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目的应用聚肌胞苷酸(polyinosinic polycytidylic acid,poly I:C)建立原发性胆汁性肝硬化(PBC)动物模型。方法80只C57BL/6雌性小鼠,分为空白对照组(5只)、poly I:C 5mg/kg组(25只)和10mg/kg组(25只)、阴性对照组(25只)。poly I:C 5和10mg/kg组小鼠腹腔分别注射poly I:C 5和10mg/kg,每3天1次,阴性对照组用等体积无菌PBS液注射。每4周处死一批小鼠,同时检测血清丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP)、抗核抗体(ANA)、抗线粒体抗体(AMA)水平,肝组织行H-E染色后镜下观察胆管组织学变化;Annexin V检测脾细胞凋亡。结果5mg/kg组第4周小叶间胆管开始出现炎性细胞浸润,浸润的胆管数量随注射时间延长而增加,第20周达51%;血清AMA阳性率逐渐增高,16周达4只(共5只),最高滴度达1:10000。10mg/kg组ANA、AMA均在第8周时明显升高,但随后呈下降趋势,其他指标与5mg/kg组差异无统计学意义(P>0.05)。5mg/kg组血清ALP显著高于阴性对照组(P<0.05)。poly I:C两组脾细胞早期凋亡率均明显高于阴性对照组(P<0.05),但两组间差异无统计学意义(P>0.05)。阴性对照组仅出现ANA阳性,其他指标均无明显变化。结论poly I:C(5mg/kg)注射16周后可初步建立PBC动物模型。细胞凋亡后释放的自身抗原活化自身反应性淋巴细胞在PBC的发病中可能起重要作用。该模型的建立为今后PBC发病机制和治疗的研究奠定了基础。
Objective To establish a primary animal model of biliary cirrhosis (PBC) using poly-I (C) poly I (C). Methods Eighty C57BL / 6 female mice were divided into blank control group (n = 5), poly I: C 5 mg / kg group (n = 25) and 10 mg / kg group (n = 25). Negative control group (n = 25). The mice in the poly I: C 5 and 10 mg / kg groups were injected intraperitoneally with poly I: C 5 and 10 mg / kg once every 3 days. The negative control group was injected with an equal volume of sterile PBS solution. A group of mice were sacrificed every 4 weeks, serum ALT, ALP, ANA and AMA levels were also measured. The liver tissues were examined with HE staining Under the observation of histological changes of bile duct; Annexin V apoptosis of spleen cells. Results Inflammatory cell infiltration was observed in the interlobular septae of 5 mg / kg group at 4 weeks. The number of infiltrating bile duct increased with the time of injection, reaching 51% in the 20 th week. The positive rate of serum AMA gradually increased and reached 16 at 16 weeks 5), the highest titer reached 1: 10000.10mg / kg group ANA, AMA were significantly increased at the 8th week, but then decreased, other indicators and 5mg / kg group no significant difference (P> 0.05). Serum ALP in 5mg / kg group was significantly higher than that in negative control group (P <0.05). The early apoptotic rate of splenocytes in both poly I: C groups was significantly higher than that in the negative control group (P <0.05), but there was no significant difference between the two groups (P> 0.05). Negative control group only appeared ANA positive, no significant change in other indicators. Conclusion PBC animal model can be initially established after poly I: C (5mg / kg) injection for 16 weeks. Autologous antigen released after apoptosis Activation of autoreactive lymphocytes may play an important role in the pathogenesis of PBC. The establishment of this model lays the foundation for the research on the pathogenesis and treatment of PBC in the future.