摘要目的 研究沙眼衣原体(Chlamydia trachomatis,Ct)pORF5质粒蛋白对小鼠生殖道组织的免疫损伤作用,并初步探讨其损伤机制.方法 表达纯化GST-pORF5融合蛋白,融合蛋白经PreScission Protease酶切和去内毒素处理后分别在第1、3、6天接种于BALB/c小鼠阴道后穹窿,第7天处死小鼠,分离小鼠生殖道组织,肉眼大体观察并进行炎症病理评分；ELISA方法检测血清、阴道灌洗液和小鼠脾细胞培养上清中的TNF-α、IL-1β和IL-6细胞因子水平.结果 pORF5蛋白接种组小鼠输卵管壶腹部与峡部出现不同程度的肿胀,周围结缔组织发生黏连,并出现了不同程度的扭曲与积水,而PBS和GST(谷胱甘肽S-转移酶)蛋白对照组输卵管组织未出现明显改变；同时生殖道组织炎症病理积分明显高于PBS对照组(P＜0.01)和GST对照组(P＜0.01)；pORF5蛋白接种组小鼠脾细胞培养上清、阴道分泌物中TNF-α、IL-1β、IL-6水平均显著高于PBS和GST蛋白对照组(P＜0.05)；血清中TNF-α、IL-1β水平显著高于PBS和GST蛋白对照组(P＜0.01).结论 pORF5蛋白能引起BALB/c小鼠生殖道组织免疫损伤,该损伤机制可能与TNF-α、IL-1β、IL-6等炎症因子水平升高有关.

abstractsObjective To investigate the immune injury in genital tract of BALB/c mice induced by plasmid protein pORF5 of Chlamydia trachomatis and its possible mechanism.Methods GST(glutathione-S-transferases)-pORF5 fusion protein was expressed and digested with PreScission Protease to obtain the target protein without GST tag.After further purification and endotoxin removal,pORF5 protein was injected into the posterior fornix of BALB/c mice on day 1,3 and 6,while the control groups were injected with PBS or GST protein respectively,and then all the mice were sacrificed on day 7 to evaluate genital tract gross pathology and histopathological characterization.The levels of TNF-α,IL-1β and IL-6 in serum,splenocytes culture supernatant and vaginal douche were detected by ELISA.Results Mice in pORF5 group developed different degrees of swelling in isthmic portion and ampulla of uterine tube,connective tissue adhesion and hydrosalpinx in the genital tract tissues,while the PBS group and the GST group did not show any obvious change.The inflammatory score showed that the genital tract pathology in pORF5 group was much more severe than PBS and GST control groups (P＜0.O1).The levels of TNF-α,IL-1β and IL-6 in vaginal douche and splenocytes culture supernatants in pORF5 group were obviously higher than those of PBS and GST groups (P＜0.05).The levels of TNF-α and IL-1β in serum were also higher than those of GST and PBS groups (P＜0.01).Conclusion pORF5 plasmid protein could induce pathological immune response in the genital tract of BALB/c mice,which may be associated with the increase of the production of the inflammatory cytokines TNF-α,IL-1β and IL-6 in BALB/c mice.