The regulation of PKCalpha in eosinophil infiltration and proliferation in nasal polyps]

OBJECTIVE: To explore the regulation of protein kinase C (PKC) isoform--PKCalpha in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHOD: With the methods of in situ hybridization staining and immunohistochemistry MGG staining, to check out the relationship between PKC and bcl-2/BaxmRNA and associated protein, especially PKC isoform--PKCalpha, PKCbeta1 , PKCbeta2, and PKCgamma did not express at all. RESULT: There were PKC expression in the eosinophils of 26 cases from nasal polyps, and the expression of PKC and Bcl-2 mRNA/their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01), but in PKC isoforms, PKCalpha expression was the strongest, but PKCbeta1 and PKCbeta2 expressed thinner and PKCgamma did not express at all. CONCLUSION: The reason of eosinophil proliferation and infiltration in nasal polyps is that PKC signal transduction pathway was activated, and leaded to inhibition of eosinophil apoptosis, and eosinophil survival was delayed, and eosinophil proliferated and infiltrated, and in PKC family, PKCalpha is main.     

鼻息肉组织嗜酸粒细胞中水通道蛋白-1和抗凋亡基因Bcl-2 mRNA的表达及意义

OBJECTIVE: To study the significance of aquaporin-1 (AQP-1) expression in the eosinophils of nasal polyps. The expression and location of AQP-1 mRNA and apoptosis associated gene Bcl-2 mRNA in nasal polyps were explored. METHODS: Sixteen nasal polyp samples were collected from 11 women and 5 men aged 20-65 years during routine endonasal surgery. Nasal mucosa specimens from the inferior turbinates of 10 patients with allergic rhinitis (7 women and 3 men, aged 16-58 years), collected during septoplasty, were used as controls. The expression of AQP-1 mRNA and Bcl-2 mRNA was detected in serial adjacent sections by in situ hybridization and eosinophils were examined by stain MGG. RESULTS: AQP-1 mRNA expression was found in all 16 nasal polyps and in 4 of 10 inferior turbinate tissues, the mean expression rates were (93.16 +/- 13.25)% and (19.54 +/- 4.98)%, respectively. All 16 nasal polyps and 10 control nasal tissues expressed Bcl-2 mRNA, by the average rates of (84.74 +/- 12.10)% and (16.45 +/- 3.12)%, respectively. The expression of AQP-1 mRNA was positively correlated with Bcl-2 mRNA expression in nasal polyps (r = 0.875, P < 0.01). CONCLUSIONS: AQP-1 contributes to the survival of eosinophils in nasal polyps by keeping the permeation balance of eosinophils.     

Effect of erythromycin on the expression of apoptotic gene on eosinophils in nasal polyps in vitro]

OBJECTIVE: To study the expression of apoptotic gene in nasal polyps and the possible effect of erythromycin on it. METHODS: Total 40 nasal polyps biopsies were detected the expression of Bcl-2, Bax, Bcl-xL as well as Fas protein on eosinophils by immunohistochemistry. The expression of Bax, Bcl-xL, Fas on eosinophils in 15 nasal polyps explants which cultured with or without erythromycin were detected also. RESULTS: In nasal polyps, the expression of Bcl-2, Bax, Bcl-xL and Fas protein on eosinophils are 5% (2/10), 65% (26/40), 40% (16/40) and 90% (36/40) respectively. As compared with inferior turbinate, there were significant difference in expression of Bax and Fas protein between two tissues (P < 0.05). After cultured with erythromycin, the expression of Bax protein increased (P < 0.05), but the expression of Bcl-xL and Fas protein had not changed. CONCLUSION: Bax may be the most significant gene which control the apoptosis of eosinophils in nasal polyps. And a possible mechaism which erythromycin promote the apoptosis of eosinophils is increase the expression of Bax.     

