乳牙尖周病损中HCMV和EBV片段表达的研究
摘要
有研究证明,发生在成人症状明显的、大范围的牙根尖周病变与巨细胞病毒(HCMV)及Epstein-Barr(EBV)病毒感染有关。Slots等认为,HCMV和EBV感染可通过诱导局部组织细胞产生致炎性细胞因子、损害局部防御机制、提升局部常驻细菌毒力来造成根尖周病变。然而,有关上述两种病毒与乳牙根尖周病变的关系的研究报道极少。
乳牙根尖周病是乳牙根尖周或根分歧部位的牙骨质,牙周膜和牙槽骨等等组织的炎症性疾病。乳牙根尖周病由于局部组织疏松,血运丰富,极易引起相应颌面部蜂窝织炎,此时有针对性的全身用药就尤为重要,这也需要全面了解根尖周病具体的发病机理。一直以来,人们认为牙髓病和根尖周病主要是由细菌感染引起的,但近几年的研究已经认识到人巨细胞病毒(HCMV),EB病毒(EBV)和其他的一些疱疹病毒与大面积的尖周病损有关。疱疹病毒可能通过直接感染和复制或者引发宿主的防御反应而致病。疱疹病毒介导的牙髓病有可能通过多种机制发生独立或者联合作用,并且可能包括宿主的细胞和体液免疫反应。在成人根尖周病损区均已发现疱疹病毒的存在,研究亦表明HCMV, EBV等疱疹病毒可能与进展性牙周炎以及牙龈和粘膜的急性炎症的发病机理有关。与严重的牙周病相似,学者们预测一些进展性的尖周病损是病毒、细菌与宿主免疫反应相互作用的结果。首先,口腔细菌感染引发的牙髓感染导致病毒感染的炎症细胞进入根管和尖周区域,继发的病毒活化可能加重炎症反应并进一步降低尖周组织的抵抗力,导致病原菌的过度增殖及巨噬细胞和其它宿主细胞分泌一些细胞因子和化学因子,刺激骨吸收。研究亦证实牙源性囊肿中也存在着HCMV。但也有研究认为在根尖周病中虽然有疱疹病毒的存在,但其在根尖周病的发生发展过程中却不是必需的。本研究拟观察HCMV, EBV在乳牙根尖周病变组织和正常牙髓组织中的表达差异,探讨疱疹病毒在乳牙根尖周病发病机理中的作用。
目的：
1.通过PCR技术检测乳牙慢性根尖周病中HCMV, EBV的基因片段。
2.探讨病毒在乳牙根尖周感染进展中的作用及可能的抗病毒治疗的途径。
方法：
1.分别收集有严重根尖病变乳牙的根尖周肉芽组织以及无病变的乳牙正常的牙髓组织。
2.采用PCR技术分别检测两组中HCMV, EBV mRNA的表达。
结果：
1.在有严重根尖周病变乳牙的慢性根尖周肉芽组织中,人巨细胞病毒的检出率是64.3%,EB病毒的检出率是57.1%。
2.在无病变的乳牙正常牙髓组织中,人巨细胞病毒的检出率为7.1%,EB病毒的检出率是14.3%。
3.人巨细胞病毒在两组中的检出率比较差别具有显著性意义(p=0.004<0.05)；EB病毒在两组中的检出率比较差别同样具有显著性意义(p=0.046<0.05)。
结论：
1. HCMV和EBV感染与乳牙根尖周大面积病损有关。
2. HCMV和EBV感染可通过引起直接的细胞毒性反应,损害局部防御机制,刺激炎症细胞分泌促炎细胞因子和化学因子,引起局部组织的免疫反应来造成乳牙根尖周病。
3.由牙髓感染导致的乳牙根尖周病是以细菌感染为主,兼有病毒感染及宿主局部免疫反应的一种多因素感染。
Previous researches showed that a positive relationship exists between periapical human cytomegalovirus and Esptein-Barr virus infections and symptomatic and large-size periapical lesions in adult patients. Slots et al. hypothesized that human cytomegalovirus and Epstein-Barr virus infections may cause periapical pathosis by inducing cytokine and chemokine release from inflammatory or connective tissue cells, or by impairing local host defenses, resulting in heightened virulence of resident bacterial pathogens. However, there is few investigation on the relationship between periapical periodontitis in deciduous tooth and infections by human cytomegalovirus and Esptein-Barr virus.
