Fig. 4

Asiatic acid inhibited HBV replication and transcription. (A-B) HBV-infected HepG2-NTCP cells were treated with asiatic acid(10, 20 and 30 µM) or ETV for 3, 6, 9 days. Total HBV RNAs and HBV 3.5-kb RNA were detected by real-time PCR; (C) HBV 3.5-kb RNA and 2.4/2.1-kb RNA were examined by Northern blotting, ribosomal RNAs (28 S and 18 S) were served as a loading control; (D,E) The level of HBV core DNA was analyzed by real-time PCR and Southern blot; (F) ELISA was performed to detect the HBsAg level in supernatant of HBV-infected HepG2-NTCP cells; (G-J) PHHs were treated with asiatic acid(10, 20, 30µM) or ETV for 4 days. HBV cccDNA was detected by Taq-man probe PCR. Total HBV RNAs and HBV 3.5-kb RNA were quantified by real-time PCR. The ratios of total HBV RNAs/cccDNA and HBV 3.5-kb RNA/cccDNA were calculated as indicators of cccDNA transcription activity; (K,L) The level of HBV core DNA was analyzed by real-time PCR analysis and HBsAg in supernatant of HBV-infected HepG2-NTCP cells was subjected to ELISA. Representative data are from at least three independent experiments. Data are shown as mean ± SD, *P < 0.05; **P < 0.01