Expression of cell proliferation and apoptosis gene associated protein on nasal polyps and its significance]

OBJECTIVE: To further understand the pathogenic mechanism of nasal polyps, namely the cells proliferation and apoptosis in nasal polyps tissue. METHODS: The proliferation cell nuclear antigen (PCNA), apoptosis associated gene protein (Bcl-2, Bax) were determined in 26 tissue samples of nasal polyps and 14 controls from normal inferior turbinates respectively. RESULTS: (1) The positive expression e of PCNA, Bcl-2 and Bax were significantly higher in epithelium of nasal polyps than the controls, and the expression rate of Bcl-2/Bax, however, tended equipoise. (2) The expression of Bcl-2 were significantly stronger than Bax in glands, eosinophils of nasal polyps tissue, and there were not significant differences in inferior turbinates. CONCLUSION: There is strongly proliferation activity in the epithelium of nasal polyps, and expression imbalance of Bcl-2/Bax may be one of the important factors of eosinophilia in nasal polyps tissue.     

鼻息肉中IL-15的含量及其与Bcl-2、Bcl-XL表达的关系

目的：观察IL-15在鼻息肉与下鼻甲中表达的差异及其与Bcl-2、Bcl-XL表达的关系,探讨IL-15在鼻息肉发病机制中的作用。方法用ELISA的方法测定24例鼻息肉与8例下鼻甲的IL-15含量,同时用免疫组化法检测其组织中Bcl-2及Bcl-XL的表达情况。结果鼻息肉组中IL-15表达高于下鼻甲组,差别有统计学意义（P＜0.05）。鼻息肉组中Bcl-XL表达高于下鼻甲组,差别有统计学意义（P＜0.01）;Bcl-2的表达两组间差别无显著性;IL-15的表达与Bcl-2和Bcl-XL的表达之间没有相关性。结论在鼻息肉发病过程中,IL-15在嗜酸性粒细胞等炎症细胞的凋亡抑制过程中可能扮演着重要的角色。     

红霉素干预对人鼻息肉上皮细胞凋亡的影响

目的观察红霉素对体外培养的鼻息肉上皮细胞B细胞淋巴瘤-2基因(B cell lymphomas-2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)表达的影响,探讨上皮细胞凋亡在鼻息肉组织形成中的意义。方法将30例鼻息肉及15例下鼻甲组织均分成两组,分别对其上皮细胞进行原代培养,其中一组加入红霉素。培养第1、3、5天应用末端脱氧核苷酰转移酶介导的脱氧尿苷三磷酸原位缺口末端标记法(the TdT-mediated dUTP nick end labeling,TUNEL)检测细胞凋亡情况,并用免疫组化法检测细胞凋亡基因Bax、抑凋亡基因Bcl-2的表达。结果①鼻息肉红霉素组培养第1、3、5天的上皮细胞凋亡指数(用x±s表示,下同)分别为(33．23±6．50)％、(36．14±3．21)％和(52．63±7．86)％,对照组(不加红霉素组)为(31．02±5．60)％、(33．88±4．02)％和(39．64±7．48)％,培养第5天的上皮细胞凋亡指数红霉素组与对照组差异有统计学意义(t=4．480, P<0．05)。②鼻息肉上皮细胞Bcl-2和Bax表达均显著强于下鼻甲组织(P<0．01);培养第5天鼻息肉上皮细胞红霉素组Bax表达显著强于对照组(t=8．734,P<0．05),Bcl-2表达虽强于对照组,但差异无统计学意义;下鼻甲上皮细胞红霉素组与对照组的Bcl-2与Bax阳性表达差异均无统计学意义(P均>0．05);红霉素干预第5天鼻息肉上皮细胞显示明显的凋亡趋势(P<0．05)。结论红霉素能显著提高Bax蛋白的表达,有显著的促鼻息肉上皮细胞凋亡的作用。     

