P＜0.05). Analysis of multivariate Cox proportional risk model showed that SNCG was an independent prognostic factor in patients with HGSOC, related to PFS (HR=2.107,95%CI: 1.014-3.795,P=0.034) and OS (HR=1.238,95%CI: 0.716-1.928,P=0.047). Conclusion The high expression of SNCG in ovarian cancer is related to tumor stage, degree of differentiation, lymph node metastasis, satisfying tumor reduction, CA125 and HE4 expressions, but it is not related to the recurrence of ovarian cancer or chemotherapy resistance. The overexpression of SNCG protein can be used as an independent prognostic factor in patients with HGSOC for clinical diagnosis and treatment guidance.

目的 探讨先天性肥厚性幽门狭窄(congenital hypertrophic pyloric stenosis,IHPS)中神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)的表达情况,以进一步研究IHPS的发病机制。方法 选取4只Beagle孕犬按3:1分为模型组和对照组,自孕期第1天至分娩前1天,模型组孕犬每天腹腔注射一次L-NAME 6 mg/(kg.d),对照组给予同样方式注射等量的生理盐水。所有新生犬均在生后第31天、38天、45天及52天分别称重,生后52天,选取模型组中体质量增长缓慢的新生犬和对照组新生犬各5只作为实验对象,取幽门组织HE染色后显微镜下测量幽门环肌厚度,免疫组化后运用图像分析技术测定nNOS表达量。结果 与对照组新生犬比较,模型组中新生犬体质量增长慢,幽门环肌增厚,nNOS表达量减少,两组间有差异(P<0.01,P<0.01,P<0.05)。结论 幽门区nNOS表达减少与CHPS相关。

Objective To further study the pathogenesis of infantile hypertrophic pyloric stenosis(IHPS) by analyzing the expression of neuronal nitric oxide synthase(nNOS) in pylorus of CHPS. Methods According to 3:1,four Beagle pregnant dogs were selected and divided into model group and control group,from the first day of pregnancy to the first day before delivery, L-NAME was intraperitoneally injected into pregnant dogs in the model group, dose for 6mg/(kg.d),while normal saline was intraperitoneally injected into pregnant dogs in the control group.The two groups newborn dogs were weighed at 31, 38, 45 and 52 days after birth,At 52 days after birth, 5 newborn dogs were selected respectively between model group and control group as experimental subjects, obtained pyloric tissue for HE staining and measured the muscular thickness of pyloric,The expression of nNOS was determined by image analysis after immunohistochemistry. Results Compared with newborn dogs in the control group, newborn dogs in the model group had slow weight growth, increased pyloric annulus muscle thickness, and decreased expression of nNOS(P<0.01,P<0.01,P<0.05). Conclusion The decreased expression of nNOS in the pyloric region associated with IHPS.

目的 研究结肠癌组织中转录因子KLF8的表达及下调KLF8的表达对结肠癌细胞的影响。方法 收集结肠癌组织和癌旁正常组织,检测KLF8的蛋白含量;培养结肠癌Lovo细胞株,转染KLF8 siRNA后检测细胞侵袭、迁移以及上皮-间质转化(EMT)。结果 结肠癌组织中KLF8的蛋白含量高于癌旁正常组织;转染KLF8 siRNA的结肠癌细胞组迁移距离低于阴性对照组,且侵袭至transwell微孔膜外侧面的细胞数少于阴性对照组;转染KLF8 siRNA的结肠癌细胞组内E-cadherin的表达升高,Vimentin、N-cadherin的蛋白含量低于阴性对照组。结论 结肠癌组织中KLF8的表达量升高,下调结肠癌细胞中KLF8的表达可抑制结肠癌细胞侵袭、迁移及上皮-间质转化过程。

Objective To study the expression of transcription factor KLF8 in colorectal cancer tissue and its effect of downregulation KLF8 on colorectal cancer cell. Methods Collecting cancer tissues and adjacent normal color tissue and detecting the protein level of KLF8. Culturing the colorectal cancer Lovo cell lines and detecting cell invasion, cell migration and epithelial-mesenchymal transition after transfecting of KLF8 siRNA. Results KLF8 was highly expressed in colorectal cancer tissues compared with adjacent normal colon tissue. After transfection of KLF8 siRNA, the migration distance of colorectal cancer cell and the cell population transferred to the lateral surface of transwell microporous membrane were lower than those of negative control siRNA. E-cadherin of KLF8 siRNA group were higher than those of negative control siRNA group. Vimentin and N-cadherin were lower than those of negative control siRNA group. Conclusion The expression of KLF8 in colorectal cancer tissue is elevated;downregulation of KLF8 expression in colorectal cancer cell lines may inhibit cell invasion, cell migration and epithelial-mesenchymal transition processes.