The periapical pathosis of deciduous teeth is inflammation disease of the cementum, periodontal ligament and alveolar bone in the periapical or root furcation region. Since periapical pathosis of deciduous teeth is extremely easy to cause corresponding maxillofacial cellulitis for its loose tissue and rich blood circulation, it is especially important to give proper medication, which also needs to understand the concrete pathogenesis of periapical pathosis comprehensively. In the past, people thought that endodontic disease and periapical pathosis are mainly caused by the bacterial infection, but the research in recent years has already realized that HCMV, EBV and other herpesvirus may partly be responsible for large periapical lesions. Herpesvirus may cause disease as a direct result of viral infection and replication, or as a result of virally induced impairment of the host defense. Herpesvirus-mediated endodontic disease may take place via several mechanisms, operating alone or in combination, and may involve both cellular and humoral host responses. Herpesvirus family members are all found in periapical lesions in adults. The research also indicates HCMV, EBV and other herpesvirus have been implicated in the pathogenesis of aggressive periodontitis and in acute inflammation of gingiva and oral mucosa. Similarly to severe periodontitis, some scholars hypothesize that some aggressive types of periapical pathosis develop as a result of a series of interactions between bacteria, herpesvirues and host immune reactions. Initially, pulpal infection by oral bacteria causes herpesvirus-infected inflammatory celles to enter the root canal and the periapical region.Subsequent herpesvirus reactivation may then aggravate the inflammatory response and further diminish the resistance of periapical tissue, leading to overgrowth of pathogenic bacteria and release of pro-inflammatory cytokines and chemokines from macrophages and orther host cells to stimulate the bone resorption. And other research also confirmed the existence of HCMV in odontogenic cysts. But many researches also think that herpesvirus are present, but not required for development of periapical lesions. This study intends to investigate the expressive difference of HCMV, EBV in periapical lesion and normal dental pulp of deciduous teeth, to discuss herpesvirus'role in the pathogenesis of periapical lesion of deciduous teeth.
Objective:
1. To examine the gene fragment of HCMV, EBV in chronic periapical lesion through PCR assay.
2. To discuss the role plaid by human cytomegalovirus and Esptein-Barr virus in the progresses of periapical infection of deciduous teeth and the possible antiviral treatment for the disease.
Methods:
1. To collect tissue in seriously infective periapical granulation of deciduous teeth and normal dental pulp respectively.
2. To detect the expression of HCMV, EBV mRNA by PCR method.
Results:
1. In serious chronic periapical granulation of deciduous teeth,the detection rate of HCMV is 64.3%, and the detection rate of EBV is 57.1%.
2. In normal dental pulp of deciduous teeth, the detection rate of HCMV is 7.1%, and the detection rate of EBV is 14.3%.
3. In the two groups, the distinction of the detection rates of HCMV is significant (p=0.004<0.05); And that of EBV is also significant (p=0.046<0.05)
Conclusion:
1. A positive relationship exists between HCMV and EBV infections and large-size periapical lesions in deciduoud teeth.
2. HCMV and EBV infections may cause periapical pathosis in deciduous teeth by causing direct cytopathic effects and by impairing local host defenses, or by eliciting proflammatory cytokine and chemokine release from inflammatory cells and by inducing host immune responses.
3. The periapical pathosis of deciduous teeth caused by pulp infection is the multiple factor infection mainly because of bacterial infection and partly viral infection and host immune responses.