鼻息肉中诱导型一氧化氮合酶、Bax及Bcl-2的表达及相关性研究

目的:检测鼻息肉组织中诱导型一氧化氮合酶(iNOS)和凋亡相关基因Bax、Bcl-2的表达及相关性。方法:30例鼻息肉组织,应用TUNEL方法检测其凋亡情况;免疫组织化学SABC法检测iNOS及Bax、Bcl-2的表达;western blot方法检测iNOS、Bax、Bcl-2的表达。结果:①TUNEL方法检测凋亡细胞位于鼻息肉表层上皮细胞及腺体上皮细胞,均为弱阳性。②免疫组织化学法检测iNOS、Bax、Bcl-2均表达于鼻息肉组织上皮细胞、腺上皮细胞、血管内皮细胞和浸润的炎性细胞,阳性表达定位于细胞质,其中Bax表达为弱阳性,iNOS、Bcl-2表达为强阳性。③Western blot检测3种蛋白均有表达条带,iNOS、Bal-2条带清晰,Bax条带软弱,差异有统计学意义(P<0.01);iNOS与Bcl-2的灰度值比值呈正相关(r=0.851,P<0.01);iNOS与Bax的灰度值比值呈负相关(r=-0.714,P<0.01)。结论:鼻息肉组织中存在抗凋亡和促凋亡2种蛋白,iNOS抑制细胞凋亡,可能对鼻息肉的发生、发展起重要的促进作用。     

鼻息肉中单核细胞趋化蛋白1和血管内皮生长因子的表达

目的明确单核细胞趋化蛋白1（monocyte chemotactic protein 1，MCP-1）和血管内皮生长因子（vascular endothelial growth factor，VEGF）在鼻息肉组织中的表达及其相关性，初步探讨MCP-1与鼻息肉发生的关系。方法取40例鼻息肉组织和25例下鼻甲组织，应用原位杂交和免疫组织化学等方法检测MCP-1和VEGF mRNA及蛋白质的表达。结果鼻息肉组织中MCP-1和VEGF mRNA及蛋白质的表达均高于对照组下鼻甲组织（P值均〈0．01）；鼻息肉组织中MCP-1和VEGF蛋白质的表达呈正相关（r=0．871，P〈0．05）。结论鼻息肉组织中MCP-1和VEGF表达增加，二者协同作用可能是鼻息肉形成的原因之一。     

黏附分子及血管内皮生长因子在鼻息肉中的表达

目的 探讨细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）以及血管内皮生长因子（VEGF）在鼻息肉中的表达与意义。方法25例鼻息肉标本和9例下鼻甲黏膜标本，分别行ICAM-1、VCAM-1、VEGF免疫组化染色和HE染色，光镜下观察比较各种分子表达水平。结果 鼻息肉中可见大量嗜酸性粒细胞（EOS）浸润。ICAM-1、VCAM-1和VEGF均可表达于鼻息肉血管内皮、间质及炎症细胞，且在鼻息肉血管内皮、间质及浸润炎症细胞中的表达趋势呈现正相关关系。结论 ICAM-1、VCAM-1可能与EOS等炎症细胞附壁浸润活化过程关系密切，VEGF可能启动并加强这一病理变化。它们的协同作用可能参与了鼻息肉的病理过程。     

