TNF-α和IL-1α在中耳胆脂瘤和肉芽组织中的表达及意义
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摘要
目的：通过研究肿瘤坏死因子-α（TNF-α）和白细胞介素-1α（IL-1α）在中耳胆脂瘤和肉芽组织中的表达情况，探讨其在慢性化脓性中耳炎发病机制中的作用。
方法：选取山西医科大学第一临床医学院耳鼻咽喉-头颈外科行慢性化脓性中耳炎乳突根治手术的81例患者，其中胆脂瘤标本35例，肉芽组织标本20例，包含胆脂瘤和肉芽组织的标本26例，上述病例均为单侧耳发病，详细记录患者的年龄、性别、发病时间、听力损失程度及术中所见骨质破坏程度，选取乳突根治术患者正常外耳道上皮组织10例作为对照，所有标本均经10%中性甲醛固定，常规石蜡包埋，采用免疫组化二步法和计算机图像定量分析系统测定TNF-α和IL-1α分别在上述三型标本中的表达情况。
结果：
1. TNF-α和IL-1α在中耳胆脂瘤中的表达阳性率分别为89%和86%，经χ~2连续性校正检验，与正常外耳道上皮对比差异有统计学意义（P<0.05）。TNF-α在正常外耳道上皮的阳性表达表现为镜下胞浆可见淡棕色颗粒，密度低，位于上皮表层；胆脂瘤中TNF-α阳性表达位于上皮全层，以基底层密度最高，镜下胞浆呈现密集棕黄色颗粒，IL-1α在外耳道上皮中呈弱阳性表达，仅在基底层可见散在的阳性细胞；在胆脂瘤中的阳性表达位于上皮全层，以基底层和棘层密度最高，镜下胞浆呈现高密度的棕黄色颗粒。
2. TNF-α和IL-1α在肉芽组织中的表达阳性率分别为85%和80%，经Fisher确切概率法检验，与正常外耳道上皮对比差异有统计学意义（P<0.05），两者在肉芽组织中的表达大多位于中性粒细胞和巨噬细胞中，且血管内皮细胞显示中等阳性，镜下胞浆可见高密度棕黄色颗粒。
3. TNF-α和IL-1α在包含有胆脂瘤和肉芽组织的标本中阳性表达率分别为96%和88%，镜下可见阳性细胞表达从基底层向角质层逐渐增强，肉芽组织周围和棘细胞层染色较深。
4.经单因素方差分析可得出：包含有胆脂瘤和肉芽的组织TNF-α的表达程度高于仅有肉芽的组织和仅有胆脂瘤的组织（P<0.01），仅有胆脂瘤的组织表达高于仅有肉芽的组织（P<0.01），IL-1α在包含有胆脂瘤和肉芽的组织中表达程度高于仅有肉芽的组织和仅有胆脂瘤组织（P<0.01），在仅有胆脂瘤的组织中的表达高于仅有肉芽的组织（P<0.01）。
5.含有胆脂瘤的组织中TNF-α和IL-1α两者的表达呈正相关(r=0.972，P<0.01)，在肉芽组织中TNF-α和IL-1α的表达亦呈正相关（r=0.974，P<0.01)；胆脂瘤和肉芽组织骨质破坏力与TNF-α和IL-1α的表达有显著相关性(均为P<0.01)。TNF-α和IL-1α表达灰度值越低,表达密度越高,胆脂瘤和肉芽组织的骨质破坏力越强。
结论：
1. TNF-α、IL-1α在成人中耳胆脂瘤和肉芽组织中呈高表达，说明两者在胆脂瘤和肉芽组织形成中起着重要作用，TNF-α和IL-1α的表达有相关性，提示二者的形成相互影响。
2. TNF-α和IL-1α在包含有胆脂瘤和肉芽的组织中表达程度高于仅有肉芽和仅有胆脂瘤的组织，说明胆脂瘤和肉芽组织形成过程中局部始终处于炎症状态。
3.中耳骨质破坏程度与TNF-α和IL-1α的表达有显著相关性，提示TNF-α和IL-1α可能与胆脂瘤和肉芽组织的骨质破坏机制密切相关。
4. TNF-α和IL-1α在胆脂瘤和肉芽组织的高表达与胆脂瘤的骨质破坏及增殖能力具有非常重要的作用，为进一步揭示胆脂瘤的病理分子机制提供了实验基础。
5. TNF-α和IL-1α在胆脂瘤和肉芽组织的异常表达状态，对全面了解胆脂瘤和肉芽组织的生物学行为奠定理论基础。
Object To study the expression of tumor necrosis factor-alpha (TNF-α) andinterleukin -1-alpha(IL-1α) in human middle ear cholesteatoma and granulation tissuefor discussing their roles in the otitis media development.