乳牙根尖周病是乳牙根尖周或根分歧部位的牙骨质,牙周膜和牙槽骨等等组织的炎症性疾病。乳牙根尖周病由于局部组织疏松,血运丰富,极易引起相应颌面部蜂窝织炎,此时有针对性的全身用药就尤为重要,这也需要全面了解根尖周病具体的发病机理。一直以来,人们认为牙髓病和根尖周病主要是由细菌感染引起的,但近几年的研究已经认识到人巨细胞病毒(HCMV),EB病毒(EBV)和其他的一些疱疹病毒与大面积的尖周病损有关。疱疹病毒可能通过直接感染和复制或者引发宿主的防御反应而致病。疱疹病毒介导的牙髓病有可能通过多种机制发生独立或者联合作用,并且可能包括宿主的细胞和体液免疫反应。在成人根尖周病损区均已发现疱疹病毒的存在,研究亦表明HCMV, EBV等疱疹病毒可能与进展性牙周炎以及牙龈和粘膜的急性炎症的发病机理有关。与严重的牙周病相似,学者们预测一些进展性的尖周病损是病毒、细菌与宿主免疫反应相互作用的结果。首先,口腔细菌感染引发的牙髓感染导致病毒感染的炎症细胞进入根管和尖周区域,继发的病毒活化可能加重炎症反应并进一步降低尖周组织的抵抗力,导致病原菌的过度增殖及巨噬细胞和其它宿主细胞分泌一些细胞因子和化学因子,刺激骨吸收。研究亦证实牙源性囊肿中也存在着HCMV。但也有研究认为在根尖周病中虽然有疱疹病毒的存在,但其在根尖周病的发生发展过程中却不是必需的。本研究拟观察HCMV, EBV在乳牙根尖周病变组织和正常牙髓组织中的表达差异,探讨疱疹病毒在乳牙根尖周病发病机理中的作用。
目的：
1.通过PCR技术检测乳牙慢性根尖周病中HCMV, EBV的基因片段。
2.探讨病毒在乳牙根尖周感染进展中的作用及可能的抗病毒治疗的途径。
方法：
1.分别收集有严重根尖病变乳牙的根尖周肉芽组织以及无病变的乳牙正常的牙髓组织。
2.采用PCR技术分别检测两组中HCMV, EBV mRNA的表达。
结果：
1.在有严重根尖周病变乳牙的慢性根尖周肉芽组织中,人巨细胞病毒的检出率是64.3%,EB病毒的检出率是57.1%。
2.在无病变的乳牙正常牙髓组织中,人巨细胞病毒的检出率为7.1%,EB病毒的检出率是14.3%。
3.人巨细胞病毒在两组中的检出率比较差别具有显著性意义(p=0.004<0.05)；EB病毒在两组中的检出率比较差别同样具有显著性意义(p=0.046<0.05)。
结论：
1. HCMV和EBV感染与乳牙根尖周大面积病损有关。
2. HCMV和EBV感染可通过引起直接的细胞毒性反应,损害局部防御机制,刺激炎症细胞分泌促炎细胞因子和化学因子,引起局部组织的免疫反应来造成乳牙根尖周病。
3.由牙髓感染导致的乳牙根尖周病是以细菌感染为主,兼有病毒感染及宿主局部免疫反应的一种多因素感染。
Previous researches showed that a positive relationship exists between periapical human cytomegalovirus and Esptein-Barr virus infections and symptomatic and large-size periapical lesions in adult patients. Slots et al. hypothesized that human cytomegalovirus and Epstein-Barr virus infections may cause periapical pathosis by inducing cytokine and chemokine release from inflammatory or connective tissue cells, or by impairing local host defenses, resulting in heightened virulence of resident bacterial pathogens. However, there is few investigation on the relationship between periapical periodontitis in deciduous tooth and infections by human cytomegalovirus and Esptein-Barr virus.
The periapical pathosis of deciduous teeth is inflammation disease of the cementum, periodontal ligament and alveolar bone in the periapical or root furcation region. Since periapical pathosis of deciduous teeth is extremely easy to cause corresponding maxillofacial cellulitis for its loose tissue and rich blood circulation, it is especially important to give proper medication, which also needs to understand the concrete pathogenesis of periapical pathosis comprehensively. In the past, people thought that endodontic disease and periapical pathosis are mainly caused by the bacterial infection, but the research in recent years has already realized that HCMV, EBV and other herpesvirus may partly be responsible for large periapical lesions. Herpesvirus may cause disease as a direct result of viral infection and replication, or as a result of virally induced impairment of the host defense. Herpesvirus-mediated endodontic disease may take place via several mechanisms, operating alone or in combination, and may involve both cellular and humoral host responses. Herpesvirus family members are all found in periapical lesions in adults. The research also indicates HCMV, EBV and other herpesvirus have been implicated in the pathogenesis of aggressive periodontitis and in acute inflammation of gingiva and oral mucosa. Similarly to severe periodontitis, some scholars hypothesize that some aggressive types of periapical pathosis develop as a result of a series of interactions between bacteria, herpesvirues and host immune reactions. Initially, pulpal infection by oral bacteria causes herpesvirus-infected inflammatory celles to enter the root canal and the periapical region.Subsequent herpesvirus reactivation may then aggravate the inflammatory response and further diminish the resistance of periapical tissue, leading to overgrowth of pathogenic bacteria and release of pro-inflammatory cytokines and chemokines from macrophages and orther host cells to stimulate the bone resorption. And other research also confirmed the existence of HCMV in odontogenic cysts. But many researches also think that herpesvirus are present, but not required for development of periapical lesions. This study intends to investigate the expressive difference of HCMV, EBV in periapical lesion and normal dental pulp of deciduous teeth, to discuss herpesvirus'role in the pathogenesis of periapical lesion of deciduous teeth.
Objective:
1. To examine the gene fragment of HCMV, EBV in chronic periapical lesion through PCR assay.
2. To discuss the role plaid by human cytomegalovirus and Esptein-Barr virus in the progresses of periapical infection of deciduous teeth and the possible antiviral treatment for the disease.
