中耳胆脂瘤组织LPS、VDR、MMP-9的检测及相关性分析
|
摘要
目的:探讨脂多糖、维生素D受体、基质金属蛋白酶-9蛋白在中耳胆脂瘤中的含量及表达的情况和它们之间的关系。
方法:标本来自在广西医科大学第一附属医院耳鼻咽喉科行手术治疗的29例胆脂瘤型中耳炎患者,其中18例来自这些胆脂瘤患者的正常外耳道皮肤作为对照组。所有胆脂瘤病例全部经病理确诊。标本分两部分收集,一部分用无热原冻存管于-80℃下保存用作LPS含量测定,采用鲎变性细胞溶解物测定法(LAL法);另一部分经甲醛固定、石蜡包埋,用作鼠抗人VDR与鼠抗人MMP-9蛋白的免疫组化,检测其表达情况。
结果:结果发现,LPS在胆脂瘤组织中含量比在正常外耳道皮肤组织中表达高(均为取对数后数值):胆脂瘤组中位数(Median,M)=0.7390,四分位数间距(interquartile range,IQR)=0.6203;外耳道组M=-0.2538,IQR=1.1692(P＜0.05)。在两种类型的胆脂瘤组织中,广泛型组M=0.8403,IQR=0.5254;局限型组M=0.4048,IQR=0.6139(P＜0.05),广泛型的LPS含量比局限型的高。VDR蛋白在外耳道上皮呈阳性表达,而在胆脂瘤上皮呈中、低表达。VDR蛋白阳性定位于细胞核,呈棕黄色至褐色颗粒状。在胆脂瘤组,VDR阳性率为93.1%,与外耳道皮肤相比,具有显著差异(P＜0.05)。MMP-9蛋白在胆脂瘤上皮中基本呈阳性表达,而在外耳道皮肤上皮中呈中、低表达或不表达。MMP-9蛋白阳性定位于细胞质,呈棕黄色颗粒状。在胆脂瘤标本中,MMP-9阳性细胞率为79.3%,与外耳道皮肤相比,具有显著差异(P＜0.05);在两种类型的胆脂瘤中,MMP-9和VDR的表达差异无统计学意义(P＞0.05)。经Spearman检验LPS、VDR、MMP-9之间无相关关系。
结论:
1、LPS在胆脂瘤中的含量高于外耳道皮肤,且胆脂瘤广泛型组比局限型组含量增高,提示LPS与胆脂瘤的形成及其骨质破坏有关。
2、VDR在外耳道皮肤的表达高于胆脂瘤上皮,提示VDR与胆脂瘤的发生有关。
3、MMP-9在胆脂瘤上皮中的表达高于外耳道皮肤,提示MMP-9与胆脂瘤的发生有关。
4、LPS、VDR、MMP-9三者无相关关系,提示三者在中耳胆脂瘤发生中的作用尚无明确的联系。
Objective: To investigate the expression and concentration of lipopolysaccharide, vitamin D receptor and matrix metalloproteinase-9 in middle ear cholesteatoma and discuse their relations.
Methods: 29 cases of middle ear cholesteatoma tissue we studied were come from the patients who were operated in First Affliliated Hospital of Guangxi Medical University, 18 cases of external auditory canal tissue were acquired from those patients at the same time, and all of them had been verified by pathology. Tissues were stored into 2 parts, ones were stored without any additive in pyrogen-free tubes at -80 C until further processing, the other ones were formalin-fixed, paraffin-embedded for immunohistochemical test. The concentrations of LPS were detected by limulus amebocyte lysate assay (LAL-assay) , and expression of VDR and MMP-9 protein in formalin-fixed, paraffin-embedded tissues were detected by immunohistochemical method.