Methods The specimen are selected from the First Clinical Medical College ofShanxi Medical University, department of Otolaryngology-Head and Neck , including35 cases from cholesteatoma specimens, 20 cases from granulation tissue specimensand 26 cases from both including cholesteatoma and granulation tissuespecimens.Write down their age, gender, illness duration, degree of hearing loss,andobserve their bone damage by surgery meanwhile.In addition, patients with normalexternal ear canal epithelium in 10 cases as controls. Finally,all specimens were fixedin 10% neutral formalin and paraffin-embedded.To measure the expression of TNF-αand IL-1αin the middle ear cholesteatoma and granulation tissue byimmunohistochemical technology and computer image analysis system.
Results
1. The percent of positive expression of TNF-αand IL-1αin middle earcholesteatoma were 89% and 86%, as compared with the normal ear canal skin,there were significant differences in expression (P<0.05.). The positive expressionof TNF-αin normal external ear canal epithelium cytoplasm showed a light browngranular and lowly density,which have a higher dendity expression in allcholesteatoma epithelial layers, the highest density in the basal layer, subepithelialconnective tissue is also have a positive expression.There is a weakly expressionof IL-1αin the external auditory canal epithelium and the positive cells onlyscattered in the basal layer,which have a higher expression in the wholecholesteatoma layer, with the highest density in basal and spinous layer.
2. The percent of positive expression of TNF-α, and IL-1αin granulation tissue were85% and 80%, as compared with the normal ear canal skin, there were significantdifferences in expression (P<0.05.).Both have a higher expression in neutrophilegranulocyte and macrophages,also in the vascular endothelial cells.
3. The percent of positive expression of TNF-αand IL-1αin the specimen includingcholesteatoma and granulation tissue were 96% and 88%,by the microscope we could observed that positive cells gradually increased from the basal layer to thestratum corneum,particularly have a higher expression around the granulationtissue and stratum spinosum epidermidis.
4. By the Oneway ANOVA analysis can observed :The expression of TNF-αin thespecimen including cholesteatoma and granulation tissue have a higher level thanthe granulation tissue and the specimen just contain cholesteatoma, (P<0.05),there was a higher level with the specimen just contain cholesteatoma than thegranulation tissue,(P<0.05); The expression of IL-1αin the specimen includingcholesteatoma and granulation tissue have a higher level than the granulationtissue and the specimen just contain cholesteatoma, (P<0.05), there was a higherlevel with the specimen just contain cholesteatoma than the granulation tissuetoo.(P<0.05).
5. There was a positively correlated about the expression of TNF-αand IL-1αin thespecimen including cholesteatoma(r=0.972,P<0.01) ,also in the granulation tissue(r = 0.974, P<0.01); It showed statistically significant correlation betweenexpression of TNF-αand the ability of erosion of middle ear cholesteatoma andgranulation tissue (P<0.01).There was a correlation between the expression ofIL-1αand the ability of erosion of middle ear cholesteatoma and granulation tissue(P<0.01). The lower of the gray value and the higher density of the expression ofTNF-αand IL-α, the stronger of the bone resorption.
Conclutions
1. The TNF-αand IL-1αin the adult middle ear was highly expressed, comparedwith the skin of external auditory of healthy people,explained that there is animportant role in the forming of cholesteatoma and granulation tissue.Theexpression is correlated, suggesting that the interation between them.
