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目的:建立鸭乙型肝炎慢性感染模型,研究玉郎伞水提物对鸭慢性乙型肝炎的保护作用。方法:1日龄的广西麻鸭经颈静脉接种0.2 m L的鸭乙型肝炎病毒(DHBV)强阳性血清,7 d后用荧光定量PCR筛选DHBV阳性鸭,随机分为4组,即:模型组,玉郎伞水提物高、低剂量组(按生药量计分别为20,10 g·kg-1)和秋水仙碱组(0.02 g·kg-1)。将未感染鸭作为正常组。除正常组外,其余各组每周均接种0.2 m L的DHBV强阳性血清,直到实验结束,以造成鸭乙型肝炎慢性感染。从造模第25周开始,连续灌胃给药5周。末次给药24 h后,所有动物从颈静脉采血,测定血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性,血清鸭乙型肝炎病毒表面抗原(DHBs Ag),DHBV-DNA含量。将所有动物处死,迅速摘取肝组织,测定肝匀浆超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px)的活性,丙二醛(MDA)的含量,肝组织中羟脯氨酸(Hyp)的含量。通过HE染色观察肝细胞损伤程度。结果:与正常组比较,模型组血清ALT,AST活性,HBs Ag,DHBV-DNA含量均显著升高(P<0.05,P<0.01)。肝组织中SOD,GSP-Px活性均显著降低,MDA和Hyp含量则显著升高(P<0.05,P<0.01)。与模型组比较,玉郎伞水提物高、低剂量组均能降低血清ALT活性和DHBV-DNA含量(P<0.05,P<0.01),均能增加肝组织匀浆中GSH-Px活性,降低MDA和Hyp的含量(P<0.05,P<0.01)。玉郎伞水提物高剂量组能降低血清AST活性和HBs Ag含量,增加肝组织SOD活性(P<0.05,P<0.01)。HE染色显示玉郎伞水提物能显著减轻肝细胞损伤。结论:玉郎伞水提物对DHBV所致的慢性鸭乙型肝炎有一定的保护作用。
Objective: To establish a chronic infection model of duck hepatitis B and to study the protective effect of the aqueous extract of Ylang Lang on chronic hepatitis B in duck. Methods: One-day-old Guangxi ducklings were inoculated with 0.2 mL DHBV positive sera via the jugular vein. DHBV-positive ducks were screened by fluorescence quantitative PCR 7 days later, and were randomly divided into 4 groups: model Group, the high and low dose of Yulangsan water extract (20 and 10 g · kg-1, respectively) and the colchicine group (0.02 g · kg-1). Uninfected ducks as normal group. Except for the normal group, all the other groups were inoculated with 0.2 m L DHBV positive serum every week until the experiment was over to cause chronic hepatitis B infection. Starting from the 25th week of model establishment, continuous gavage was given for 5 weeks. After the last administration for 24 h, all the animals were taken blood from the jugular vein to measure the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), serum duck hepatitis B virus surface antigen (DHBs Ag), DHBV-DNA content. All animals were sacrificed and the liver tissue was rapidly removed. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), the content of malondialdehyde (MDA) Hyp in tissues. Hepatic injury was observed by HE staining. Results: Compared with the normal group, the levels of ALT, AST, HBsAg and DHBV-DNA in model group were significantly increased (P <0.05, P <0.01). The activities of SOD and GSP-Px in liver tissue were significantly decreased, MDA and Hyp contents were significantly increased (P <0.05, P <0.01). Compared with the model group, the ALT activity and DHBV-DNA content in the serum of both groups were significantly decreased (P <0.05, P <0.01), both of which could increase the activity of GSH-Px in the liver homogenate, Reduce the content of MDA and Hyp (P <0.05, P <0.01). High dose Yulangsan water extract group can reduce serum AST activity and HBsAg content, increase SOD activity in liver tissue (P <0.05, P <0.01). HE staining showed Yulangsan water extract can significantly reduce liver cell damage. Conclusion: The water extract of Ylang Lang has certain protective effect on DHBV induced chronic duck hepatitis B.