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目的建立土拨鼠慢性病毒性肝炎的动物模型。方法对一月龄的土拨鼠进行颈外静脉注射感染土拨鼠肝炎病毒(woodchuck hepatitis virus,WHV),使用两个剂量(高剂量:2.5×107 WID50;低剂量:5×106 WID50)感染,感染后在不同时间采用荧光定量PCR法测定血清中病毒DNA拷贝数,ELISA测定血清中WHsAg和WHcAb,并监测AST和ALT,肝脏活检HE染色观察病理变化,免疫组化检测WHsAg观察肝脏内病毒分布。结果感染后16周内,土拨鼠肝脏酶学AST和ALT水平均明显升高,土拨鼠血清内病毒载量在感染后第2周开始升高,16周达到高峰期,随后,低剂量组的病毒载量开始下降,而高剂量组的病毒载量仍接近106拷贝/ml。感染动物血清内可检测到WHsAg和WHcAb,肝脏病理切片显示小灶性肝细胞坏死伴炎细胞浸润及散在出血的急性肝炎症状,胞浆内可见WHsAg阳性分布。24周时,低剂量组土拨鼠血清内WHsAg和病毒DNA无法检出,提示其为急性自限性肝炎;高剂量组土拨鼠血清内依然可检测出WHsAg和病毒DNA,提示转为慢性感染。结论建立了急性病毒性肝炎转为慢性感染的土拨鼠模型,提示急性肝炎的病程发展除与土拨鼠感染年龄相关外,也与病毒感染剂量相关。
Objective To establish an animal model of woodchuck chronic viral hepatitis. Methods One-month-old groundhogs were infected with woodchuck hepatitis virus (WHV) by external jugular vein injection using two doses (high dose: 2.5 × 107 WID50; low dose: 5 × 106 WID50) , At different times after infection, the virus DNA copy number in serum was determined by fluorescence quantitative PCR, WHsAg and WHcAb in serum were measured by ELISA, AST and ALT were detected, liver biopsy HE staining to observe the pathological changes, immunohistochemical detection WHsAg liver virus distributed. Results Within 16 weeks after infection, the levels of AST and ALT in liver of marmots were significantly increased. The serum viral load in marmots began to increase from the second week after infection and peaked at 16 weeks. Subsequently, the low dose The viral load of the group started to decline, while the viral load of the high dose group was still close to 106 copies / ml. WHsAg and WHcAb were detected in the serum of infected animals. The pathological findings of hepatic necrosis with inflammatory cell infiltration and scattered hemorrhage in acute hepatic symptoms showed a positive distribution of WHsAg in cytoplasm. At 24 weeks, WHsAg and viral DNA were undetectable in the serum of low-dose group of woodchuck mice, suggesting that they were acute, self-limiting hepatitis; WHsAg and viral DNA were still detectable in the serum of high-dose group of woodchuck dogs, suggesting a shift to chronic infection. Conclusion The establishment of a woodchuck model of acute viral hepatitis into chronic infection, suggesting that the course of acute hepatitis in addition to the age of marmots associated with age, but also with the dose of viral infection.