Diagnosis and Treatment of Infectious Diseases Research Laboratory, Shenzhen Third People's Hospital, Shenzhen, 518112, China.
Anhui Academy of Medical Sciences, Hefei, 230061, China.
Inflamm Res. 2019 Dec;68(12):1025-1034. doi: 10.1007/s00011-019-01284-2. Epub 2019 Sep 17.
Saikosaponin c (SSc), a compound purified from the traditional Chinese herb of Radix Bupleuri was previously identified to exhibit anti-HBV replication activity. However, the mechanism through which SSc acts against HBV remains unknown. In this study, we investigated the mechanism of SSc mediated anti-HBV activity.
HepG2.2.15 cells were cultured at 37 ℃ in the presence of 1-40 μg/mL of SSc or DMSO as a control. The expression profile of HBV markers, cytokines, HNF1α and HNF4α were investigated by real-time quantitative PCR, Elisa, Western blot and Dot blotting. Knockdown of HNF1α or HNF4α in HepG2.2.15 cells was mediated by two small siRNAs specifically targeting HNF1α or HNF4α.
We found that SSc stimulates IL-6 expression, leading to attenuated HNF1α and HNF4α expression, which further mediates suppression of HBV pgRNA synthesis. Knockdown of HNF1α or HNF4α in HepG2.2.15 cells by RNA interference abrogates SSc's anti-HBV role. Moreover, SSc is effective to both wild-type and drug-resistant HBV mutants.
SSc inhibits pgRNA synthesis by targeting HNF1α and HNF4α. These results indicate that SSc acts as a promising compound for modulating pgRNA transcription in the therapeutic strategies against HBV infection.
柴胡皂甙 c（SSc）是从传统中药柴胡中分离得到的一种化合物，先前被鉴定具有抗 HBV 复制活性。然而，SSc 作用于 HBV 的机制尚不清楚。在本研究中，我们研究了 SSc 介导的抗 HBV 活性的机制。
在 37℃下，用 1-40μg/mL 的 SSc 或 DMSO（对照）培养 HepG2.2.15 细胞。通过实时定量 PCR、Elisa、Western blot 和 Dot blotting 研究 HBV 标志物、细胞因子、HNF1α 和 HNF4α 的表达谱。通过两种针对 HNF1α 或 HNF4α 的特异性小 siRNA 敲低 HepG2.2.15 细胞中的 HNF1α 或 HNF4α。
我们发现 SSc 刺激 IL-6 表达，导致 HNF1α 和 HNF4α 表达减弱，从而介导抑制 HBV pgRNA 合成。用 RNA 干扰敲低 HepG2.2.15 细胞中的 HNF1α 或 HNF4α 可消除 SSc 的抗 HBV 作用。此外，SSc 对野生型和耐药 HBV 突变体均有效。
SSc 通过靶向 HNF1α 和 HNF4α 抑制 pgRNA 合成。这些结果表明，SSc 作为一种有前途的化合物，可用于调节 HBV 感染治疗策略中的 pgRNA 转录。
