单纯解脲支原体、沙眼衣原体感染及混合感染致小鼠宫颈炎的病理学特点分析和意义
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摘要
目的
探讨解脲支原体(UU)、沙眼衣原体(CT)以及混合感染性小鼠宫颈炎的病理学特点,从而提高对UU、CT的致病性的认识。
方法
从56只小鼠中随机选取48只于第二次皮下注射雌激素当天阴道内接种浓度≥10~6copy/g的UU血清3型,7d后取小鼠阴道内分泌物做UU培养,获得UU阳性小鼠32只,培养阴性者被剔除出试验。将培养阳性小鼠随机分为A、B两组,每组16只,其中A组小鼠于分组后用CT血清D型(浓度为10~5 IFU/ml)进行阴道内再接种,B组小鼠不接种CT。未进行UU接种的剩余8只小鼠设为C组,于第二次皮下注射雌激素后7d,阴道内接种浓度为10~5 IFU/ml的CT血清D型。21d后处死全部小鼠,取宫颈分泌物行UU、CT荧光定量PCR检测并统计感染情况。观察小鼠宫颈病变并做病理学分析,同时用扫描电镜和透射电镜检测。
结果
(1)单纯感染UU或CT与混合感染UU+CT在行为表现无特异性。
(2)本试验首次成功获得同时感染UU+CT且表现为宫颈炎的小鼠模型。
(3)本实验在A组获得9只存在混合感染的小鼠,B组有7只单纯感染UU的小鼠,C组有4只单纯感染CT的小鼠。
(4)单纯UU感染性宫颈炎表现以淋巴细胞浸润为主;单纯CT感染性宫颈炎表现以淋巴细胞、浆细胞浸润为主的滤泡性炎;混合感染性宫颈炎表现具有多形性,病变加重。
(5)扫描电镜下,在单纯感染UU的宫颈管黏膜上皮细胞表面可见球杆状支原体;单纯感染CT的宫颈管黏膜上皮细胞表面未能探及病原体;混合感染的宫颈管黏膜上皮细胞及纤毛微绒毛损伤严重,于细胞表面絮状物覆盖下可探及大量球形衣原体及丝状支原体。
(6)透射电镜下,单纯UU感染时上皮细胞排列整齐,纤毛脱落,具有分叶核的中性粒细胞浸润;单纯CT感染时吞噬细胞内见衣原体原体,少量纤维母细胞增生;混合感染时上皮损伤严重,多形核白细胞浸润,巨噬细胞内见含各期沙眼衣原体的包涵体;而空白对照组超微结构无明显异常。
结论
单纯UU或CT感染与UU+CT混合感染致宫颈炎病理学和超微病理学特点有差异,混合感染病变明显加重。
Objective
To analyse the pathological characteristic of murine cervicitis infected by the only ureaplasma urealyticum(UU) or the only chlamydia trachomatis(CT) and the two mixed microorganism.In the purpose of masterying more knowledges about the pathogenicity of the two kind of microorganism.
Method
From 56 female mices,48 were selected in random to be inoculated with UU serotpe 3 with concentration moer than 10~6copy/g just in the day of treatment with estradiol. After 7 days,secretion from those murine canals were collected for UU culture. And we got 32 UU positive mices.The UU positive mices were divided into two groups ,that's group A and B.There were 16 mices in each group.Mices in group A were further inoculated with CT serotype D from vagino after grouping.In group B, mices were not inoculated with CT serotpe D.And the rest 8 mices that were never inoculated with UU were built up group C.Those mices in group C were only inoculated with CT serotype D after 7 days of treatment with estradiol.21 days later all the mices were killed and cervical swabs were taken so that we had detection by both culture and FQ-PCR and had an analysis on pathogenic microorganism.The tissue of mice cervical canals were retented for pathological research and were detected by both scanning electron microscope(SEM) and transmission electron microscope(TEM).
Result
(1) There is little difference of behavior modification among all the mices.
(2) We successfully find some cervicitis animal models infected by UU+CT.
(3) We find that 9 mices are infected with both UU and CT in group A,and 7 mices are infected only by UU in group B,and 4 mices are infected only by CT in group C.
(4) Non-specific inflammation in sole UU infected cervicitis and cervicitis represented typical follicular cervicitis in sole CT infection and pantomorphic cervicitis in mixed infection.