2. There was a higher expression of TNF-αand IL-1αin the presence of includingcholesteatoma and granulation tissues than that in the only granulation tissue,indicating that the formation of cholesteatoma and granulation tissue is alwaysin the process of local inflammation.
3. There was the statistically significant correlation between the expression ofTNF-αand IL-1αand the degree of bone resorption, suggesting that TNF-αandIL-1αmay be associated with the bone destruction mechanism of cholesteatomaand granulation tissue.
4. The high expresstion of TNF-αand IL-1αin cholesteatoma and granulation tissue that play an important role in bone destruction ,which for reveal thepathology and molecular mechanism provides a experimental basis.
5. The high expresstion of TNF-αand IL-1αin cholesteatoma and granulationtissue for fully understand their biological behavior provides a theoreticle basis.
方法：选取山西医科大学第一临床医学院耳鼻咽喉-头颈外科行慢性化脓性中耳炎乳突根治手术的81例患者，其中胆脂瘤标本35例，肉芽组织标本20例，包含胆脂瘤和肉芽组织的标本26例，上述病例均为单侧耳发病，详细记录患者的年龄、性别、发病时间、听力损失程度及术中所见骨质破坏程度，选取乳突根治术患者正常外耳道上皮组织10例作为对照，所有标本均经10%中性甲醛固定，常规石蜡包埋，采用免疫组化二步法和计算机图像定量分析系统测定TNF-α和IL-1α分别在上述三型标本中的表达情况。
结果：
1. TNF-α和IL-1α在中耳胆脂瘤中的表达阳性率分别为89%和86%，经χ~2连续性校正检验，与正常外耳道上皮对比差异有统计学意义（P<0.05）。TNF-α在正常外耳道上皮的阳性表达表现为镜下胞浆可见淡棕色颗粒，密度低，位于上皮表层；胆脂瘤中TNF-α阳性表达位于上皮全层，以基底层密度最高，镜下胞浆呈现密集棕黄色颗粒，IL-1α在外耳道上皮中呈弱阳性表达，仅在基底层可见散在的阳性细胞；在胆脂瘤中的阳性表达位于上皮全层，以基底层和棘层密度最高，镜下胞浆呈现高密度的棕黄色颗粒。
2. TNF-α和IL-1α在肉芽组织中的表达阳性率分别为85%和80%，经Fisher确切概率法检验，与正常外耳道上皮对比差异有统计学意义（P<0.05），两者在肉芽组织中的表达大多位于中性粒细胞和巨噬细胞中，且血管内皮细胞显示中等阳性，镜下胞浆可见高密度棕黄色颗粒。
3. TNF-α和IL-1α在包含有胆脂瘤和肉芽组织的标本中阳性表达率分别为96%和88%，镜下可见阳性细胞表达从基底层向角质层逐渐增强，肉芽组织周围和棘细胞层染色较深。
4.经单因素方差分析可得出：包含有胆脂瘤和肉芽的组织TNF-α的表达程度高于仅有肉芽的组织和仅有胆脂瘤的组织（P<0.01），仅有胆脂瘤的组织表达高于仅有肉芽的组织（P<0.01），IL-1α在包含有胆脂瘤和肉芽的组织中表达程度高于仅有肉芽的组织和仅有胆脂瘤组织（P<0.01），在仅有胆脂瘤的组织中的表达高于仅有肉芽的组织（P<0.01）。
5.含有胆脂瘤的组织中TNF-α和IL-1α两者的表达呈正相关(r=0.972，P<0.01)，在肉芽组织中TNF-α和IL-1α的表达亦呈正相关（r=0.974，P<0.01)；胆脂瘤和肉芽组织骨质破坏力与TNF-α和IL-1α的表达有显著相关性(均为P<0.01)。TNF-α和IL-1α表达灰度值越低,表达密度越高,胆脂瘤和肉芽组织的骨质破坏力越强。
结论：
1. TNF-α、IL-1α在成人中耳胆脂瘤和肉芽组织中呈高表达，说明两者在胆脂瘤和肉芽组织形成中起着重要作用，TNF-α和IL-1α的表达有相关性，提示二者的形成相互影响。
2. TNF-α和IL-1α在包含有胆脂瘤和肉芽的组织中表达程度高于仅有肉芽和仅有胆脂瘤的组织，说明胆脂瘤和肉芽组织形成过程中局部始终处于炎症状态。
3.中耳骨质破坏程度与TNF-α和IL-1α的表达有显著相关性，提示TNF-α和IL-1α可能与胆脂瘤和肉芽组织的骨质破坏机制密切相关。
4. TNF-α和IL-1α在胆脂瘤和肉芽组织的高表达与胆脂瘤的骨质破坏及增殖能力具有非常重要的作用，为进一步揭示胆脂瘤的病理分子机制提供了实验基础。
5. TNF-α和IL-1α在胆脂瘤和肉芽组织的异常表达状态，对全面了解胆脂瘤和肉芽组织的生物学行为奠定理论基础。
Object To study the expression of tumor necrosis factor-alpha (TNF-α) andinterleukin -1-alpha(IL-1α) in human middle ear cholesteatoma and granulation tissuefor discussing their roles in the otitis media development.