蛋白激酶C抑制剂对离体鼻息肉组织中嗜酸粒细胞凋亡的影响

OBJECTIVE: To study the influence of the inhibitor of protein kinase C (PKC), Iso-H7 [1-(5-lsoquinolinyl-sulfony)-3-methylpiperazine dihydrochloride] on eosinophil apoptosis in nasal polyp tissues. METHODS: Nasal polyp tissues were cultured and divided into two groups, test group (with Iso-H7 in different concentration) and control group (without the inhibitor of PKC), to check out the influence of Iso-H7 on eosinophil apoptosis in nasal polyps. The nasal polyp tissues were cultured in Iso-H7 in concentration of 10(-6), 10(-4), 10(-2) mol/L for 1 d, 3 d, and 5 d, respectively. Eosinophil apoptosis index was measured with TUNEL (terminal-deoxynucleotidyl transferase d-UTP nick end labeling), in situ hybridization and MGG staining to identify eosinophil apoptosis. RESULTS: Occasionally, there was little eosinophil apoptosis in two groups in 1 d, but in 3 d, There were a great deal of eosinophil apoptosis in test group. The eosinophil apoptosis indexs of the two groups were significantly different (P < 0.01). The indexs of eosinophil apoptosis was not significantly higher in 3 d than that in 5 d in test groups (P > 0. 05), but significantly higher than that in 1d (P < 0. 01). CONCLUSIONS: Iso-H7 takes a important role in accelerating eosinophil apoptosis. The inhibitor of PKC could significantly promote eosinophil apoptosis in nasal polyps. There is no dosage and time dependence of PKC inhibitor in regulating eosinophil apoptosis of nasal polyps.     

The concentration and expression of IL-5 in human nasal polyp tissue]

OBJECTIVE: To study the concentration and expression of IL-5 in nasal polyp tissues and explore its significance in the micro-environment differentiation of eosinophils accumulation and clarify the conception of nasal polyposis. METHODS: The concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry, and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers was used as control. RESULTS: 1. IL-5 concentration in the polyp tissues was significantly higher than that in inferion turbinate mucosa(P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentration in polyp tissues was markedly higher in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). IL-5 concentration had no correlation with age and sex (P > 0.05). 2. 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of the lymphocytes and neutrophils were IL-5 positive, and IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissues was significantly stronger in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and inferior turbinate nasal mucosa (both P > 0.05). CONCLUSION: IL-5 is a key protein in eosinophilic pathologic mechanisms in nasal polyp tissues.     

胰岛素样生长因子1在鼻息肉组织中的表达与IgE的相关性研究

目的探讨胰岛素样生长因子1(insulin-likegrowth factor 1,IGF-1)在鼻息肉组织中的表达及其与IgE的相关性。方法使用荧光定量PCR技术和免疫组织化学技术分别检测40例伴鼻息肉的慢性鼻及鼻窦炎(chronic rhinosinusitis with nasal polyps,CRSwNP)患者的鼻息肉组织(CRSwNP组)和20例鼻中隔偏曲患者的中鼻甲组织(对照组)中IGF-1 mRNA及其蛋白的表达、免疫组织化学技术检测所有标本的总IgE表达并进行统计学分析。结果 IGF-1蛋白在CRSwNP组及对照组的阳性率分别为75.3%和15.4%,IGF-1 mRNA在两组的表达分别为0.825±0.315和0.358±0.387,CRSwNP组IGF-1蛋白和IGF-1 mRNA的表达显著高于对照组(t=6.318,P<0.01;t=-5.014,P<0.05);CRSwNP组中合并变应性因素与无变应性因素亚组之间的IGF-1 mRNA表达无差异(t=0.856,P>0.05)。总IgE在两组阳性细胞数分别为14.65±3.34和3.40±1.27,CRSwNP组总IgE阳性细胞数显著高于对照组(t=-5.366,P<0.01)。IGF-1 mRNA与总IgE阳性细胞数呈正相关(r=0.319,P<0.05)。结论IGF-1参与CRSwNP的发病机制,局部变应性因素而非系统性变应性因素影响其表达,IGF-1可能通过促进细胞生长、抑制细胞凋亡以及促进IgE的生成参与局部免疫调控等途径发挥作用。     