目的 研究NR3C1(核受体亚科3,C组,成员1)又称糖皮质激素受体(GR)表达量对前列腺癌恶性程度的影响及其与前列腺癌生化复发的相关性。方法 通过组织芯片免疫组化染色检测的方法检验NR3C1在不同恶性程度前列腺癌组织的表达情况,结合Taylor数据库分析NR3C1表达水平与前列腺癌临床病理特征关系,再采用Kaplan-Meier法分析NR3C1对前列腺癌生化复发生存率的影响,最后用Cox回归分析临床病理特征与生化复发的相关性。结果 组织芯片免疫组化结果显示NR3C1在Gleason评分低的前列腺癌组织中表达高于Gleason评分高的前列腺癌组织(P=0.028)。结合Taylor公用数据库分析,NR3C1在前列腺癌组织中的表达低于癌旁组织(P＜0.001),NR3C1在Gleason评分低的前列腺癌组织中表达高于Gleason评分高的前列腺癌组织(P=0.005),NR3C1低表达与PSA复发(P=0.028)和转移(P=0.003)相关。Kaplan-Meier结果提示:NR3C1高表达组患者术后的生化复发生存率更高(P=0.043),总体生存率没有明显区别(P=0.872)。单因素分析结果显示:NR3C1(P=0.002),病理分期(P＜0.001),Gleason评分(P＜0.001),是否转移(P=0.012)是前列腺癌生化复发的影响因素。多因素分析结果显示:高Gleason 评分(P=0.017)和转移(P<0.001)均为生化复发危险因素。结论 NR3C1影响前列腺癌的发病进程,检验NR3C1的表达情况,能预测前列腺癌患者生化复发的概率,可协助判断前列腺癌预后。

Objective We study the role of NR3C1 (nuclear receptor subfamily 3,group C,member 1) in PCa progression,and the correlation between its expression level and the biochemical recurrence of PCa. Methods Immunohistochemistry was used to detect the expression of NR3C1 in PCa tissues of different degrees of malignancy. The associations of NR3C1 expression and clinical pathological features were analyzed using the Taylor dataset. Kaplan-Meier was used to detect the relationship between NR3C1 expression and biochemical recurrence survival rate in PCa. Cox-regressive analysis was used to detect the relationship between clinical pathological features and biochemical recurrence. Results Immunohistochemistry analysis showed the expression of NR3C1 was higher in which its Gleason Score was lower(P=0.028). Base on the Taylor dataset,the expression of NR3C1 was higher in the adjacent benign tissues than that in PCa(P＜0.001). The expression of NR3C1 was higher in which its Gleason Score was lower(P=0.005). Furthermore,low NR3C1 expression was associated with PSA failure(P=0.028) and Metastasis(P=0.003). Kaplan-Meier showed the biochemical recurrence-free time of PCa patients in low NR3C1 expression groups reduced(P=0.043). The overall survival time of PCa patients was not correlated to NR3C1 expression levels(P=0.872). Single factor analysis showed the biochemical recurrence is associated with NR3C1 expression(P=0.002),pathological stage(P＜0.001),Gleason score(P＜0.001), Metastasis status(P=0.012). Multivariate analysis by Cox regression further identified the high Gleason Score(P=0.017) and Metastasis status (P<0.001)were hazards of the biochemical recurrence. Conclusion Our study showed that the expression of NR3C1 critically connected with the process of PCa,which indicated that we can predict the probability of the biochemical recurrence and determine the prognosis of prostate cancer by detecting the expression of NR3C1 in PCa patients.

目的 分析人脑胶质瘤组织中O6—甲基鸟嘌呤—DNA甲基转移酶(MGMT)、DNA拓扑异构酶Ⅱ(TopoⅡ)基因的表达情况及其对化疗敏感性的影响。方法 收集医院2012年4月—2018年6月期间进行开颅手术切除的新鲜人脑胶质瘤标本80例和同期经颅脑手术治疗的脑外伤或脑出血内减压切除的正常脑组织30例。采用免疫组化法检测两种标本中MGMT和TopoⅡ基因表达情况,并通过脑胶质瘤U251和U87细胞培养、体外药物(替莫唑胺)干预、Transwell体外侵袭实验分析其对化疗敏感性的影响。结果 胶质瘤标本、正常脑组织MGMT和TopoⅡ基因表达程度分布比较差异均有统计学意义(P＜0.05),且二者MGMT基因阳性表达率分别为63.75%、3.33%,TopoⅡ基因阳性表达率分别为55.00%、0.00%,差异有统计学意义(P＜0.05)。MGMT、TopoⅡ基因均在细胞核显示为阳性染色。体外药物干预的实验组、未进行药物干预的阴性对照组干预前U251、U87细胞穿膜细胞计数比较无统计学意义(P＞0.05),但干预后实验组U251、U87细胞穿膜细胞计数高于阴性对照组(P＜0.05),干预后实验组U251、U87细胞有更强的侵袭力。结论 MGMT和TopoⅡ基因在人脑胶质瘤标本中阳性表达率高,且可能参与促进脑胶质瘤细胞侵袭,影响肿瘤化疗敏感性。