Results: The result showed that the concentrations of LPS in cholesteatoma are higher than in external auditory canal tissues (the numbers are logarithmed): Group of Cholesteatoma: M=0. 7390, IQR=0. 6203; Group of external auditory canal tissues : M=-0.2538 , IQR=1. 1692 (P<0.05) . Between the two-type groups: Group of intensive type: M=0. 8403, IQR=0. 5254; Group of localized type: M=0. 4048, IQR=0. 6139, the concentrations of LPS were higher in extensive cholesteatoma in comparison with localized cholesteatoma (P<0.05) . VDR was located at karyon, taking on yellow brown pellet, could be detected in all external auditory canal epithelium but there were less positive cells in middle ear cholesteatoma. In cholesteatoma epithelium samples, VDR were 93.1%. MMP-9 was located at cytoplasmic, taking on yellow brown pellet, could be detected in almost middle ear cholesteatoma epithelium but there were less positive cells in external auditory canal epithelium. In cholesteatoma epithelium samples, MMP-9 were 79.3%. Compared with external auditory canal epithelium, the expression of MMP-9 was higher in middle ear cholesteatoma epithelium (P<0.05), but was lower in expression of VDR(P<0.05). They didn't have significant difference in two types of cholesteatoma epithelium(P>0.05). LPS, MMP-9 and VDR weren't significant correlated by Spearman test.
Conclusion:
1 The concentrations of LPS were higher in cholesteatoma compared with external auditory canal, and were higher in extensive types compared with localized ones, which hints LPS was responsible for middle ear cholesteatoma and the bone erosion of cholesteatoma.
2 The expression of VDR was higher in external auditory canal compared with cholesteatoma, which hints VDR was responsible for middle ear cholesteatoma.
3 The expression of MMP-9 was higher in cholesteatoma compared with external auditory canal, which hints MMP-9 was responsible for middle ear cholesteatoma.
4 There's no correlation among LPS, VDR and MMP-9, which hints the relationships among the three were unkown.
方法:标本来自在广西医科大学第一附属医院耳鼻咽喉科行手术治疗的29例胆脂瘤型中耳炎患者,其中18例来自这些胆脂瘤患者的正常外耳道皮肤作为对照组。所有胆脂瘤病例全部经病理确诊。标本分两部分收集,一部分用无热原冻存管于-80℃下保存用作LPS含量测定,采用鲎变性细胞溶解物测定法(LAL法);另一部分经甲醛固定、石蜡包埋,用作鼠抗人VDR与鼠抗人MMP-9蛋白的免疫组化,检测其表达情况。
结果:结果发现,LPS在胆脂瘤组织中含量比在正常外耳道皮肤组织中表达高(均为取对数后数值):胆脂瘤组中位数(Median,M)=0.7390,四分位数间距(interquartile range,IQR)=0.6203;外耳道组M=-0.2538,IQR=1.1692(P＜0.05)。在两种类型的胆脂瘤组织中,广泛型组M=0.8403,IQR=0.5254;局限型组M=0.4048,IQR=0.6139(P＜0.05),广泛型的LPS含量比局限型的高。VDR蛋白在外耳道上皮呈阳性表达,而在胆脂瘤上皮呈中、低表达。VDR蛋白阳性定位于细胞核,呈棕黄色至褐色颗粒状。在胆脂瘤组,VDR阳性率为93.1%,与外耳道皮肤相比,具有显著差异(P＜0.05)。MMP-9蛋白在胆脂瘤上皮中基本呈阳性表达,而在外耳道皮肤上皮中呈中、低表达或不表达。MMP-9蛋白阳性定位于细胞质,呈棕黄色颗粒状。在胆脂瘤标本中,MMP-9阳性细胞率为79.3%,与外耳道皮肤相比,具有显著差异(P＜0.05);在两种类型的胆脂瘤中,MMP-9和VDR的表达差异无统计学意义(P＞0.05)。经Spearman检验LPS、VDR、MMP-9之间无相关关系。
结论:
1、LPS在胆脂瘤中的含量高于外耳道皮肤,且胆脂瘤广泛型组比局限型组含量增高,提示LPS与胆脂瘤的形成及其骨质破坏有关。
2、VDR在外耳道皮肤的表达高于胆脂瘤上皮,提示VDR与胆脂瘤的发生有关。
3、MMP-9在胆脂瘤上皮中的表达高于外耳道皮肤,提示MMP-9与胆脂瘤的发生有关。
4、LPS、VDR、MMP-9三者无相关关系,提示三者在中耳胆脂瘤发生中的作用尚无明确的联系。
Objective: To investigate the expression and concentration of lipopolysaccharide, vitamin D receptor and matrix metalloproteinase-9 in middle ear cholesteatoma and discuse their relations.
