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摘要：
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摘要：目的 采用体外细胞模型评价保肝护肝药物舒肝宁注射液（Shuganning Zhusheye，SGN）对
HepG2.2.15细胞中稳定复制的野生型（wild type，WT）HBV和HepG2.A64细胞中稳定复制的恩替卡韦
（entecavir，ETV）耐药型HBV的抗病毒及协同抗病毒作用。方法 采用CCK8细胞计数法评价SGN细
胞毒性作用，选择安全用药浓度系列稀释后单独或与对照药物 [ETV及替诺福韦酯（tenofovir disoproxil
fumarate，TDF）] 联合进行抗HBV实验，以对HBV DNA、HBsAg和HBeAg的抑制率评价SGN的抗病
毒作用。结果 SGN对HepG2.2.15和HepG2.A64细胞的CC 50 分别为198.40 μg/ml和85.81 μg/ml；SGN单独
使用对HepG2.2.15和HepG2.A64细胞中HBV DNA的最大抑制率分别为55.89%（t = 14.195，P = 0.0094）
和46.29%（t = 4.953, P = 0.0037），SGN联合ETV对HepG2.2.15和HepG2.A64细胞中HBV DNA的最大抑
制率分别为94.28%（t = 9.745，P = 0.00068）和54.75%（t = 9.335，P = 0.00028），SGN联合TDF组对
HepG2.2.15和HepG2.A64细胞中HBV DNA的最大抑制率分别为93.11%（t = 9.274，P = 0.00071）和
88.27%（t = 43.68，P = 6.03 × 10 -6 ）；SGN对野生型HBsAg和HBeAg未见明显抑制作用（抑制率＜
12.00%），对ETV耐药型HBsAg抑制率为19.69%～34.26%（t = 39.118，P = 0.028；t = 19.73，P =
0.0024）。8 μg/ml SGN与低浓度（0.01 μmol/L）ETV联合对WT型HBV DNA的抑制有协同效应，抑制率
为86.28%（t = 30.745，P = 0.001）。8 μg/ml SGN与低浓度（0.2 μmol/L）TDF联合对ETV耐药型HBV的抑
制有协同效应，抑制率为60.26%（t = 7.568，P = 0.017）。结论 SGN可显著抑制WT型和ETV耐药型HBV
的复制，联合低浓度ETV或TDF有显著协同抑制作用。
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Abstract: Objective To evaluate the antiviral and synergistic antiviral effects of Shuganning Zhusheye
(SGN) on wild-type (WT) HBV and entecavir (ETV)-resistant HBV in HepG2.2.15 cell models and HepG2.
A64 cell models in vitro, respectively. Methods Cell counting kit-8 (CCK8) method was used to evaluate
the cytotoxicity of SGN, and the cell-safe serial-diluted concentration of SGN single or combined with ETV
and tenofovir disoproxil fumarate (TDF) were selected for antiviral assay. The antiviral effects of SGN were
evaluated with the inhibition rates of HBV DNA, HBsAg, and HBeAg levels. Results The CC 50 of SGN in
HepG2.2.15 cells and HepG2.A64 cells were 198.40 μg/ml and 85.81 μg/ml, respectively. The maximum
inhibition rate of HBV DNA was 55.89% (t = 14.195, P = 0.0094) and 46.29% (t = 4.953, P = 0.0037) in
SGN group, 94.28% (t = 9.745, P = 0.00068) and 54.75% (t = 9.335, P = 0.00028) in SGN combined ETV
group, and 93.11% (t = 9.274, P = 0.00071) and 88.27% (t = 43.68, P = 6.03 × 10 -6 ) in SGN combined
TDF group, respectively. However, SGN had no significant inhibition effect on HBsAg and HBeAg
levels in HepG2.2.15 cells (inhibition rate＜ 12.00%). The inhibition rate of HBsAg in SGN group was
19.69%～34.26% (t = 39.118, P = 0.028; t = 19.73, P = 0.0024) in HepG2.A64 cells. SGN (8 μg/ml) combined
with low concentration (0.01 μmol/L) of ETV had a synergistic antiviral effect on WT HBV DNA and the
inhibition rate was 86.28% (t = 30.745, P = 0.001). SGN (8 μg/ml) combined with low concentration (0.2
μmol/L) of TDF had a synergistic antiviral effect on ETV-resistant HBV and the inhibition rate was 60.26%
(t = 7.568, P = 0.017). Conclusions SGN can effectively inhibit the replication of both WT and ETV-
resistant HBV, and combination of SGN with low concentrations of ETV or TDF also have a significantly
synergistic inhibitive efficacy.
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