Methods The specimen are selected from the First Clinical Medical College ofShanxi Medical University, department of Otolaryngology-Head and Neck , including35 cases from cholesteatoma specimens, 20 cases from granulation tissue specimensand 26 cases from both including cholesteatoma and granulation tissuespecimens.Write down their age, gender, illness duration, degree of hearing loss,andobserve their bone damage by surgery meanwhile.In addition, patients with normalexternal ear canal epithelium in 10 cases as controls. Finally,all specimens were fixedin 10% neutral formalin and paraffin-embedded.To measure the expression of TNF-αand IL-1αin the middle ear cholesteatoma and granulation tissue byimmunohistochemical technology and computer image analysis system.
Results
1. The percent of positive expression of TNF-αand IL-1αin middle earcholesteatoma were 89% and 86%, as compared with the normal ear canal skin,there were significant differences in expression (P<0.05.). The positive expressionof TNF-αin normal external ear canal epithelium cytoplasm showed a light browngranular and lowly density,which have a higher dendity expression in allcholesteatoma epithelial layers, the highest density in the basal layer, subepithelialconnective tissue is also have a positive expression.There is a weakly expressionof IL-1αin the external auditory canal epithelium and the positive cells onlyscattered in the basal layer,which have a higher expression in the wholecholesteatoma layer, with the highest density in basal and spinous layer.
2. The percent of positive expression of TNF-α, and IL-1αin granulation tissue were85% and 80%, as compared with the normal ear canal skin, there were significantdifferences in expression (P<0.05.).Both have a higher expression in neutrophilegranulocyte and macrophages,also in the vascular endothelial cells.
3. The percent of positive expression of TNF-αand IL-1αin the specimen includingcholesteatoma and granulation tissue were 96% and 88%,by the microscope we could observed that positive cells gradually increased from the basal layer to thestratum corneum,particularly have a higher expression around the granulationtissue and stratum spinosum epidermidis.
4. By the Oneway ANOVA analysis can observed :The expression of TNF-αin thespecimen including cholesteatoma and granulation tissue have a higher level thanthe granulation tissue and the specimen just contain cholesteatoma, (P<0.05),there was a higher level with the specimen just contain cholesteatoma than thegranulation tissue,(P<0.05); The expression of IL-1αin the specimen includingcholesteatoma and granulation tissue have a higher level than the granulationtissue and the specimen just contain cholesteatoma, (P<0.05), there was a higherlevel with the specimen just contain cholesteatoma than the granulation tissuetoo.(P<0.05).
5. There was a positively correlated about the expression of TNF-αand IL-1αin thespecimen including cholesteatoma(r=0.972,P<0.01) ,also in the granulation tissue(r = 0.974, P<0.01); It showed statistically significant correlation betweenexpression of TNF-αand the ability of erosion of middle ear cholesteatoma andgranulation tissue (P<0.01).There was a correlation between the expression ofIL-1αand the ability of erosion of middle ear cholesteatoma and granulation tissue(P<0.01). The lower of the gray value and the higher density of the expression ofTNF-αand IL-α, the stronger of the bone resorption.