(5) Under SEM baculiform UU was found on the mucous membrane epithelial cell of cervical canas in the case of sole UU infection, and no microorganism was found but structure badly harmed in the case of sole CT infection, and in the mixed infection there were many spherical CT and filamentous UU hidden in the seriously damaged cilia and microvilli on the superficies of epithelial cell.
(6) Under TEM in sole UU infective cervicitis, epithelial cell alined in order and cilia broke off, and segmented neutrophile granulocytes were found in desmohemoblast;in sole CT infective cervicitis, reticular body(RB) of CT was found in the cytoplasma of macrophagocyte and more segmented neutrophile granulocytes infiltrated desmohemoblast;in the mixed infective cervicitis, epithelia cell was more seriously damaged and the most inflammatory cell whose nucleus exhibited polymorphism and segmented infiltrated desmohemoblast.Each and every periods of CT, including RB、metaphase and maturation phase were found in macrophagocyte which also was full of lysosome.But there wasn't any change of ultrastructure in the control group.
Conclusion
There are differents in the characteristic of murine cervicitis caused by sole UU or CT and the mixed microorganism infected. Moreover the pathological damnification is much severe in the mixed infected mices.
探讨解脲支原体(UU)、沙眼衣原体(CT)以及混合感染性小鼠宫颈炎的病理学特点,从而提高对UU、CT的致病性的认识。
方法
从56只小鼠中随机选取48只于第二次皮下注射雌激素当天阴道内接种浓度≥10~6copy/g的UU血清3型,7d后取小鼠阴道内分泌物做UU培养,获得UU阳性小鼠32只,培养阴性者被剔除出试验。将培养阳性小鼠随机分为A、B两组,每组16只,其中A组小鼠于分组后用CT血清D型(浓度为10~5 IFU/ml)进行阴道内再接种,B组小鼠不接种CT。未进行UU接种的剩余8只小鼠设为C组,于第二次皮下注射雌激素后7d,阴道内接种浓度为10~5 IFU/ml的CT血清D型。21d后处死全部小鼠,取宫颈分泌物行UU、CT荧光定量PCR检测并统计感染情况。观察小鼠宫颈病变并做病理学分析,同时用扫描电镜和透射电镜检测。
结果
(1)单纯感染UU或CT与混合感染UU+CT在行为表现无特异性。
(2)本试验首次成功获得同时感染UU+CT且表现为宫颈炎的小鼠模型。
(3)本实验在A组获得9只存在混合感染的小鼠,B组有7只单纯感染UU的小鼠,C组有4只单纯感染CT的小鼠。
(4)单纯UU感染性宫颈炎表现以淋巴细胞浸润为主;单纯CT感染性宫颈炎表现以淋巴细胞、浆细胞浸润为主的滤泡性炎;混合感染性宫颈炎表现具有多形性,病变加重。
(5)扫描电镜下,在单纯感染UU的宫颈管黏膜上皮细胞表面可见球杆状支原体;单纯感染CT的宫颈管黏膜上皮细胞表面未能探及病原体;混合感染的宫颈管黏膜上皮细胞及纤毛微绒毛损伤严重,于细胞表面絮状物覆盖下可探及大量球形衣原体及丝状支原体。
(6)透射电镜下,单纯UU感染时上皮细胞排列整齐,纤毛脱落,具有分叶核的中性粒细胞浸润;单纯CT感染时吞噬细胞内见衣原体原体,少量纤维母细胞增生;混合感染时上皮损伤严重,多形核白细胞浸润,巨噬细胞内见含各期沙眼衣原体的包涵体;而空白对照组超微结构无明显异常。
结论
单纯UU或CT感染与UU+CT混合感染致宫颈炎病理学和超微病理学特点有差异,混合感染病变明显加重。
Objective
To analyse the pathological characteristic of murine cervicitis infected by the only ureaplasma urealyticum(UU) or the only chlamydia trachomatis(CT) and the two mixed microorganism.In the purpose of masterying more knowledges about the pathogenicity of the two kind of microorganism.