Methods:
1. To collect tissue in seriously infective periapical granulation of deciduous teeth and normal dental pulp respectively.
2. To detect the expression of HCMV, EBV mRNA by PCR method.
Results:
1. In serious chronic periapical granulation of deciduous teeth,the detection rate of HCMV is 64.3%, and the detection rate of EBV is 57.1%.
2. In normal dental pulp of deciduous teeth, the detection rate of HCMV is 7.1%, and the detection rate of EBV is 14.3%.
3. In the two groups, the distinction of the detection rates of HCMV is significant (p=0.004<0.05); And that of EBV is also significant (p=0.046<0.05)
Conclusion:
1. A positive relationship exists between HCMV and EBV infections and large-size periapical lesions in deciduoud teeth.
2. HCMV and EBV infections may cause periapical pathosis in deciduous teeth by causing direct cytopathic effects and by impairing local host defenses, or by eliciting proflammatory cytokine and chemokine release from inflammatory cells and by inducing host immune responses.
3. The periapical pathosis of deciduous teeth caused by pulp infection is the multiple factor infection mainly because of bacterial infection and partly viral infection and host immune responses.
引文
1.石四箴主编.儿童口腔医学.第一版.北京：人民卫生出版社,2001,108-109.
2. Sabeti M, Vslles Y, Nowzari H, et al. Cytomegalovirus and Epstein-Barr virus DNA transcription in endodontic symptomatic lesions. Oral Microbiology Immunology,2003,18(6):104-108.
3. Slots J, Sabeti M, Simon JH. Herpesviruses in periapical pathosis:an etiopathogenic relationship?Oral Surgery Oral Medicine Oral Pathology Oral Radiology &Endodontics.2003,96(3):327-331.
4. Yildirim S, Yapar M, Kubar A, et al. Human cytomegalovirus, Epstein-Barr virus and bone resorption-inducing cytokines in periapical lesions of deciduous teeth. Oral Microbiol Immunol. 2006,21 (2):107-111.
5. Contreras A,Slots J.Herpesviruse in human periodontal disease.J Periodontal Res,2000,35(8):3-16.
6. Slots J.Interactions between herpesvirus and bacteria in human periodontal diseases In:Brogden KA,Guthmiller JM, eds. Polymicrobial diseasas,2002,16(5):317-331.
7. Andrcic M, Milasin J, Jovanovic T, rt al.Human Cytomegalovirus is present in odontogenic cysts. Oral Mirobiology Immunology, 2007,22(9):347-351.
8. Vicky Chen, Yanwen Chen, Hong Li, et al. Herpesviruses in Abscesses and Cellulitis of Endodontic Origin. J Endod.2009,35(2):182-188.
9.樊明文主编.牙体牙髓病学.第二版.北京：人民卫生出版社,2003,150-151.
10.杨秋波,樊鲁娜,时清.乳牙急性根尖周脓肿细菌的DGGE分析.北京口腔医学.2009,17(4)：223-225.
11.于金华,俞未一,刘卫红.替硝唑治疗诱发性大鼠根尖周炎的组织学分析.临床口腔医学杂志,2003,19(10)：24-25.
12.王艳华,侯本祥.感染根管非细菌微生物学的研究进展.现代口腔医学杂志.2009,23(1)：86-87.
13. Slots J, Nowzari H, Sabeti M. Cytomegalovirus infection in symptomatic periapical pathosis. Int Endod J.2004,37(8):519-524.
14. Saygun I, Yapar M, Ozdemir A, Kubar A, Slots J. Human Cytomegalovirus and Epstein-Barr virus type 1 in periodontal abscesses. Oral Microbiol Immunol.2004,19(2):83-87.
15. Sunde PT, Olsen I, Enersen M, Beiske K, Grinde B. Human cytomegalovirus and Epstein-Barr virus in apical and marginal periodontitis:a role in pathology?J Med Virol.2008,80(6): 1007-1011.
16.关为群,林霏,周文娟等.龈下刮治前后不同牙周状态下龈下菌斑中HCMV病毒的检测.福建医科大学学报,2007,41(5)：454-456.
17.吉林大学基础医学院病原生物学教研室.人类疱疹病毒教学课件：2-31.
18.王延军,孙斌,刘道洁等.巨细胞病毒感染细胞的免疫原性分析.细胞与分子免疫学杂志,2009,25(10)907-909.
19.曾宾,肖燕,金润铭等.儿童EB病毒感染的免疫功能状况.实用儿科临床杂志,2006,21(22)：1549-1550.