NLRP3炎性小体在嗜酸粒细胞性鼻息肉发病及复发中的作用

目的 探讨NLRP3炎性小体在嗜酸粒细胞性鼻息肉(ENP)发病及复发中的可能作用及机制。 方法 根据HE染色,选取80例ENP(ENP组)、24例嗜酸粒细胞性复发鼻息肉(RENP)患者(RENP组)的鼻息肉组织及同期30例鼻中隔偏曲患者(对照组)的中鼻甲黏膜。所有患者嗜酸粒细胞(EOS)浸润数通过HE染色观察并计算,采用免疫组化法检测3组NLR含热蛋白结构域3(NLRP3)炎性小体、白细胞介素-1beta(IL-1β)、IL-18的表达情况,并分析ENP、RENP组中NLRP3炎性小体与EOS浸润数的相关性。 结果 IL-1β和IL-18在ENP及RENP组中的表达均高于对照组(P<0.05),但在ENP与RENP组中的表达无显著差异(P>0.05)。NLRP3炎性小体在RENP组中的表达明显高于ENP组和对照组(P<0.01),且EOS浸润数明显多于ENP组(P<0.05)和对照组(P<0.01)。NLRP3炎性小体在ENP及RENP组中的表达均与EOS浸润数呈正相关。 结论 NLRP3炎性小体、IL-1β和IL-18均与ENP发病密切相关,NLRP3炎性小体可能参与ENP的复发。     

The protein expression of Bcl-x1 and Bcl-2 in eosinophils in nasal polyps and the influence of beclomethasone dipropionate on the expression]

OBJECTIVE: To investigate the protein expression of B-cell lymphoma/leukemia-2(Bcl-2) and B-cell lymphoma/leukemia-x long(Bcl-x1) in eosinophils in nasal polyps and the influence of beclomethasone dipropionate on the expression. METHODS: Using May-Grünwald-Giemsa (MGG) method and immunohistochemical method, the protein expression of Bcl-x1 and Bcl-2 in eosinophils in nasal polyps from patients treated with beclomethasone dipropionate treatment and patients without any treatment was compared. RESULTS: (1) Nasal polyp tissues from patients without treatment had more eosinophils than those from patients with treatment(P < 0.01). (2) No protein expression of Bcl-2 was observed in all 52 patients. (3) 20.0% patients with treatment had the expression of Bcl-x1, whereas 48.1% patients without treatment had the expression. The difference between these two groups was significant(P < 0.05). CONCLUSION: These data suggest that Bcl-x1 may act as an anti-apoptotic molecule in eosinophils and corticosteroids induce eosinophil apoptosis through suppressing the expression of Bcl-x1.     

鼻息肉中总IgE及IL-5与嗜酸性粒细胞浸润的相关性研究

目的探讨鼻息肉中总IgE、IL-5的表达、嗜酸性粒细胞(eosinophils,EOS)浸润情况及其相关关系,以分析其在鼻息肉发病中的作用.方法采用免疫组化SP法检测38例鼻息肉,15例正常鼻腔黏膜组织中总IgE阳性细胞分布情况;酶联免疫吸附试验(ELISA)检测鼻息肉中IL-5含量;Chromotrope 2R特染法检测鼻息肉中EOS浸润情况.结果①鼻息肉组织中总IgE、IL-5及EOS水平均明显高于正常鼻腔黏膜组织,P均＜0.01;②鼻息肉组织中总IgE与EOS、IL-5与EOS以及IL-5与总IgE之间均呈明显正相关关系,相关系数r分别为0.843,0.642,0.667,P均＜0.01.结论EOS浸润在鼻息肉的形成和发展中起重要作用,而IgE及IL-5则对鼻息肉中EOS浸润发挥重要影响.     

