OBJECTIVETo observe the effect of Shugan Jianpi Bushen Recipe (SJBR) on the splenic T lymphocytes and virus load in the hepatitis B virus (HBV) transgenic (Tg) mice, and to study its antiviral efficacy and mechanisms of action.

METHODSSixty male BALB/C mice of SPF grade were included. Ten non-HBV Tg male mice were included as the normal control group. Fifty HBV Tg mice were randomly divided into five groups, i. e., the model group, the adefovir (ADV) group, the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group, ten in each. 10, 20, and 40 g/kg SJBR crude drug was respectively given by gastro-gavage to mice in the low dose SJBR group, the middle dose SJBR group, and the high dose SJBR group. ADV 50 mg/kg body weight was given by gastrogavage to mice in the ADV group. Equal volume of sterilized iso-osmia was given to mice in the normal group and the model group. The medication was performed once daily, totally for 21 successive days. The serum HBV DNA titers of HBV Tg mice were detected using Real-time fluorescent PCR one day before administration (T0), ten days after administration (T1), 21 days after administration (T2), and three days after withdrawal (T3), respectively; the serum hepatitis B surface antigen (HBsAg) of HBV Tg mice on T3 was detected by ELISA. The splenic T lymphocyte percent of all mice was detected by flow cytometry.

RESULTSSerum HBsAg at TO was positive in the high-, middle-, low-dose SJBR, and ADV groups. The HBsAg negative rate at T3 was lower in the high dose SJBR group than in the ADV group, showing statistical difference (P<0.01). Compared with TO of the same group, the serum HBV DNA titers could be continually decreased by high dose SJBR, showing statistical difference (P<0.01). The serum HBV DNA titers also gradually decreased in the ADV group (P<0.01), but it somewhat increased at T3. The CD3+ cell percent could be elevated by high-, middle-, low-dose SJBR, and ADV groups (P<0.05, P<0.01). The CD8+ T cell percent could also be obviously lowered by high-, middle-, and low-dose SJBR (P<0.01). Compared with the middle-, low-dose SJBR, and ADV groups, the CD4+ T cell percent and CD4+/CD8+ increased as well as CD8+ decreased in the high dose SJBR group, showing statistical difference (P<0.01). The CD3+ T cell percent was significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the middle dose SJBR group (r=0.654, P<0.05). The percents of CD4+, CD8+ T cells and CD4+/CD8+ were significantly positively correlated to the decrement of HBV DNA titers between the pre-treatment and post-treatment in the high dose SJBR group (r=0.53, r=0.79, r =0.80, P<0.01).

CONCLUSIONSSJBR were capable of inhibiting the HBV DNA duplication of HBV Tg mice. One of its anti-HBV mechanisms possibly be improving the the abnormality of T lymphocyte subsets and the immune function.