【作者】 姚冬梅；

【导师】 濮德敏；

【作者基本信息】 华中科技大学 ， 妇产科学， 2007， 博士

【摘要】 第一部分武汉地区宫颈疾病分布情况的临床调查目的了解武汉地区宫颈疾病的分布情况,为研究宫颈疾病的发病机制、诊治及预防提供临床资料。方法对2004年12月至2005年5月在同济医院妇产科门诊就诊的2635例慢性宫颈炎患者行TCT检查和TBS细胞学分类诊断,对TCT检查发现异常的患者进行阴道镜病理检查和HC-ⅡHPV DNA检测。结果2635例TCT涂片中检出异常涂片493例(18.71%) ,其中不典型鳞状细胞(ASCUS)271例(10.28%) ,低度鳞状上皮内病变(LSIL) 94例(3.57%) ,高度鳞状上皮内病变(HSIL) 14例( 0.53%),鳞状细胞癌(SCC) 1例(0.04% ) ,子宫内膜腺癌1例(0.04%)。阴道镜病检CINⅠ、CINⅡ、CINⅢ检出率分别为31.18% ( 53 /170 )、23.53% ( 40 /170 )、7.06% ( 12 /170 )。TCT示HPV感染共105例,行HPV-DNA检测其中高危型HPV阳性者74例,占70.48%。结论宫颈疾病是育龄妇女的常见病,其中宫颈上皮不典型增生及高危型HPV感染是困扰女性生殖健康的主要元凶。第二部分中药二黄散治疗CINⅠ,Ⅱ合并高危型HPV感染的临床观察及其对血清中IFN-γ,TNF-α,IL-4和IL-10的影响目的:观察中药二黄散治疗CINⅠ,Ⅱ合并高危型HPV感染的临床疗效及其对血清中IFN-γ,TNF-α,IL-4和IL-10的影响。方法:CINⅠ,Ⅱ合并高危型HPV感染158例,其中94例采用二黄散治疗3个月,30例采用α-2b干扰素栓进行对照治疗及34例空白对照,应用二代免疫杂交捕获技术(HC-Ⅱ)于治疗前后分别检查宫颈涂片中高危型HPV-DNA及阴道镜病理检查。抽取二黄散治疗组患者血液,用ELISA方法检测用中药二黄散治疗前后血液中IFN-γ,TNF-α,IL-4和IL-10变化情况。结果:二黄散与α-2b干扰素栓对高危型HPV感染的治疗作用有效率分别为75.6%,73.3%。对于CINⅠ,Ⅱ的治疗有效率为72.4%及73.3%。二者作用相近但明显高于未用药治疗组。CINⅠ,Ⅱ合并高危型HPV感染患者用中药二黄散治疗后,IFN-γ,TNF-α升高而IL-4,IL-10下降。结论:二黄散对于宫颈上皮内瘤变有一定治疗作用。二黄散能使高危型HPV感染者Th1类细胞因子升高,Th2类细胞因子降低,使细胞免疫占优势,从而有利于病毒的清除。第三部分中药二黄散对大鼠子宫颈炎病理形态学的作用及对大鼠宫颈TNF-α,IL-10表达的影响目的:探讨大鼠宫颈炎模型的建立方法,以有效建立宫颈炎模型,为基础及临床研究宫颈炎提供一种有用动物模型。评价中药二黄散对于大鼠宫颈炎病理形态学的作用,观察使用中药治疗前后大鼠宫颈局部TNF-α,IL-10表达的变化情况,初步探讨中药二黄散对大鼠宫颈免疫功能的影响作用。方法:1.建立实验大鼠宫颈炎模型,将20%苯酚浆胶注入实验大鼠阴道内,1次/2日,共5次。2.造模成功后第2天,将造模组大鼠分3组,分别给予相当于成人用量30倍的二黄散,消糜栓(阳性对照)和空白基质(阴性对照),均隔日给药1次,共12次。此外,将未经任何处理的大鼠作为正常对照组。观察大鼠整体状况及阴道分泌物性状。治疗结束后次日处死动物,取子宫观察,宫颈常规石蜡切片行HE染色并在显微镜下观察宫颈形态学变化及炎症程度。3.分别采用免疫组织化学(S-P法),逆转录-聚合酶链反应(RT-PCR)检测中药二黄散用药前后大鼠宫颈TNF-α,IL-10蛋白和mRNA表达。结果:1.病理检查结果:中药二黄散组和消糜栓组(阳性对照组)与阴性对照组相应部位比较,分泌物及炎症程度明显减轻,炎症细胞浸润明显减少,比较有显著性差异(P<0.05),与正常对照组无显著性差异。阴性对照组大鼠阴道外口可见浓稠的分泌物。动物扎堆、少动、竖毛,显微镜下见该组大鼠子宫炎症程度以及粘膜炎细胞数与正常对照组相比较,具有显著性差异。2.免疫组织化学方法检测大鼠宫颈中TNF-α的表达,正常组、阴性对照组、二黄散组吸光度值分别为0.1436±0.0027、0.1769±0.0035、0.1443±0.0012,阴性对照组较正常组明显升高(P<0.01)。二黄散组显著低于阴性对照组(P<0.01)。正常组、阴性对照组、二黄散组中IL-10的吸光度值分别为0.1336±0.0031、0.1386±0.0049、0.1547±0.0051。阴性对照组比正常组表达升高,二黄散组与阴性对照组相比其表达上升,差异具有显著性(P<0.01)。3.RT-PCR技术检测治疗后大鼠宫颈组织中TNF-αmRNA的表达,正常对照组、阴性对照组、二黄散组值分别为0.103±0.031,0.979±0.045,0.719±0.026,阴性对照组较正常对照组明显升高(P<0.01),二黄散组较阴性对照组下降(P<0.01)。IL-10mRNA在正常组、阴性对照组、二黄散组中分别为0.172±0.019、0.487±0.026、0.726±0.021,二黄散组较阴性对照组表达明显升高(P<0.01)。结论:1.采用阴道苯酚胶浆注射可使大鼠宫颈发生炎症改变,其阴道分泌物增多,宫颈炎症程度及炎细胞数均较正常升高,符合宫颈炎症的表现,成功建立宫颈炎模型。2.二黄散能使宫颈病理形态学变化向正常转化。3.大鼠宫颈炎症发生后,宫颈局部TNF-α,IL-10均有升高。中药二黄散治疗可使TNF-α表达下降, IL-10表达升高,从而调节促炎因子和抗炎因子的平衡,有利于宫颈炎症的消散。更多还原

【Abstract】 PartⅠThe clinical investigation on cervical disease of WuhanObjective To investigate the distribution of cervical disease of Wuhan to provide the clinical information for the research of the therapy and prevention of cervical disease.Methods A total of 2634 patients in our hospital had been examined by TCT and TBS from December 2004 toMay 2005.The The abnormal cases which had been selected by TCT were detected by colposcopical pathological examination and Hybrid CaptureⅡHPV testing.Results 493 ( 18.71% ) abnormal smears were found out of 2635 cases, in which atypical squamous cell of undetermined significance (ASCUS) 271cases (10.28% ),low grade squamous