Conclutions
1. The TNF-αand IL-1αin the adult middle ear was highly expressed, comparedwith the skin of external auditory of healthy people,explained that there is animportant role in the forming of cholesteatoma and granulation tissue.Theexpression is correlated, suggesting that the interation between them.
2. There was a higher expression of TNF-αand IL-1αin the presence of includingcholesteatoma and granulation tissues than that in the only granulation tissue,indicating that the formation of cholesteatoma and granulation tissue is alwaysin the process of local inflammation.
3. There was the statistically significant correlation between the expression ofTNF-αand IL-1αand the degree of bone resorption, suggesting that TNF-αandIL-1αmay be associated with the bone destruction mechanism of cholesteatomaand granulation tissue.
4. The high expresstion of TNF-αand IL-1αin cholesteatoma and granulation tissue that play an important role in bone destruction ,which for reveal thepathology and molecular mechanism provides a experimental basis.
5. The high expresstion of TNF-αand IL-1αin cholesteatoma and granulationtissue for fully understand their biological behavior provides a theoreticle basis.
引文
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[1]王辉兵,徐志文,唐安洲,等.中耳胆脂瘤基质金属蛋白酶2, 9的活性检测及临床意义[J].临床耳鼻咽喉科杂志,2004,18(10):620-622.
[2]温文胜,黄光武,余奇松,等.胆脂瘤中纤溶酶原激活物质测定[J].耳鼻咽喉-头颈外科杂志, 2001,8(4):241-244.
[3]王辉兵,徐志文,唐安洲.胆脂瘤型中耳炎骨质破坏机制的组织酶学研究[J].耳鼻咽喉-头颈外科杂志,2002,9(1):62-64.
[4]Yellon RF，Leonrd G,Marucha PT,et al.Characterization of cytokines present inmiddle ear effusions[J].Laryngoscope,1991;101(2):165-169.
[5]Yellon RF，Leonrd G,Marucha PT,et al.Demonstration of interleukin 6 in middleear effusions[J].Arch Otolaryngol Head Neck Surg,1992;118(7):745-748.
[6]张全安，梁建民.渗出性中耳炎渗出液的病理转化和肉芽组织形成[J].临床耳鼻咽喉科杂志，1999,13(1):8-11.
[7]孙斌，陈钢，许珉，张全安.肿瘤坏死因子和慢性中耳炎的关系[N]，西安交通大学学报，2004,10（5）.
[8]沈玲，葛文胜，王领台.肿瘤坏死因子-a在中耳胆脂瘤中的定量表达及其在骨吸收中的作用[J]。中国医药导报，2008,3（9），34-36.
[9]李正贤,李学佩,刘庚勋,等.肿瘤坏死因子-α在中耳胆脂瘤中的表达及其意义[J].耳鼻咽喉-头颈外科杂志,2000,7(5):287-289.
[10]Akimoto R ,Pawankar R ,Yagi T,et al.Acquired and congenitalcholesteatoma :determination of tumor necrosis factor2alpha ,intercellularadhesion molecule21 ,interleukin212alpha and lymphocyte functional antigen21in the inflammatory process [ In Process Citation] [J ]1ORL J OtorhinolaryngolRelat Spec ,2000 ;62 (5) :257-651.
[11]Sastry KV ,Sharma SC ,Mann SB ,et al . Aural cholesteatoma :role of tumornecrosis factor2alpha in bone destruction[J ] . AmJ Otol ,1999 ;20 (2) :158-160.
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[15]KinoshitaK. The role of interleuk in- 1 alpha, tumor necrosis factor- alpha andparathyro idho rmone- related protein in bone resorption of cholesteatoma otitis[J].Nippon - J ibiinkoka - Gakkai- Kaiho, 1994 ;Aug 97 (8):147- 148.
