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目的:观察玉郎伞多糖(YLSP)在鸭乙型肝炎病毒(DHBV)持续性感染模型中对鸭血清乙型肝炎病毒DNA的抑制作用及保护肝细胞的作用。方法:将DHBV-DNA强阳性麻鸭随机分为YLSP高、中、低剂量组(10,5,2.5 g·kg-1)、拉米夫定(3TC,0.05 g·kg-1)组和模型组,以上各组每日上午灌胃给药l次,连续14 d。于用药前(T0)、用药7 d(T7)和14 d(T14)及停药后3 d(P3)取静脉血用实时荧光定量PCR检测DHBV-DNA含量,同时采用酶联免疫吸附(ELISA)法测定上清液鸭乙型肝炎病毒表面抗原(DHBsAg)和e抗原(DHBeAg)的滴度;在停药3 d后,取肝脏匀浆,检测肝匀浆液中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性以及丙二醛(MDA)、谷胱甘肽(GSH)的含量。结果:与模型组相比,YLSP高、中剂量治疗组给药7 d(T7)和14 d(T14)即能明显抑制DHBV-DNA复制及血清DHBsAg,DHBeAg的滴度显著降低(P<0.05或P<0.01),停药后3 d(P3),YLSP高剂量组仍能显示出持续有效,血清DHBV-DNA水平及血清DHBsAg,DHBeAg的滴度均无反跳现象(P<0.05或P<0.01);在停药3d后,肝匀浆液中SOD,GSH-Px活性以及MDA,GSH的含量,YLSP高、中剂量组仍能显示出持续有效,没有出现反跳现象(P<0.05或P<0.01)。结论:YLSP具有有效抑制DHBV-DNA和保护肝细胞的作用。
Objective: To observe the inhibitory effect of Yulangsan polysaccharide (YLSP) on duck hepatitis B virus DNA and the protection of hepatocytes in Duck Hepatitis B virus (DHBV) persistent infection model. Methods: DHBV-DNA strongly positive ducks were randomly divided into three groups: YLSP high, middle and low dose groups (10,5,2.5 g · kg -1), lamivudine group (3TC, 0.05 g · kg -1) Model group, the above groups were administered orally once a day for 14 days. The levels of DHBV-DNA were detected by real-time fluorescence quantitative PCR before 7 days (T7), 14 days (T14) and 3 days after drug withdrawal (P3). Meanwhile, the enzyme linked immunosorbent assay ) Method was used to determine the titer of DHBsAg and DHBeAg in the supernatant. After 3 days of withdrawal, the liver homogenates were taken for the detection of superoxide dismutase (SOD) , Glutathione peroxidase (GSH-Px) activity and malondialdehyde (MDA), glutathione (GSH) content. Results: Compared with the model group, the titer of DHBAg and DHBV in DHBV-DNA replication and serum DHBAg were significantly decreased in the high-dose and middle-dose YTSP treated groups for 7 d (T7) and 14 d (T14) (P <0.05 or P <0.01, P <0.01 or P <0.01). After 3 days (P3), the YLSP high dose group continued to be effective and the serum DHBV-DNA levels and serum DHBsAg and DHBeAg titres did not rebound <0.01). After three days’ withdrawal, the activities of SOD, GSH-Px and MDA and GSH in the liver homogenate and the YLSP high and middle dose groups still showed no signs of rebound (P <0.05 or P <0.01). Conclusion: YLSP can effectively inhibit DHBV-DNA and protect liver cells.