Methods: 29 cases of middle ear cholesteatoma tissue we studied were come from the patients who were operated in First Affliliated Hospital of Guangxi Medical University, 18 cases of external auditory canal tissue were acquired from those patients at the same time, and all of them had been verified by pathology. Tissues were stored into 2 parts, ones were stored without any additive in pyrogen-free tubes at -80 C until further processing, the other ones were formalin-fixed, paraffin-embedded for immunohistochemical test. The concentrations of LPS were detected by limulus amebocyte lysate assay (LAL-assay) , and expression of VDR and MMP-9 protein in formalin-fixed, paraffin-embedded tissues were detected by immunohistochemical method.
Results: The result showed that the concentrations of LPS in cholesteatoma are higher than in external auditory canal tissues (the numbers are logarithmed): Group of Cholesteatoma: M=0. 7390, IQR=0. 6203; Group of external auditory canal tissues : M=-0.2538 , IQR=1. 1692 (P<0.05) . Between the two-type groups: Group of intensive type: M=0. 8403, IQR=0. 5254; Group of localized type: M=0. 4048, IQR=0. 6139, the concentrations of LPS were higher in extensive cholesteatoma in comparison with localized cholesteatoma (P<0.05) . VDR was located at karyon, taking on yellow brown pellet, could be detected in all external auditory canal epithelium but there were less positive cells in middle ear cholesteatoma. In cholesteatoma epithelium samples, VDR were 93.1%. MMP-9 was located at cytoplasmic, taking on yellow brown pellet, could be detected in almost middle ear cholesteatoma epithelium but there were less positive cells in external auditory canal epithelium. In cholesteatoma epithelium samples, MMP-9 were 79.3%. Compared with external auditory canal epithelium, the expression of MMP-9 was higher in middle ear cholesteatoma epithelium (P<0.05), but was lower in expression of VDR(P<0.05). They didn't have significant difference in two types of cholesteatoma epithelium(P>0.05). LPS, MMP-9 and VDR weren't significant correlated by Spearman test.
Conclusion:
1 The concentrations of LPS were higher in cholesteatoma compared with external auditory canal, and were higher in extensive types compared with localized ones, which hints LPS was responsible for middle ear cholesteatoma and the bone erosion of cholesteatoma.
2 The expression of VDR was higher in external auditory canal compared with cholesteatoma, which hints VDR was responsible for middle ear cholesteatoma.
3 The expression of MMP-9 was higher in cholesteatoma compared with external auditory canal, which hints MMP-9 was responsible for middle ear cholesteatoma.
4 There's no correlation among LPS, VDR and MMP-9, which hints the relationships among the three were unkown.
引文
1 Diks SH,RichelDJ,Peppelenbosch ME LPS signal transduction:the picture is becoming more complex.Curr Top Med Chem,2004,4(11):1115-1126.
2 Kidachi Y,Yamaguchi H,Umetsu H,etc.Interferon-gamma and lipopolysaccharide stimulation increases matrix metalloproteinase-9expression and enhances invasion activity in ras/myc-transformed serum-free mouse embryo cells.Cell Biol Int.2007,31(12):1511-1517.
3 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.临床耳鼻咽喉科杂志.2004,10(18):620-622
4 Kobayashi H,Asano K,Ken-ichi Kanai,etc.Suppressive activity of vitamin D_3 on matrix metalloproteinase production from cholesteatoma keratinocytes in vitro.Mediators of Inflammation.2005,4:210-215.
5 孙文忠,徐志文,唐安洲,等.细胞周期调控因子在中耳胆脂瘤上皮中的表达.中国耳鼻咽喉头颈外科.2005,2(12):95-97.
6 胡宗涛.VDR在乳腺癌中的表达的研究.安徽医科大学硕士论文.
7 祝威,谢燕,王苹等.两种基质金属蛋白酶在胆脂瘤和中耳癌中的表达.中华耳鼻咽喉科杂志.2001,36(2):119-122.
8 Bannerman DD,Paape MJ,Hare WR,etc.Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge.J Dairy Sci.2003,86(10):3128-3137.
9 Vesy CJ,Kitchens RL,Wolfbauer G,etc.Lipopolysaccharide-binding protein and phospholipid transfer protein release lipopolysaccharides from gram-negative bacterial membranes.2000,68(5):2410-2417.