intraep ithelial lesion (LSIL) 94 cases ( 3.57% ),high grade squamous intraep ithelial lesion (HSIL) 14 cases (0.53% ) , squamous cell carcinoma 1 case (0.04% ) , adenocarcinoma endometrium 1 case (0104% ).CINⅠ,CINⅡ,CINⅢcases which were diagnosed by colposcopical pathological examination were 31.18% (53 /170 )、23.53% (40 /170 )、7.06% (12 /170 ).HPV infection which was detected by TCT were 105 cases (3.98% ).High risk HPV-positive cases which were detected by Hybrid CaptureⅡHPV testing were 74 cases.Conclusion The disorder of cervical is the normal disease of women.The CIN and HPV infection are the dominant factors which damage the healthy of women. PartⅡThe clinical observation of the effect of Chinese traditional medicine -ErHuangSan on high risk HPV infection CINⅠ,Ⅱand and the expression of IFN-γ,TNF-α, IL-4,IL-10Objective To observe the clinical effect of ErHuangSan on high risk HPV infection CINⅠ,Ⅱand the expession of IFN-γ,TNF-α, IL-4,IL-10.Methods 158 cases of high risk HPV infection patients with CINⅠ,Ⅱwere employed in the study for 3 months. 94 cases were treated by Er Huang San .30 cases were treated byα-2b interferon pessus as the positive control .34 cases were treated by nothing as the blank control.All the cases were detected by Hybrid Cap tureⅡHPV testing, colposcopical pathological examination before and after the treatment.The ELISA was employed to detect the expression of IFN-γ,TNF-αIL-4,IL-10 in Erhuangsan group before and after the administering of ErHuangSan.Results The therapeutic efficacy of ErHuangSan andα-2b interferon pessus on high risk HPV infection were 75.6% and 73.3% respectively. The therapeutic efficacy on CINⅠ,Ⅱwere 72.4% and 73.3%。The The therapeutic efficacy of ErHuangSan andα-2b interferon pessus had no significant difference and better than the blank group obviously.After the treatment of ErHuangSan,the expression of IFN-γand TNF-αraised but the IL-4 and IL-10 deduced.Conclusion ErHuangSan has the therapeutic efftect on the cervical intraepithelial neopasia. ErHuangSan can raise the expression of Th1 cytokine,so that the cellular immunity predominate to eliminate the virus. PartⅢThe effect of Chinese traditional medicine-ErHuangSan on the morphopathology and the expression of TNF-α,IL-10 in cervical tissues of cervicitis ratsObjective To investigate the effective method to build an animal model of cervicitis for the fundamental and clinical research of cervicitis. To investigate the effect of ErHuangSan on the morphopathology and the expression of TNF-α,IL-10 in cervical tissues of cervicitis rats before and after the treatment of ErHuangSan.Preliminary investigate the effect of ErHuangSan on local immunologic function of cervical tissues of Wistar rats.Methods The vagina of experimental Wistar rats were injected the 20% mucilage of phenol to build the cervicitis model,the cervicitis rats were divided randomly to three groups: negative control group, ErHuangSan group and XiaoMiShuan(positive control) group which were treated by blank mucilage ,ErHuangSan and XiaoMiShuan respectively.The