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目的研究TLR2信号对华支睾吸虫易感性和成虫发育的影响。方法 TLR2野生型和变异型C57BL/6J小鼠灌胃感染60个华支睾吸虫囊蚴,同时设立未感染对照组。小鼠感染后第10d开始连续收集粪便;于感染后30、60、90和120d解剖小鼠,观察肝胆管病变,同时收集血清并制作肝脏组织标本。用ELISA法检测血清华支睾吸虫IgG相对水平;用Kato-Katz法检查小鼠粪便中的华支睾吸虫虫卵;肝脏组织经固定、切片后进行HE染色,显微镜下观察病理变化。结果 TLR2野生型和变异型C57BL/6J小鼠感染华支睾吸虫后血清特异IgG水平升高,且感染30d时的A450值后者高于前者(t=3.543,P<0.05);感染小鼠粪便内均未检出虫卵;感染小鼠的胆总管扩张,以TLR2变异型小鼠尤其明显;TLR2变异型小鼠感染60d时肝内胆管见有华支睾吸虫幼虫,TLR2野生型小鼠肝内胆管未检出华支睾吸虫;TLR2变异型小鼠感染120d时镜检肝细胞片状坏死,胆管组织恶性增生,TLR2野生型小鼠未发生上述病变。结论TLR2变异型C57BL/6J小鼠对华支睾吸虫的易感性增高。TLR2信号是C57BL/6J小鼠抵抗华支睾吸虫感染的重要因素,并对虫体发育和肝胆管组织恶性转化发挥抑制作用。
Objective To study the effect of TLR2 signaling on the susceptibility and adult development of Clonorchis sinensis. Methods Totally 60 CFU of Clonorchis sinensis were inoculated intragastrically in TLR2 wild-type and mutant C57BL / 6J mice. At the same time, uninfected control group was established. Mice were continuously infected with faeces 10 days after infection. Mice were dissected at 30, 60, 90 and 120 days after infection to observe hepatobiliary diseases. Meanwhile, serum was collected and liver samples were prepared. The relative level of serum IgG was detected by ELISA. Clonorchis sinensis eggs were detected by Kato-Katz method. The liver tissues were fixed and sliced for HE staining. The pathological changes were observed under the microscope. Results The level of serum specific IgG increased after infected with Clonorchis sinensis in both TLR2 wild type and mutant C57BL / 6J mice, and the latter was higher than the former at the 30th day after infection (t = 3.543, P <0.05) Infected mice were not detected in eggs; infected mice common bile duct dilatation, TLR2 mutant mice in particular; TLR2 mutant mice infected with intrahepatic bile duct see clonorchiasis larvae, TLR2 wild-type mice Clonorchis sinensis was not detected in the intrahepatic bile duct; TLR2 mutant mice microscopically detected hepatocellular plaque necrosis and bile duct malignant hyperplasia at 120 days after infection. TLR2 wild type mice did not develop these lesions. Conclusion The susceptibility of Clonorchis sinensis to TLR2 variant C57BL / 6J mice increased. TLR2 signal is an important factor in C57BL / 6J mice resistant to Clonorchis sinensis infection, and inhibit the development of worms and the malignant transformation of hepatobiliary tissue.