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| Study on anti-HBV effect of total phenolics acid from Oenanthe Javanica in vitro and its mechanism |
| LIU Qingchuan1 WANG Panli2 YANG Xinbo1 WANG Renjie1 WANG Mengmeng1 HUANG Zhengming1 |
1.Beijing Weijian Jiye Institute of Biotechnology, Beijing 100039, China;
2.Department of Pharmacy, People′s Hospital of Shigatse, Tibet Autonomous Region, Xigaze 857000, China |
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Abstract Objective To observe the inhibitory effect of total phenolics acid from Oenanthe Javanica on hepatitis B virus (HBV) in vitro and explore its mechanism. Methods The toxicity of total phenolics acid from Oenanthe Javanica to Hep AD38 cells was divided into the total phenolics acid from Oenanthe Javanica group (subdivided into 1600, 800, 400, 200, 100 μg/mL concentration groups) and the blank control group. Hep AD38 cells were cultured in vitro and the toxicity range of total phenolics acid from Oenanthe Javanica was determined by MTT assay. In vitro anti-HBV effects of total phenolics acid from Oenanthe Javanica were divided into the total phenolics acid from Oenanthe Javanica group (subdivided into 200, 100, 50 and 25 μg/mL concentration groups), the positive control group [Entecavir (ETV) 10 μmol/L] and the virus control group. Hep AD38 cells were treated with total phenolics acid from Oenanthe Javanica solution with a concentration below the non-toxic concentration for 6 consecutive days. The supernatant and cells were collected. The titer of HBsAg and HBeAg in the supernatant was detected by enzyme linked immune sorbent assay (ELISA). The intracellular HBV DNA load was detected by real-time fluorescent quantitative PCR. The effects of total phenolics acid from Oenanthe Javanica on duck hepatitis B virus (DHBV) DNA polymerase were divided into the total phenolics acid from Oenanthe Javanica group (subdivided into 400, 200, 100, 50 and 25 μg/mL concentration groups), the negative control group and the DHBV DNA polymerase control group. DHBV DNA polymerase was extracted from duck liver infected with duck hepatitis B virus, different concentrations of total phenolics acid from Oenanthe Javanica solution were given, and the effects of total phenolics acid from Oenanthe Javanica on DHBV DNA polymerase activity were detected by ELISA. Results The non-toxic concentration of total phenolics acid from Oenanthe Javanica in Hep AD38 cells was 400 μg/mL. Compared with the virus control group, HBsAg, HBeAg titer and HBV DNA load were significantly decreased in the 200, 100 and 50 μg/mL concentration groups of the total phenolics acid from Oenanthe Javanica group, the differences were highly statistically significant (P ＜ 0.01). Compared with the DHBV DNA polymerase control group, DHBV DNA polymerase activity was inhibited in the 400, 200, and 100 μg/mL concentration groups of the total phenolics acid from Oenanthe Javanica group, the differences were highly statistically significant (P ＜ 0.01). Conclusion Total phenolics acid from Oenanthe Javanicahas a good anti-HBV effect in vitro, and its mechanism includes inhibition of HBV DNA polymerase activity.
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