20.尚姝环,李成章,樊明文.疱疹病毒感染与牙周炎.国外医学：口腔医学分册,2004,31(2)：115-116.
21.郝鹏杰,张晓明.破骨细胞在口腔正畸领域中的研究现状.滨州医学院学报,2007,30(2)：121-123.
22.曹采方主编.牙周病学.第二版,北京：人民卫生出版社,2003,64-66.
23. Saboia-Dantas CJ, Coutrin de Toledo LF, Sampaio-Filho HR, et al. Herpesviruses in asymptomatic apical periodontitis lesions:an immunohistochemical approach. Oral Microbiol Immunol.2007,22(5): 320-325.
24. Botero JE, Vidal C, Contreras A, et al. Comparison of nested polymerase chain reaction (PCR), real-time PCR and viral culture for the detection of cytomegalovirus in subgingival samples. Oral Microbiol Immunol.2008,23(3):239-244.
25. Grinde B. Human circoviruses:Clincal and applied perspectives. Curr Top Virol.2003,3:81-90.
2. Sabeti M, Vslles Y, Nowzari H, et al. Cytomegalovirus and Epstein-Barr virus DNA transcription in endodontic symptomatic lesions. Oral Microbiology Immunology,2003,18(6):104-108.
3. Slots J, Sabeti M, Simon JH. Herpesviruses in periapical pathosis:an etiopathogenic relationship?Oral Surgery Oral Medicine Oral Pathology Oral Radiology &Endodontics.2003,96(3):327-331.
4. Yildirim S, Yapar M, Kubar A, et al. Human cytomegalovirus, Epstein-Barr virus and bone resorption-inducing cytokines in periapical lesions of deciduous teeth. Oral Microbiol Immunol. 2006,21 (2):107-111.
5. Contreras A,Slots J.Herpesviruse in human periodontal disease.J Periodontal Res,2000,35(8):3-16.
6. Slots J.Interactions between herpesvirus and bacteria in human periodontal diseases In:Brogden KA,Guthmiller JM, eds. Polymicrobial diseasas,2002,16(5):317-331.
7. Andrcic M, Milasin J, Jovanovic T, rt al.Human Cytomegalovirus is present in odontogenic cysts. Oral Mirobiology Immunology, 2007,22(9):347-351.
8. Vicky Chen, Yanwen Chen, Hong Li, et al. Herpesviruses in Abscesses and Cellulitis of Endodontic Origin. J Endod.2009,35(2):182-188.
9.樊明文主编.牙体牙髓病学.第二版.北京：人民卫生出版社,2003,150-151.
10.杨秋波,樊鲁娜,时清.乳牙急性根尖周脓肿细菌的DGGE分析.北京口腔医学.2009,17(4)：223-225.
11.于金华,俞未一,刘卫红.替硝唑治疗诱发性大鼠根尖周炎的组织学分析.临床口腔医学杂志,2003,19(10)：24-25.
12.王艳华,侯本祥.感染根管非细菌微生物学的研究进展.现代口腔医学杂志.2009,23(1)：86-87.
13. Slots J, Nowzari H, Sabeti M. Cytomegalovirus infection in symptomatic periapical pathosis. Int Endod J.2004,37(8):519-524.
14. Saygun I, Yapar M, Ozdemir A, Kubar A, Slots J. Human Cytomegalovirus and Epstein-Barr virus type 1 in periodontal abscesses. Oral Microbiol Immunol.2004,19(2):83-87.
15. Sunde PT, Olsen I, Enersen M, Beiske K, Grinde B. Human cytomegalovirus and Epstein-Barr virus in apical and marginal periodontitis:a role in pathology?J Med Virol.2008,80(6): 1007-1011.
16.关为群,林霏,周文娟等.龈下刮治前后不同牙周状态下龈下菌斑中HCMV病毒的检测.福建医科大学学报,2007,41(5)：454-456.
17.吉林大学基础医学院病原生物学教研室.人类疱疹病毒教学课件：2-31.
18.王延军,孙斌,刘道洁等.巨细胞病毒感染细胞的免疫原性分析.细胞与分子免疫学杂志,2009,25(10)907-909.
19.曾宾,肖燕,金润铭等.儿童EB病毒感染的免疫功能状况.实用儿科临床杂志,2006,21(22)：1549-1550.
20.尚姝环,李成章,樊明文.疱疹病毒感染与牙周炎.国外医学：口腔医学分册,2004,31(2)：115-116.
21.郝鹏杰,张晓明.破骨细胞在口腔正畸领域中的研究现状.滨州医学院学报,2007,30(2)：121-123.
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