[16]YanSD, Huang CC. Tumor necrosis factor alpha in middle ear cholesteatoma andits effect on keratinocytes in vitro[ J ]. Ann Otol Rhinol Laryngol, 1996;100 (2) :157-161.
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[18]Chodynicki S, Soroczynska J. TNF-alpha in serum of patients withcholesteatoma[J]. Otolaryngo l Po l, 1994;48 (3) : 279-281.
[19]Kuczkowski J, Sakowicz-Burkiewicz M, I ycka- wieszewska E, Mikaszewski B,Pawe czyk T,Expression of Tumor Necrosis Factor-a, Interleukin-1a,Interleukin-6 and Interleukin-10 in Chronic Otitis Media with BoneOsteolysis.ORL J Otorhinolaryngol Relat Spec.2011;73(2):93-9.Epub 2011 Feb10.
[20]沈玲，葛文胜，王领台.肿瘤坏死因子-α在中耳胆脂瘤中的定量表达及其在骨吸收中的作用[J]。中国医药导报，2008,3（9），34-36.
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[24]Yetiser S,Satar B,Aydin N. Expression of epidermal growth factor, tumornecrosis factor-alpha, and interleukin-1alpha in chronic otitis media with orwithout cholesteatoma[J].Otology & neurotology,2010;105:220~227.
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[28]Tanaka Y,Kojima H,Miyazaki H,et al.Roles of cytokines and cell cycle regulatingsubstances in proliferation of cholesteatomaepithelium.Laryngoscope,1999;109:1102-1107.
[2]Sastry KV ,Sharma SC ,Mann SB ,et al .Αural cholesteatoma :role of tumornecrosis factor2alpha in bone destruction[J ] .Αm J Otol ,1999 ;20 (2) :158-160.
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[1]王辉兵,徐志文,唐安洲,等.中耳胆脂瘤基质金属蛋白酶2, 9的活性检测及临床意义[J].临床耳鼻咽喉科杂志,2004,18(10):620-622.
[2]温文胜,黄光武,余奇松,等.胆脂瘤中纤溶酶原激活物质测定[J].耳鼻咽喉-头颈外科杂志, 2001,8(4):241-244.
[3]王辉兵,徐志文,唐安洲.胆脂瘤型中耳炎骨质破坏机制的组织酶学研究[J].耳鼻咽喉-头颈外科杂志,2002,9(1):62-64.
[4]Yellon RF，Leonrd G,Marucha PT,et al.Characterization of cytokines present inmiddle ear effusions[J].Laryngoscope,1991;101(2):165-169.
[5]Yellon RF，Leonrd G,Marucha PT,et al.Demonstration of interleukin 6 in middleear effusions[J].Arch Otolaryngol Head Neck Surg,1992;118(7):745-748.
[6]张全安，梁建民.渗出性中耳炎渗出液的病理转化和肉芽组织形成[J].临床耳鼻咽喉科杂志，1999,13(1):8-11.
[7]孙斌，陈钢，许珉，张全安.肿瘤坏死因子和慢性中耳炎的关系[N]，西安交通大学学报，2004,10（5）.
[8]沈玲，葛文胜，王领台.肿瘤坏死因子-a在中耳胆脂瘤中的定量表达及其在骨吸收中的作用[J]。中国医药导报，2008,3（9），34-36.
[9]李正贤,李学佩,刘庚勋,等.肿瘤坏死因子-α在中耳胆脂瘤中的表达及其意义[J].耳鼻咽喉-头颈外科杂志,2000,7(5):287-289.
[10]Akimoto R ,Pawankar R ,Yagi T,et al.Acquired and congenitalcholesteatoma :determination of tumor necrosis factor2alpha ,intercellularadhesion molecule21 ,interleukin212alpha and lymphocyte functional antigen21in the inflammatory process [ In Process Citation] [J ]1ORL J OtorhinolaryngolRelat Spec ,2000 ;62 (5) :257-651.
[11]Sastry KV ,Sharma SC ,Mann SB ,et al . Aural cholesteatoma :role of tumornecrosis factor2alpha in bone destruction[J ] . AmJ Otol ,1999 ;20 (2) :158-160.