10 Jerala R.Structural biology of the LPS recognition.Int J Med Microbiol.2007,297(5):353-363.
11 Kitchens RL,Thompson PA.Modulatory effects of sCD14 and LBP on LPS-host cell interactions.J Endotoxin Res.2005,11(4):225-229.
12 Li CH,Wang JH,Redmond HP.Bacterial lipoprotein-induced self-tolerance and cross-tolerance to LPS are associated with reduced IRAK-1 expression and MyD88-IRAK complex formation.J Leukoc Biol.2006,79(4):867-875.
13 Medvedev AE,Piao W,Shoenfelt J,etc.Role of TLR4 tyrosine phosphorylation in signal transduction and endotoxin tolerance.J Biol Chem.2007 282(22):16042-16053.
14 许昱,陶泽璋,华清泉,等.中耳胆脂瘤中核因子-κK B的表达与活化.中华耳鼻咽喉头颈外科杂志.2006,41(6):455-459.
15 Lazarov S,Balutsov M,Ianev E.The role of bacterial endotoxins,receptors and cytokines in the pathogenesis of septic(endotoxin)shock Vutr Boles.2000,32(4):33-40.
16 Cohen J.The detection and interpretation of endotoxaemia.Intensive Care Med.2000,26(1):51-56.
17 Albino AP,Kimmelman CE Parisier SC.Cholesteatoma:a molecular and cellular puzzle.Am J Otol.1998,19(1):7-19.
18 石磊,张建中,王荣光.胆脂瘤型中耳炎细菌检测.中国耳鼻咽喉.头颈外 科.2004,11(6):366.
19 Vorobev AA, Borisova EV, Borisov VA. Gram-negative virulent bacterial lipopolysaccharide: role in infection and immunity. Vestn Ross Akad Med Nauk.1997, 3:10-13.
20 Peek FA, Huisman MA, Berckmans RJ, etc. Lipopolysaccharide concentration and bone resorption in cholesteatoma. Otol Neurotol. 2003 Sep;24(5):709-13.
21 Dumitrescu AL, Abd-El-Aleem S, Morales-Aza B, etc.A model of periodontitis in the rat: effect of lipopolysaccharide on bone resorption, osteoclast activity, and local peptidergic innervation. J Clin Periodontol. 2004,31(8):596-603.
22 Rogers JE, Li F, Coatney DD,etc.Actinobacillus actinomycetemcomitans lipopolysaccharide-mediated experimental bone loss model for aggressive periodontitis. J Periodontol. 2007 ,78(3):550-558.
23 Deluca HF, Zierold C. Mechnisms and functions of virmain D.Nurt Rev, 1998:56(22):4-10.
24 Carlbeg C, Polly P Gene regulation by vitamin D3. Crit Re Ekuyarot Gene ExPr.l998:8(1):19-42.
25 Holick MF. Vitamin D: A millenium perspective. J Cell Biochem. 2003,88:296-307.
26 Zittermann A. Vitamin D in preventive medicine: are we ignoring the evidence?. Br J Nutr, 2003,89:552-572.
27 Haussler MR, Whitfield GK, Haussler CA, et al. The nuclear vitamin D receptor: biological and molecular regulatory properties revealed. J Bone Miner Res, 1998,13:325-349.
28 Sun J, Kong J, Duan Y, etc. Increased NF-kappaB activity in fibroblasts lacking the vitamin D receptor. Am J Physiol Endocrinol Metab. 2006, 291(2): 315-322.
29 Holick MR Vitamin D: A millenium perspective. J Cell Biochem. 2003,88(2):296-307.
30 Parish JL. Topical vitamin D3 analogues: unapproved uses, dosages, or indications. Clin Dermatol, 2002,20:558-562.
31 Ermis O, Alpsoy E, Cetin L, etc. Is the efficacy of psoralen plus ultraviolet A therapy for vitiligo enhanced by concurrent topical calcipotriol? A placebo-controlled double-blind study. Br J Dermatol, 2001,145:472-5.
32 Inoue M, Matsui T, Nishibu A, etc. Regulatory effectsof l,alpha 25-dihydroxyvitamin D3 on inflammatory responses in psoriasis. Eur J Dermatol, 1998,8:16-20.