天然免疫蛋白PLUNC在鼻息肉中的表达及与鼻息肉临床特征的关联性

目的：探讨天然免疫蛋白腭、肺、鼻上皮克隆（PLUNC）在鼻息肉组织中的表达情况,分析PLUNC蛋白浓度与鼻息肉大小（鼻内镜记分）和手术后复发的关联性。方法：采用免疫组织化学和实时定量PCR检测鼻息肉组（28例鼻息肉患者,其中13例为术后复发患者）和对照组（16例钩突黏膜对照）PLUNC的组织染色定位和mRNA的表达情况,同时采用ELISA检测初发和复发鼻息肉组织中PLUNC的蛋白浓度差异,评估PLUNC蛋白浓度与息肉大小、鼻塞、流涕症状记分的相关性。结果：鼻息肉组织中PLUNC主要定位在黏膜上皮和腺体,染色强度记分显著低于对照的钩突组织（P〈0．01）;PLUNCmRNA表达水平也显著低于对照的钩突组织（P〈0．01）。初发和复发鼻息肉组织中PLUNC蛋白OD值分别为0．33±0．11和0．15±0．05,差异有统计学意义（P〈0．01）。小型和大型鼻息肉PLUNC蛋白OD值分别为0．32±0．14和0．19±0．07,差异有统计学意义（P〈0．05）。鼻息肉组织中PLUNC蛋白浓度与鼻塞和流涕记分存在显著负相关（r=-0．51和r=-0．57,P〈0．01）。结论：PLUNC的表达降低提示鼻息肉的发生可能与天然免疫反应的减弱有关,因此,上调天然免疫分子如PLUNC等的表达可作为干预鼻息肉发病的一个新策略。     

糖皮质激素受体 mRNA在慢性鼻窦炎鼻息肉组织中的表达

目的通过检测糖皮质激素受体(glucocorticoid receptor, GR)-α及β mRNA在接受鼻腔糖皮质激素治疗后不同疗效的两组患者鼻息肉黏膜的定量表达,探讨在不同组慢性鼻窦炎鼻息肉组织中GR-α和GR-β的表达意义.方法采用荧光定量反转录聚合酶链反应(FQ-RT-PCR)方法检测40例患者鼻息肉组织中GR-α mRNA和GR-β mRNA的定量表达.结果通过随访80例门诊患者后筛查出40例患者,其中激素敏感组26例,激素不敏感组14例.在激素不敏感鼻息肉组织中GR-β mRNA 表达[(5.72±0.58)×102 拷贝/μg]均高于激素敏感组[(4.82±0.28)×102拷贝/μg,t=-6.65,P<0.01]和正常鼻黏膜组[(4.44±0.35)×102拷贝/μg,t=-9.19, P<0.01],并在该两组之间差异均有统计学意义.GR-α mRNA与GR-β mRNA的比值在激素敏感组(829.42±67.36)与激素不敏感组(535.70±89.00)间差异有统计学意义(t=11.74, P<0.01).结论慢性鼻-鼻窦炎患者中的GR-β mRNA高表达,GR-α mRNA表达下调提示激素不敏感的存在,GR-β在评价鼻息肉患者对激素治疗疗效上有一定作用.     

两种细胞外基质糖蛋白在鼻息肉中的表达

OBJECTIVE: To explore the role of tenascin (TN) and fibronectin (FN) in the pathophysiology of nasal polyp. METHODS: The expression of TN and FN in nasal polyps from 34 patients and in inferior turbinates from 20 patients with deviation of nasal septum was studied with immunohistochemical method. In patients with nasal polyps, the relations between expression and histopathologic characteristics, eosinophilias (EOS) infiltration, clinical staging and the size of nasal polyps were analyzed. RESULTS: (1) The gray score of TN and FN expression was 163.10 +/- 10.54 and 163.24 +/- 11.52 in nasal polyps respectively, whereas it was 175.49 +/- 9.29 and 173.93 +/- 7.92 in inferior turbinates respectively. The difference between two groups was significant(P < 0.01); (2) The expression of TN and FN in edematous type was significantly stronger than that in cystic fibrous and glandular type (P < 0.05); (3) The association between FN expression and EOS infiltration was significant(r = -0.60, P < 0.01); (4) The expression of TN and FN did not correlate with clinical staging and size(P > 0.05). CONCLUSION: Abnormal ECM may contribute to proliferation of epithelium, accumulation of EOS and edema formation, as a result, to enhance the development of nasal polyps.