Objective To analyze the expression of O6-methylguanine-DNA methyltransferase (MGMT) and DNA topoisomerase II (Topo II) genes in human brain gliomas and its influence on the chemosensitivity. Methods A total of 80 fresh human brain glioma specimens resected by craniotomy and 30 normal brain tissues resected by craniocerebral operation for traumatic brain injury or decompression for cerebral hemorrhage during the period from April 2012 to June 2017 were collected. The expression of MGMT and Topo II genes in the two kinds of specimens was detected by immunohistochemical method, and the influence on chemosensitivity was analyzed through brain glioma U251 and U87 cell culture, in vitro drug (temozolomide) intervention and Transwell invasion in vitro. Results There was a significant difference in the expression of MGMT and Topo II genes in glioma specimens and normal brain tissues (P<0.05). The positive expression rates of MGMT gene in the two kinds of specimens were 63.75% and 3.33% respectively while the positive expression rates of Topo II gene were 55.00% and 0.00%, respectively (P<0.05). Both of MGMT and Topo II genes were displayed in the nucleus as positive staining. There was no significant difference in transmembrane cell count of U251 and U87 cells between the experimental group given drug intervention in vitro and negative control group without drug intervention before the intervention (P>0.05). However, the transmembrane cell count of U251 and U87 cells in the experimental group after intervention was higher than that in the negative control group (P<0.05). After intervetion, U251 and U87 cells were with stronger invasiveness in the experimental group. Conclusion The positive expression rates of MGMT and Topo II genes are high in human brain glioma specimens. They may be involved in promoting glioma cell invasion, affecting tumor chemosensitivity.

目的 探究叶酸对子宫内膜癌作用的靶基因。方法 通过转录组测序筛选叶酸作用下子宫内膜癌细胞中的差异基因,生存分析寻找对子宫内膜癌具有生存意义的差异基因,qPCR及western blot检测其在叶酸作用下的表达。结果 转录组测序发现36个差异基因,生存分析发现FMN1,TRIB3,INHBE及NRBP2的表达对子宫内膜癌具有生存意义,qPCR及western blot验证叶酸作用下NRBP2在子宫内膜癌细胞中的表达下调。结论 叶酸下调子宫内膜癌中NRBP2基因的表达,NRBP2可能是叶酸对子宫内膜癌作用的靶标。

Objective To explore the target genes of folic acid on endometrial carcinoma. Methods The differential genes in endometrial cancer cells treated with folic acid were screened by transcriptome sequencing. Survival analysis was used to find the differential genes with survival significance. QPCR and western blot were used to detect their expression under the action of folic acid. Results 36 differential genes were found by transcriptional sequencing. Survival analysis showed that the expression of FMN1,TRIB3,INHBE and NRBP2 had survival significance in endometrial carcinoma. QPCR and western blot confirmed that the expression of NRBP2 in endometrial cancer cells was down-regulated by folic acid. Conclusion Folic acid down-regulates the expression of NRBP2 gene in endometrial carcinoma, and NRBP2 may be the target of the effect of folic acid on endometrial carcinoma.

目的 应用iTRAQ联合质谱技术筛选COPD大鼠肺组织差异表达蛋白。方法 20只雄性SD大鼠(200~220 g),随机分为对照组和模型组,每组10只,采用熏烟法建立COPD大鼠模型。观察大鼠肺组织病理学改变,测定肺功能,BALF白细胞数,肺组织总蛋白iTRAQ标记后质谱鉴定,用生物信息学方法分析蛋白表达变化。结果 与对照组相比,模型组大鼠支气管黏膜下肌层增厚,肺内可见大量炎性细胞浸润,肺功能降低,BALF白细胞数升高(均P<0.05)。质谱鉴定出4 916种蛋白,筛选出468个差异表达蛋白,其中285个表达上调,183个表达下调。筛选了上皮细胞粘着连接蛋白、fMLP、整合素等与COPD相关蛋白。结论 基于iTRAQ技术的蛋白质组学方法筛选出COPD大鼠差异表达蛋白,为进一步研究COPD的发生机制奠定了基础。

Objective iTRAQ and mass spectrometry were used to screen the differentially expressed proteins in the lung of COPD rats. Methods 20 male SD rats (200-220g)were randomly divided into control group and treatment group, with 10 rats in each group. COPD rat model was established by smoking. The lung function, the number of BALF leukocytes, the total protein iTRAQ in lung tissue were measured and identified by mass spectrometry. The differentially expressed proteins were identified by bioinformatic analysis. Results Compared with the control group, the submucous layer of bronchus in the model group was thickened, a large number of inflammatory cells were seen in the lung, the lung function was reduced, and the number of BALF leukocytes was increased. 4 916 proteins were identified by mass spectrometry, 468 differentially expressed proteins were screened, 285 of which were up-regulated and 183 down regulated. Among them, the important COPD related proteins were epithelial adhesion connexin, fMLP and integrins. Conclusion iTRAQ technology screened out the differentially expressed proteins of COPD rats, which laid the foundation for the further study of COPD mechanism