Method
From 56 female mices,48 were selected in random to be inoculated with UU serotpe 3 with concentration moer than 10~6copy/g just in the day of treatment with estradiol. After 7 days,secretion from those murine canals were collected for UU culture. And we got 32 UU positive mices.The UU positive mices were divided into two groups ,that's group A and B.There were 16 mices in each group.Mices in group A were further inoculated with CT serotype D from vagino after grouping.In group B, mices were not inoculated with CT serotpe D.And the rest 8 mices that were never inoculated with UU were built up group C.Those mices in group C were only inoculated with CT serotype D after 7 days of treatment with estradiol.21 days later all the mices were killed and cervical swabs were taken so that we had detection by both culture and FQ-PCR and had an analysis on pathogenic microorganism.The tissue of mice cervical canals were retented for pathological research and were detected by both scanning electron microscope(SEM) and transmission electron microscope(TEM).
Result
(1) There is little difference of behavior modification among all the mices.
(2) We successfully find some cervicitis animal models infected by UU+CT.
(3) We find that 9 mices are infected with both UU and CT in group A,and 7 mices are infected only by UU in group B,and 4 mices are infected only by CT in group C.
(4) Non-specific inflammation in sole UU infected cervicitis and cervicitis represented typical follicular cervicitis in sole CT infection and pantomorphic cervicitis in mixed infection.
(5) Under SEM baculiform UU was found on the mucous membrane epithelial cell of cervical canas in the case of sole UU infection, and no microorganism was found but structure badly harmed in the case of sole CT infection, and in the mixed infection there were many spherical CT and filamentous UU hidden in the seriously damaged cilia and microvilli on the superficies of epithelial cell.
(6) Under TEM in sole UU infective cervicitis, epithelial cell alined in order and cilia broke off, and segmented neutrophile granulocytes were found in desmohemoblast;in sole CT infective cervicitis, reticular body(RB) of CT was found in the cytoplasma of macrophagocyte and more segmented neutrophile granulocytes infiltrated desmohemoblast;in the mixed infective cervicitis, epithelia cell was more seriously damaged and the most inflammatory cell whose nucleus exhibited polymorphism and segmented infiltrated desmohemoblast.Each and every periods of CT, including RB、metaphase and maturation phase were found in macrophagocyte which also was full of lysosome.But there wasn't any change of ultrastructure in the control group.
Conclusion
There are differents in the characteristic of murine cervicitis caused by sole UU or CT and the mixed microorganism infected. Moreover the pathological damnification is much severe in the mixed infected mices.
引文
[1]王淑玉.实用妇产科诊疗规范[M].江苏:江苏科学技术出版社,2002:28-33.
[2]陆良玉.沙眼衣原体解脲支原体感染与输卵管性不孕的关系.浙江临床医学,2004,6(7):594.
[3]Imudia AN,Detti L,Puscheck EE,Yelian FD,Diamond MP.The preva-lence of ureaplasma urealyticum,mycoplasma hominis,chlamydia trachomatis and neisseria gonorrhoeae infections,and the rubella status of patients undergoing an initial infertility evaluation.J Assist Reprod Genet.2008 Jan;25(1):43-6.Epub 2008 Jan 18.
[4]Wang Y,Sun G,Pan JG,Li T.Correlation of Ureaplasma urealytieum and Chlamydia trachomatis infections with male sterility:a meta-analysis of ran-domized control trials.Zhonghua Nan Ke Xue.2006 Jul;12(7):615-8.Chinese.
[5]余慧敏,凌晟荣.粘液脓性宫颈炎与解脲支原体、人型支原体、沙眼衣原体感染关系的临床研究.中国妇幼保健,2007,22:3516-3517.
[6]申建威,孙秀琴,程冬娥.女性生殖道性传播疾病病原体混合感染调查.中华医院感染学杂志,2007,17(5):533-534.
[7]Liu XY,Sun HM,Feng YL,Hu J,Zhao HQ,Zhang LY.Study on the pathogens correlated to sexually transmitted diseases in 285 pre-pubertal girls with vulvovaginitis in Beijing.Zhonghua Liu Xing Bing Xue Za Zhi.2007 Aug;28(8):765-7.
[8]张洪文,陈琳,王新等.解脲支原体生物一群在不同女性人群生殖道中的分布.中国医师杂志,2007,19(2):170-172.
[9]张帝开,覃春容.女性下生殖道解脲支原体的定量检测与致病性研究..中国实用妇科与产科杂志,2005,21(11):656-658.
[10]李范青,蒋义玲,马淑华.宫颈糜烂局部衣原体及支原体的检测及意义.中国实用妇科与产科杂志,2004,20(7):417-418.