33 Pramanik R, Asplin JR, Lindeman C, etc. Lipopolysaccharide negatively modulates vitamin D action by down-regulating expression of vitamin D-induced VDR in human monocytic THP-1 cells. Cell Immunol. 2004,232(1-2): 137-143
34 Timms PM,Mannan N,Hitman GA.Circulating MMP9, vitamin D and variation in the TIMP-1 response with VDR genotype: mechanisms for inflammatory damage in chronic disorders? QJM. 2002,95(12):787-796.
35 Kleiner DE,Stetler-Stevenson WG.Matrix metalloproteinases and metastasis.Cancer Chemother Pharmacol.1999,43:42-51.
36 Wojtowicz-Praga SM,Dickson RB,Hawkins MJ.Matrix metalloproteinase inhibitors.Invest New Drugs.1997,15(1):61-75.
37 Dallari S,Cavani A,Bergamini G etc.Integrin expression in middle ear cholesteatoma.Acta Otolaryngol.1994,114(2):188-192.
38 Moriyama H,Honda Y,Huang CC etc.Bone resorption in cholesteatoma:epithelial-mesenchymal cell interaction and collagenase production.Laryngoscope.1987,97:854-859.
39 Sudhoff H,Bujia J,Borkowski G,etc.Basement membrane in middle ear cholesteatoma immunohistochemical and ultrastructural observations.Ann Otol Rhinol Laryngol.1996,105:804-810.
1 Macri JR,Chole RA.Bone erosion in experimental cholesteatoma--the effects of implanted barriers[J].Otolaryngol Head Neck Surg,1985,93(1):3-17.
2 Chole RA,McGinn MD,Tinling SP.Pressure-induced bone resorption in the middle ear.Ann Otol Rhinol Laryngol.1985 Mar-Apr;94(2 Pt 1):165-70.PMID:3994236.
3 Burger EH,Klein-Nulen J.Responses of bone cells to biomechanical forces in vitro[J].Adv Dent Res,1999,13(1):93-98.
4 Park HJ,Park K.Expression of Fas/APO-1 and apoptosis of keratinocytes in human cholesteatoma[J].Laryngoscope,1999,109(4):613-616.
5 Park K,Chun YM,Lee DH.Expression of phospholipase C-gamma1 in experimental cholesteatoma using Mongolian gerbils[J].Acta Otolaryngol,2001,121(4):477-480.
6 孙文忠,徐志文,唐安洲,等.PTKs、CDK4及p15在中耳胆脂瘤上皮的表达[J].临床耳鼻咽喉科杂志,2004,18(10):616-619.
7 Suzuki C,Ohtani I.Bone destruction resulting from rupture of a cholesteatoma sac:temporal bone pathology[J].Otol Neurotol,2004,25(5):674-677.
8 Takahashi N.Bone and bone related biochemical examinations.Bone and collagen related metabolites.Regulatory mechanisms of osteoclast differentiation and function[J].Clin Calcium,2006,16(6):940-947.
9 Wada T,Nakashima T,Hiroshi N,et al.RANKL-RANK signaling in osteoclastogenesis and bone disease[J].Trends Mol Med,2006,12(1):17-25.
10 Lin JM,Callon KE,Lin CQ,et al.Alteration of bone cell function by RANKL and OPG in different in vitro models[J].Eur J Clin Invest,2007, 37(5): 407-415.
11 Uno Y, Saito R. Bone resorption in human cholesteatoma: morphological study with scanning electron microscopy[J].Ann Otol Rhinol Laryngol, 1995, 104(6) :463-468.
12 Jeong JH, Park CW, Tae K. Expression of RANKL and OPG in middle ear cholesteatoma tissue[J]. Laryngoscope,2006,116(7):1180-1184.
13 Lee ZH, Lee SE, Kim CW,et al. IL-1alpha stimulation of osteoclast survival through the PI 3-kinase/Akt and ERK pathways[J]. J Biochem (Tokyo), 2002, 131(1): 161-166.
14 Bujia J, Kim C, Ostos P, et al. Role of interleukin 6 in epithelial hyperproliferation and bone resorption in middle ear cholesteatomas[J]. Eur Arch Otorhinolaryngol, 1996, 253(3): 152-157.
