中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2014年
7期
568-573
,共6页
陈曦%郭洁波%张涵%左可军%韦轶%史剑波%李华斌%许庚
陳晞%郭潔波%張涵%左可軍%韋軼%史劍波%李華斌%許庚
진희%곽길파%장함%좌가군%위질%사검파%리화빈%허경
鼻息肉%连接蛋白类%闭锁蛋白%细胞因子类%实时聚合酶链反应
鼻息肉%連接蛋白類%閉鎖蛋白%細胞因子類%實時聚閤酶鏈反應
비식육%련접단백류%폐쇄단백%세포인자류%실시취합매련반응
Nasal polyps%Connexins%Occudin%Cytokines%Real-time polymerase chain reaction
目的 探讨紧密连接蛋白Occludin在鼻息肉发病中的作用.方法 采用免疫组化和实时荧光定量聚合酶链反应检测鼻息肉组织(n=20)和健康对照组钩突组织(n=15)中紧密连接蛋白Claudin-1、Occludin及ZO-1蛋白及其mRNA的表达.并通过离体细胞培养模型检测白细胞介素(interleukin,IL)6、IL-13、IL-17、γ干扰素(interferon-y,IFN-γ)、转化生长因子β(transforming growth factor-β、TGF-β)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)刺激对鼻黏膜上皮细胞表达Occludin mRNA和蛋白的调节作用.以SPSS 16.0软件进行统计学分析.结果 紧密连接蛋白Claudin-1、Occludin和ZO-1在鼻息肉与对照组钩突组织中均有表达,主要表达部位在鼻黏膜上皮细胞的细胞膜与细胞质.鼻息肉组Claudin-1、Occludin及ZO-1的平均吸光度值分别为0.187±0.076、0.172 ±0.109、0.098±0.035,均较对照组的0.312 ±0.101、0.220±0.069、0.233±0.093明显减少,差异有统计学意义(t值分别为9.345、3.301、13.323,P值均＜0.01).鼻息肉组织中Occludin mRNA相对表达量为0.000 117 ±0.000 035,低于对照组的0.000464±0.000 134,差异具有统计学意义(Z=-5.0,P＜0.05),而Claudin-1和ZO-1 mRNA在两组之间表达差异无统计学意义(P值均＞0.05).IL-13、IL-17和IFN-γ等促炎因子刺激鼻黏膜上皮细胞后,Occludin mRNA相对表达量分别为0.631±0.039、0.581±0.029、0.648±0.040,与未刺激对照组相比,差异有统计学意义(f值分别为16.299、24.669、14.995,P值均＜0.05).蛋白免疫印迹结果显示上述因子刺激组与未刺激对照组相比Occludin蛋白表达升高,差异有统计学意义(t值分别为9.880、8.442、7.310,P值均＜0.05).结论 紧密连接蛋白Occludin表达降低可能是鼻息肉发生的原因之一.
目的 探討緊密連接蛋白Occludin在鼻息肉髮病中的作用.方法 採用免疫組化和實時熒光定量聚閤酶鏈反應檢測鼻息肉組織(n=20)和健康對照組鉤突組織(n=15)中緊密連接蛋白Claudin-1、Occludin及ZO-1蛋白及其mRNA的錶達.併通過離體細胞培養模型檢測白細胞介素(interleukin,IL)6、IL-13、IL-17、γ榦擾素(interferon-y,IFN-γ)、轉化生長因子β(transforming growth factor-β、TGF-β)、腫瘤壞死因子α(tumor necrosis factor-α,TNF-α)刺激對鼻黏膜上皮細胞錶達Occludin mRNA和蛋白的調節作用.以SPSS 16.0軟件進行統計學分析.結果 緊密連接蛋白Claudin-1、Occludin和ZO-1在鼻息肉與對照組鉤突組織中均有錶達,主要錶達部位在鼻黏膜上皮細胞的細胞膜與細胞質.鼻息肉組Claudin-1、Occludin及ZO-1的平均吸光度值分彆為0.187±0.076、0.172 ±0.109、0.098±0.035,均較對照組的0.312 ±0.101、0.220±0.069、0.233±0.093明顯減少,差異有統計學意義(t值分彆為9.345、3.301、13.323,P值均＜0.01).鼻息肉組織中Occludin mRNA相對錶達量為0.000 117 ±0.000 035,低于對照組的0.000464±0.000 134,差異具有統計學意義(Z=-5.0,P＜0.05),而Claudin-1和ZO-1 mRNA在兩組之間錶達差異無統計學意義(P值均＞0.05).IL-13、IL-17和IFN-γ等促炎因子刺激鼻黏膜上皮細胞後,Occludin mRNA相對錶達量分彆為0.631±0.039、0.581±0.029、0.648±0.040,與未刺激對照組相比,差異有統計學意義(f值分彆為16.299、24.669、14.995,P值均＜0.05).蛋白免疫印跡結果顯示上述因子刺激組與未刺激對照組相比Occludin蛋白錶達升高,差異有統計學意義(t值分彆為9.880、8.442、7.310,P值均＜0.05).結論 緊密連接蛋白Occludin錶達降低可能是鼻息肉髮生的原因之一.