[11]张宁,唐轶,李力,等.荧光定量PCR检测宫颈分泌物中解脲支原体.临床检验杂志,2001,19(3):179.
[12]Taylor-Robinson D,Furr PM.Observations on experimental colonisat-ion of mice by ureaplasmas of human origin.J Med Microbiol,2002,51:866-870.
[13]Pal S,Peterson EM,Luis M.et al.Susceptibility of mice to vaginal infection with Chlamydia trachomatis mouse pneumonitis is dependent on the age of the animal [J].Infect Immun,2001,69(8):2067-73.
[14]啜丽清,刘俊芬,张林爱.孕妇沙眼衣原体和解脲支原体感染与稽留流产的关.中国妇幼保健,2005,20:1721-1722.
[15]于红,王蓓,金辉等.解脲脲原体与女性生殖道其他病原体感染的交互作用分析.实用妇产科杂志,2006,22(7):433-435.
[16]武忠弼主编.超微病理诊断学[M].上海:科学技术出版社.2003:730-737
[17]Yu JT,Tang WY,Lau KH,Chong LY,Lo KK.Asymptomatic urethral infection in male sexually transmitted disease clinic attendees.Int J STD AIDS.2008Mar;19(3):155-8.
[18]Li YH,Bratmer A,Jonsson B,et al.Ureaplasma urealytieum induced production of proinflammatory eytokines by macrophages[J].Pediatr Res,2000,48(1):114-119.
[19]Into T,Shibata K.Apoptosis signal regulating kinase 12 mediated sust-ained p38mitogen activated protein kinase activation regulates mycoplasmal lipoprotein and staphylococcal peptidoglycan triggered Toll-like receptor signalling pathways[J].CellMicrobiol,2005,7(9):130521317.
[20]Morrison SG,Morrison RP.In situ analysis of the evolution of the primary immune response in routine chlamydia trachomatis genital tract infection.Infect Immun,2000,68:2870.
[21]Dong F,PirbhaiM,Xiao Y,et al.Degradation of the proapoptoticp roteins Bik,Puma,and Bim with Bcl-2 domain homology in Chlamydia trachomatis infected cells[J].Infect Imm un,2005,73(3):1861-1864.
[22]印洁,施毅,周晓军,等.肺炎衣原体肺部感染的超微病理研究.医学研究生报,2002,15(6):496-500.
[23]Kutlin A,Flegg C,Stenzel D,et al.Ultrastructural study of Chlamydia pneumoniae in a continuous infection model[J].J Clin Mierobiol,2001,39:3721-3723.
[24]HuntM E,Brown D R.Mycoplasma alligatoris infection promotes CD-95(FasR)expression and apoptosis of primary cardiac fibroblasts[J].Clin Diagn Lab Immunol,2005,12(12):1370-1377.
[25]Fdedek D,Ekiel A,Chclmicki Z,et al.HPV,Chlamydia trachomatis and genital mycoplasmas infections in women with low-grade squamous intracpithe-lial lesions(LSIL)[J].Ginekol Pol,2004,75(6):457-463.
[26]Golijow CD,Abba MC,Mouron SA,et al.Chlamydia trachomatis and Human papilloma virus infections in cervical disease in Argentine women[J].Gynecol Oncol,2005,96(1):181-186.
[2]陆良玉.沙眼衣原体解脲支原体感染与输卵管性不孕的关系.浙江临床医学,2004,6(7):594.
[3]Imudia AN,Detti L,Puscheck EE,Yelian FD,Diamond MP.The preva-lence of ureaplasma urealyticum,mycoplasma hominis,chlamydia trachomatis and neisseria gonorrhoeae infections,and the rubella status of patients undergoing an initial infertility evaluation.J Assist Reprod Genet.2008 Jan;25(1):43-6.Epub 2008 Jan 18.
[4]Wang Y,Sun G,Pan JG,Li T.Correlation of Ureaplasma urealytieum and Chlamydia trachomatis infections with male sterility:a meta-analysis of ran-domized control trials.Zhonghua Nan Ke Xue.2006 Jul;12(7):615-8.Chinese.
[5]余慧敏,凌晟荣.粘液脓性宫颈炎与解脲支原体、人型支原体、沙眼衣原体感染关系的临床研究.中国妇幼保健,2007,22:3516-3517.