15 Mihara M, Nishimoto N. Interleukin-6 in pathogenesis of rheumatoid arthritis[J].Nippon Rinsho,2005,63(1):158-162.
16 Kitaura H, Zhou P,Kim HJ,et al.M-CSF mediates TNF-induced inflammatory osteolysis[J].J Clin Invest,2005,115(12):3418-3427.
17 Zhang YH,Heulsmann A,Tondravi MM,et al.Tumor necrosis factor-alpha (TNF) stimulates RANKL-induced osteoclastogenesis via coupling of TNF type 1 receptor and RANK signaling pathways[J].J Biol Chem,2001,276(1):563-568.
18 Hotokezaka H,Sakai E, Ohara N,et al. Molecular analysis of RANKL-independent cell fusion of osteoclast-like cells induced by TNF-alpha,lipopolysaccharide,or peptidoglycan[J]. J Cell Biochem, 2007, 101(1): 122-134.
19 Chole RA.Osteoclasts in chronic otitis media,cholesteatoma,and otosclerosis[J].Ann Otol Rhiool Laryngol,1988,97(6):661-666.
20 Abramson M.Collagenase in the mechanism of action of cholesteatoma[J].Ann Otol Rhinol Laryngol,1971,80(3):414.
21 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.
22 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,2001,8(4):241-244.
2 Kidachi Y,Yamaguchi H,Umetsu H,etc.Interferon-gamma and lipopolysaccharide stimulation increases matrix metalloproteinase-9expression and enhances invasion activity in ras/myc-transformed serum-free mouse embryo cells.Cell Biol Int.2007,31(12):1511-1517.
3 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.临床耳鼻咽喉科杂志.2004,10(18):620-622
4 Kobayashi H,Asano K,Ken-ichi Kanai,etc.Suppressive activity of vitamin D_3 on matrix metalloproteinase production from cholesteatoma keratinocytes in vitro.Mediators of Inflammation.2005,4:210-215.
5 孙文忠,徐志文,唐安洲,等.细胞周期调控因子在中耳胆脂瘤上皮中的表达.中国耳鼻咽喉头颈外科.2005,2(12):95-97.
6 胡宗涛.VDR在乳腺癌中的表达的研究.安徽医科大学硕士论文.
7 祝威,谢燕,王苹等.两种基质金属蛋白酶在胆脂瘤和中耳癌中的表达.中华耳鼻咽喉科杂志.2001,36(2):119-122.
8 Bannerman DD,Paape MJ,Hare WR,etc.Increased levels of LPS-binding protein in bovine blood and milk following bacterial lipopolysaccharide challenge.J Dairy Sci.2003,86(10):3128-3137.
9 Vesy CJ,Kitchens RL,Wolfbauer G,etc.Lipopolysaccharide-binding protein and phospholipid transfer protein release lipopolysaccharides from gram-negative bacterial membranes.2000,68(5):2410-2417.
10 Jerala R.Structural biology of the LPS recognition.Int J Med Microbiol.2007,297(5):353-363.
11 Kitchens RL,Thompson PA.Modulatory effects of sCD14 and LBP on LPS-host cell interactions.J Endotoxin Res.2005,11(4):225-229.
12 Li CH,Wang JH,Redmond HP.Bacterial lipoprotein-induced self-tolerance and cross-tolerance to LPS are associated with reduced IRAK-1 expression and MyD88-IRAK complex formation.J Leukoc Biol.2006,79(4):867-875.
13 Medvedev AE,Piao W,Shoenfelt J,etc.Role of TLR4 tyrosine phosphorylation in signal transduction and endotoxin tolerance.J Biol Chem.2007 282(22):16042-16053.
14 许昱,陶泽璋,华清泉,等.中耳胆脂瘤中核因子-κK B的表达与活化.中华耳鼻咽喉头颈外科杂志.2006,41(6):455-459.
15 Lazarov S,Balutsov M,Ianev E.The role of bacterial endotoxins,receptors and cytokines in the pathogenesis of septic(endotoxin)shock Vutr Boles.2000,32(4):33-40.