[12]Amar MS ,Wishahi HF ,Zakhary MM. Clinical and biochemical studies of bonedestruction in cholesteatoma[J ] . Laryngol Otol ,1996 ;110 (6) :534-5391.
[13]Marenda SA ,Aufdemorte TB. Localization of cytokines in cholesteatomatissue[J ] . Otolaryngol Head Neck Surg ,1995;112 (3) :359-368.
[14]Iino Y, To riyamaM , Ogawa H, et al. Cho lesteatoma debris as an activato r ofhuman monocytes[J]. A cta O to laryngo l (Stockh) , 1990;110: 410-415.
[15]KinoshitaK. The role of interleuk in- 1 alpha, tumor necrosis factor- alpha andparathyro idho rmone- related protein in bone resorption of cholesteatoma otitis[J].Nippon - J ibiinkoka - Gakkai- Kaiho, 1994 ;Aug 97 (8):147- 148.
[16]YanSD, Huang CC. Tumor necrosis factor alpha in middle ear cholesteatoma andits effect on keratinocytes in vitro[ J ]. Ann Otol Rhinol Laryngol, 1996;100 (2) :157-161.
[17]KinoshitaK. The role of interleukin- 1 alpha, tumor necrosis factor- alpha andparathyroid hormone- related protein in bone resorption of cholesteatoma otitis[J].Nippon - J ibiinkoka - Gakkai- Kaiho, 1994; Aug 97 (8):147- 148.
[18]Chodynicki S, Soroczynska J. TNF-alpha in serum of patients withcholesteatoma[J]. Otolaryngo l Po l, 1994;48 (3) : 279-281.
[19]Kuczkowski J, Sakowicz-Burkiewicz M, I ycka- wieszewska E, Mikaszewski B,Pawe czyk T,Expression of Tumor Necrosis Factor-a, Interleukin-1a,Interleukin-6 and Interleukin-10 in Chronic Otitis Media with BoneOsteolysis.ORL J Otorhinolaryngol Relat Spec.2011;73(2):93-9.Epub 2011 Feb10.
[20]沈玲，葛文胜，王领台.肿瘤坏死因子-α在中耳胆脂瘤中的定量表达及其在骨吸收中的作用[J]。中国医药导报，2008,3（9），34-36.
[21]Schilling V,Negri B,Bujia J,et al.Possible role of interleukin-1 alpha andinterleukin 1 beta in the pathogenesis of cholesteatoma of the middle ear.Am JOtol,1992;13:350-355.
[22]Bujia J,Kim C,Boyle D,et al.Quantitative analysis of interleukin-1-alpha geneexpression in middle ear cholesteatoma[J]. Laryngoscope,1996,106(2 Pt1):217-220.
[23]Kim CS,Lee CH,Chung JW,et al.Interleukin-1 alpha, Interleukin-1 beta andInterleukin-8 gene expression in human aural cholesteatomas[J].ActaOtolaryngol,1996,116(2):302-306.
[24]Yetiser S,Satar B,Aydin N. Expression of epidermal growth factor, tumornecrosis factor-alpha, and interleukin-1alpha in chronic otitis media with orwithout cholesteatoma[J].Otology & neurotology,2010;105:220~227.
[25]Mundy GR et al.Role of cytokines in bone resorption[J].CellBiochem,1993;53:296~300.
[26]Sastry KV ,Sharma SC ,Mann SB ,et al . Aural cholesteatoma :role of tumornecrosis factor -alpha in bone destruction[J ] . AmJ Otol ,1999 ;20 (2) :158-160.
[27]Bujia J.Interleukin-1(IL-1) and IL-1-receptor antagonist(IL-1-RA) in middle earcholesteatoma:a analysis of protein production and biological activity[J].Eur ArchOtorhino laryngol,1996;253(4-5):252-255.
[28]Tanaka Y,Kojima H,Miyazaki H,et al.Roles of cytokines and cell cycle regulatingsubstances in proliferation of cholesteatomaepithelium.Laryngoscope,1999;109:1102-1107.