[6]申建威,孙秀琴,程冬娥.女性生殖道性传播疾病病原体混合感染调查.中华医院感染学杂志,2007,17(5):533-534.
[7]Liu XY,Sun HM,Feng YL,Hu J,Zhao HQ,Zhang LY.Study on the pathogens correlated to sexually transmitted diseases in 285 pre-pubertal girls with vulvovaginitis in Beijing.Zhonghua Liu Xing Bing Xue Za Zhi.2007 Aug;28(8):765-7.
[8]张洪文,陈琳,王新等.解脲支原体生物一群在不同女性人群生殖道中的分布.中国医师杂志,2007,19(2):170-172.
[9]张帝开,覃春容.女性下生殖道解脲支原体的定量检测与致病性研究..中国实用妇科与产科杂志,2005,21(11):656-658.
[10]李范青,蒋义玲,马淑华.宫颈糜烂局部衣原体及支原体的检测及意义.中国实用妇科与产科杂志,2004,20(7):417-418.
[11]张宁,唐轶,李力,等.荧光定量PCR检测宫颈分泌物中解脲支原体.临床检验杂志,2001,19(3):179.
[12]Taylor-Robinson D,Furr PM.Observations on experimental colonisat-ion of mice by ureaplasmas of human origin.J Med Microbiol,2002,51:866-870.
[13]Pal S,Peterson EM,Luis M.et al.Susceptibility of mice to vaginal infection with Chlamydia trachomatis mouse pneumonitis is dependent on the age of the animal [J].Infect Immun,2001,69(8):2067-73.
[14]啜丽清,刘俊芬,张林爱.孕妇沙眼衣原体和解脲支原体感染与稽留流产的关.中国妇幼保健,2005,20:1721-1722.
[15]于红,王蓓,金辉等.解脲脲原体与女性生殖道其他病原体感染的交互作用分析.实用妇产科杂志,2006,22(7):433-435.
[16]武忠弼主编.超微病理诊断学[M].上海:科学技术出版社.2003:730-737
[17]Yu JT,Tang WY,Lau KH,Chong LY,Lo KK.Asymptomatic urethral infection in male sexually transmitted disease clinic attendees.Int J STD AIDS.2008Mar;19(3):155-8.
[18]Li YH,Bratmer A,Jonsson B,et al.Ureaplasma urealytieum induced production of proinflammatory eytokines by macrophages[J].Pediatr Res,2000,48(1):114-119.
[19]Into T,Shibata K.Apoptosis signal regulating kinase 12 mediated sust-ained p38mitogen activated protein kinase activation regulates mycoplasmal lipoprotein and staphylococcal peptidoglycan triggered Toll-like receptor signalling pathways[J].CellMicrobiol,2005,7(9):130521317.
[20]Morrison SG,Morrison RP.In situ analysis of the evolution of the primary immune response in routine chlamydia trachomatis genital tract infection.Infect Immun,2000,68:2870.
[21]Dong F,PirbhaiM,Xiao Y,et al.Degradation of the proapoptoticp roteins Bik,Puma,and Bim with Bcl-2 domain homology in Chlamydia trachomatis infected cells[J].Infect Imm un,2005,73(3):1861-1864.
[22]印洁,施毅,周晓军,等.肺炎衣原体肺部感染的超微病理研究.医学研究生报,2002,15(6):496-500.
[23]Kutlin A,Flegg C,Stenzel D,et al.Ultrastructural study of Chlamydia pneumoniae in a continuous infection model[J].J Clin Mierobiol,2001,39:3721-3723.
[24]HuntM E,Brown D R.Mycoplasma alligatoris infection promotes CD-95(FasR)expression and apoptosis of primary cardiac fibroblasts[J].Clin Diagn Lab Immunol,2005,12(12):1370-1377.
[25]Fdedek D,Ekiel A,Chclmicki Z,et al.HPV,Chlamydia trachomatis and genital mycoplasmas infections in women with low-grade squamous intracpithe-lial lesions(LSIL)[J].Ginekol Pol,2004,75(6):457-463.
[26]Golijow CD,Abba MC,Mouron SA,et al.Chlamydia trachomatis and Human papilloma virus infections in cervical disease in Argentine women[J].Gynecol Oncol,2005,96(1):181-186.