16 Cohen J.The detection and interpretation of endotoxaemia.Intensive Care Med.2000,26(1):51-56.
17 Albino AP,Kimmelman CE Parisier SC.Cholesteatoma:a molecular and cellular puzzle.Am J Otol.1998,19(1):7-19.
18 石磊,张建中,王荣光.胆脂瘤型中耳炎细菌检测.中国耳鼻咽喉.头颈外 科.2004,11(6):366.
19 Vorobev AA, Borisova EV, Borisov VA. Gram-negative virulent bacterial lipopolysaccharide: role in infection and immunity. Vestn Ross Akad Med Nauk.1997, 3:10-13.
20 Peek FA, Huisman MA, Berckmans RJ, etc. Lipopolysaccharide concentration and bone resorption in cholesteatoma. Otol Neurotol. 2003 Sep;24(5):709-13.
21 Dumitrescu AL, Abd-El-Aleem S, Morales-Aza B, etc.A model of periodontitis in the rat: effect of lipopolysaccharide on bone resorption, osteoclast activity, and local peptidergic innervation. J Clin Periodontol. 2004,31(8):596-603.
22 Rogers JE, Li F, Coatney DD,etc.Actinobacillus actinomycetemcomitans lipopolysaccharide-mediated experimental bone loss model for aggressive periodontitis. J Periodontol. 2007 ,78(3):550-558.
23 Deluca HF, Zierold C. Mechnisms and functions of virmain D.Nurt Rev, 1998:56(22):4-10.
24 Carlbeg C, Polly P Gene regulation by vitamin D3. Crit Re Ekuyarot Gene ExPr.l998:8(1):19-42.
25 Holick MF. Vitamin D: A millenium perspective. J Cell Biochem. 2003,88:296-307.
26 Zittermann A. Vitamin D in preventive medicine: are we ignoring the evidence?. Br J Nutr, 2003,89:552-572.
27 Haussler MR, Whitfield GK, Haussler CA, et al. The nuclear vitamin D receptor: biological and molecular regulatory properties revealed. J Bone Miner Res, 1998,13:325-349.
28 Sun J, Kong J, Duan Y, etc. Increased NF-kappaB activity in fibroblasts lacking the vitamin D receptor. Am J Physiol Endocrinol Metab. 2006, 291(2): 315-322.
29 Holick MR Vitamin D: A millenium perspective. J Cell Biochem. 2003,88(2):296-307.
30 Parish JL. Topical vitamin D3 analogues: unapproved uses, dosages, or indications. Clin Dermatol, 2002,20:558-562.
31 Ermis O, Alpsoy E, Cetin L, etc. Is the efficacy of psoralen plus ultraviolet A therapy for vitiligo enhanced by concurrent topical calcipotriol? A placebo-controlled double-blind study. Br J Dermatol, 2001,145:472-5.
32 Inoue M, Matsui T, Nishibu A, etc. Regulatory effectsof l,alpha 25-dihydroxyvitamin D3 on inflammatory responses in psoriasis. Eur J Dermatol, 1998,8:16-20.
33 Pramanik R, Asplin JR, Lindeman C, etc. Lipopolysaccharide negatively modulates vitamin D action by down-regulating expression of vitamin D-induced VDR in human monocytic THP-1 cells. Cell Immunol. 2004,232(1-2): 137-143
34 Timms PM,Mannan N,Hitman GA.Circulating MMP9, vitamin D and variation in the TIMP-1 response with VDR genotype: mechanisms for inflammatory damage in chronic disorders? QJM. 2002,95(12):787-796.
35 Kleiner DE,Stetler-Stevenson WG.Matrix metalloproteinases and metastasis.Cancer Chemother Pharmacol.1999,43:42-51.
36 Wojtowicz-Praga SM,Dickson RB,Hawkins MJ.Matrix metalloproteinase inhibitors.Invest New Drugs.1997,15(1):61-75.
37 Dallari S,Cavani A,Bergamini G etc.Integrin expression in middle ear cholesteatoma.Acta Otolaryngol.1994,114(2):188-192.
38 Moriyama H,Honda Y,Huang CC etc.Bone resorption in cholesteatoma:epithelial-mesenchymal cell interaction and collagenase production.Laryngoscope.1987,97:854-859.