목적 탐토긴밀련접단백Occludin재비식육발병중적작용.방법 채용면역조화화실시형광정량취합매련반응검측비식육조직(n=20)화건강대조조구돌조직(n=15)중긴밀련접단백Claudin-1、Occludin급ZO-1단백급기mRNA적표체.병통과리체세포배양모형검측백세포개소(interleukin,IL)6、IL-13、IL-17、γ간우소(interferon-y,IFN-γ)、전화생장인자β(transforming growth factor-β、TGF-β)、종류배사인자α(tumor necrosis factor-α,TNF-α)자격대비점막상피세포표체Occludin mRNA화단백적조절작용.이SPSS 16.0연건진행통계학분석.결과 긴밀련접단백Claudin-1、Occludin화ZO-1재비식육여대조조구돌조직중균유표체,주요표체부위재비점막상피세포적세포막여세포질.비식육조Claudin-1、Occludin급ZO-1적평균흡광도치분별위0.187±0.076、0.172 ±0.109、0.098±0.035,균교대조조적0.312 ±0.101、0.220±0.069、0.233±0.093명현감소,차이유통계학의의(t치분별위9.345、3.301、13.323,P치균＜0.01).비식육조직중Occludin mRNA상대표체량위0.000 117 ±0.000 035,저우대조조적0.000464±0.000 134,차이구유통계학의의(Z=-5.0,P＜0.05),이Claudin-1화ZO-1 mRNA재량조지간표체차이무통계학의의(P치균＞0.05).IL-13、IL-17화IFN-γ등촉염인자자격비점막상피세포후,Occludin mRNA상대표체량분별위0.631±0.039、0.581±0.029、0.648±0.040,여미자격대조조상비,차이유통계학의의(f치분별위16.299、24.669、14.995,P치균＜0.05).단백면역인적결과현시상술인자자격조여미자격대조조상비Occludin단백표체승고,차이유통계학의의(t치분별위9.880、8.442、7.310,P치균＜0.05).결론 긴밀련접단백Occludin표체강저가능시비식육발생적원인지일.
Objective To evaluate the possible role of tight junction protein Occludin in nasal polyps.Methods The expression of Claudin-1,Occludin and ZO-1 in nasal polyps (n =20) and healthy uncinate mucosa (n =15) were examined using immunohistochemical staining,real-time quantitative polymerase chain reaction (PCR) and Western blot analysis.The regulatory effects of proinflammatory cytokines (IFN-γ,IL-13,IL-17,TGF-β,TGF-α) on the expression of Occludin in cultured human nasal epithelial cells were investigated.Results The immunohistochemical results showed that Claudin-1,Occludin and ZO-1 were detected both in the nasal polyp group and the control group.The expression sites were the cell membrane and cytoplasm of nasal mucosa epithelial cells.The mean optical density of Claudin1,Occludin and ZO-1 were 0.187 ± 0.076,0.172 ± 0.109 and 0.098 ± 0.035 respectively in the nasal polyp group and were significantly lower than those in the control group (0.312 ± 0.101,0.220 ± 0.069 and 0.233 ± 0.093 respectively),the differences were significant (t =9.345,t =3.301,t =13.323,all P ＜ 0.01).RT-PCR results showed that the relative expression of Occludin mRNA was 0.000 117 ± 0.000 035 in the nasal polyp group and was significantly lower than that in the control group(0.000 464 ±0.000 134),and the difference was significant (Z =-5.0,P ＜ 0.01).There was no statistically significant difference in the relative expression of Claudin-1 and ZO-1 mRNA between the nasal polyp group and the control group(P ＞ 0.05).After the cultured human nasal epithelial cells were stimulated by IL-13,IL-17,IFN-γ and other proinflammatory cytokines,the relative expression of Occludin mRNA was 0.631 ± 0.039,0.581 ± 0.029 and 0.648 ± 0.040,respectively.Compared with the unstimulated control group,the differences were statistically significant (t =16.299,24.669 and 14.995 respectively,all P ＜ 0.05).Western blot analyse showed that the relative grayscale in the above proinflammatory cytokines stimulation groups was 0.650 ± 0.061,0.482 ± 0.106 and 0.536 ± 0.109,respectively.Compared with the unstimulated control group,the differences were statistically significant (t =9.880,8.442 and 7.310 respectively,all P＜0.05).Conclusions The reduced expression of Occludin might be involved in the pathogenesis of nasal polyps.