rats never be treated were employed as the blank control.the manifestation and the vaginal secretion of rats were observed.The cervical slices were HE stained, the morphological change and the inflammation level were observed by microscope.3.The immunohistochemistry(S-P) and reverse transcription-polymerase chain raction(RT-PCR) were employed to detect the expression of TNF-α,IL-10 in cervical tissues.Results The thickness vaginal secretion could be seen at the collar extension of vagina of the negative control group.The animals of negative control group massed, pilo-erection, Compared with the blank control group,the dgree of cervical inflammation and the quanity inflammatory cells of negative control group had the significant difference(P<0.05). Compared with the negative control group, morphopathology of cervical and the quanity of inflammatory cells of the ErHuangSan group and XiaoMiShuan group had the significant difference (P<0.05). there was no diffrence between the blank control group, ErHuangSan group and XiaoMiShuan group. The immunohistochemistry(s-p) and reverse transcription-polymerase chain raction(RT-PCR) were employed to detect the expression of TNF-α.The absorbency in blank control group,negative group and ErHuangSan group were 0.1436±0.0027、0.1769±0.0035、0.1443±0.0012.Compared with blank control group ,the expression of TNF-αin negative control group increased obviously(P<0.01). Compared with negative control group,the expression of TNF-αin ErHuangSan group reduced(P<0.01).The absorbency value of IL-10 in blank control group,negative control group and ErHuangSan group were 0.1336±0.0031、0.1386±0.0049、0.1547±0.0051.The expression of IL-10 in ErHuangSan group increased.there was significant difference between the ErHuangSan group and negative control group(P<0.01). Reverse transcription-polymerase chain raction(RT-PCR) was employed to detect the expression of TNF-αand IL-10.The expression of TNF-αmRNA in blank control group,negative control group and ErHuangSan medicine group were 0.103±0.031,0.979±0.045,0.719±0.026 respectivily Compared with blank control group ,the expression of TNF-αin negative control group increased obviously(P<0.01). Compared with negative control group,the expression of TNF-αin ErHuangSan group reduced(P<0.01).The expreesion of IL-10 were0.172±0.019、0.487±0.026、0.726±0.021,in blank control group,negative control group and ErHuangSan group. The expression of IL-10 in ErHuangSan group increased.there was significant difference between the ErHuangSan group and negative control group(P<0.01).Conclusion: The injection of 20% phenol mucilage to the vagina of experiment rats can result in the inflammation of cervical . The ErHuangSan was conducive to the healing of the cervical. Along with the occurrence of cevititis ,the expression of cytokine TNF-αand IL-10 raised..The traditional Chinese traditional medicine-ErHuangSan can reduce the expression of TNF-αand raise the expression of IL-10 to adjust the immunity balance of cervical which is good for the convalescence of cervicitis.更多还原