39 Sudhoff H,Bujia J,Borkowski G,etc.Basement membrane in middle ear cholesteatoma immunohistochemical and ultrastructural observations.Ann Otol Rhinol Laryngol.1996,105:804-810.
1 Macri JR,Chole RA.Bone erosion in experimental cholesteatoma--the effects of implanted barriers[J].Otolaryngol Head Neck Surg,1985,93(1):3-17.
2 Chole RA,McGinn MD,Tinling SP.Pressure-induced bone resorption in the middle ear.Ann Otol Rhinol Laryngol.1985 Mar-Apr;94(2 Pt 1):165-70.PMID:3994236.
3 Burger EH,Klein-Nulen J.Responses of bone cells to biomechanical forces in vitro[J].Adv Dent Res,1999,13(1):93-98.
4 Park HJ,Park K.Expression of Fas/APO-1 and apoptosis of keratinocytes in human cholesteatoma[J].Laryngoscope,1999,109(4):613-616.
5 Park K,Chun YM,Lee DH.Expression of phospholipase C-gamma1 in experimental cholesteatoma using Mongolian gerbils[J].Acta Otolaryngol,2001,121(4):477-480.
6 孙文忠,徐志文,唐安洲,等.PTKs、CDK4及p15在中耳胆脂瘤上皮的表达[J].临床耳鼻咽喉科杂志,2004,18(10):616-619.
7 Suzuki C,Ohtani I.Bone destruction resulting from rupture of a cholesteatoma sac:temporal bone pathology[J].Otol Neurotol,2004,25(5):674-677.
8 Takahashi N.Bone and bone related biochemical examinations.Bone and collagen related metabolites.Regulatory mechanisms of osteoclast differentiation and function[J].Clin Calcium,2006,16(6):940-947.
9 Wada T,Nakashima T,Hiroshi N,et al.RANKL-RANK signaling in osteoclastogenesis and bone disease[J].Trends Mol Med,2006,12(1):17-25.
10 Lin JM,Callon KE,Lin CQ,et al.Alteration of bone cell function by RANKL and OPG in different in vitro models[J].Eur J Clin Invest,2007, 37(5): 407-415.
11 Uno Y, Saito R. Bone resorption in human cholesteatoma: morphological study with scanning electron microscopy[J].Ann Otol Rhinol Laryngol, 1995, 104(6) :463-468.
12 Jeong JH, Park CW, Tae K. Expression of RANKL and OPG in middle ear cholesteatoma tissue[J]. Laryngoscope,2006,116(7):1180-1184.
13 Lee ZH, Lee SE, Kim CW,et al. IL-1alpha stimulation of osteoclast survival through the PI 3-kinase/Akt and ERK pathways[J]. J Biochem (Tokyo), 2002, 131(1): 161-166.
14 Bujia J, Kim C, Ostos P, et al. Role of interleukin 6 in epithelial hyperproliferation and bone resorption in middle ear cholesteatomas[J]. Eur Arch Otorhinolaryngol, 1996, 253(3): 152-157.
15 Mihara M, Nishimoto N. Interleukin-6 in pathogenesis of rheumatoid arthritis[J].Nippon Rinsho,2005,63(1):158-162.
16 Kitaura H, Zhou P,Kim HJ,et al.M-CSF mediates TNF-induced inflammatory osteolysis[J].J Clin Invest,2005,115(12):3418-3427.
17 Zhang YH,Heulsmann A,Tondravi MM,et al.Tumor necrosis factor-alpha (TNF) stimulates RANKL-induced osteoclastogenesis via coupling of TNF type 1 receptor and RANK signaling pathways[J].J Biol Chem,2001,276(1):563-568.
18 Hotokezaka H,Sakai E, Ohara N,et al. Molecular analysis of RANKL-independent cell fusion of osteoclast-like cells induced by TNF-alpha,lipopolysaccharide,or peptidoglycan[J]. J Cell Biochem, 2007, 101(1): 122-134.
19 Chole RA.Osteoclasts in chronic otitis media,cholesteatoma,and otosclerosis[J].Ann Otol Rhiool Laryngol,1988,97(6):661-666.
20 Abramson M.Collagenase in the mechanism of action of cholesteatoma[J].Ann Otol Rhinol Laryngol,1971,80(3):414.
21 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.
22 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,2001,8(4):241-244.