CCl4-induced liver fibrosis mouse model was also developed to investigate the role of TGR5 in liver fibrogenesis. Rosa26-LSL-Cas9 knockin mice (CAS9 KI, 8 weeks old) purchased from Jackson lab (https://www.jax.org/strain/024857 ) was divided into 4 groups (n = 8 in each group): (1) CAS9 KI+ corn oil group, ip with corn oil as vehicle; (2) CAS9 KI+CCl4 group, ip with 0.5 g/kg body weight CCl4 in corn oil (equivalent to 200 μL/mouse) 2 times a week for 5 weeks; (3) CAS9 KI+ CCl4+Tgr5 sgRNA group, the mice were treated with 100 µL adeno-associated virus (AAV) containing Gfap-Cre-U6-Tgr5 sgRNA cassette (virus concentration: 5.5E+10/mL) via intrasplenic injection once a week and ip with 0.5g/kg body weight CCl4 in corn oil (equivalent to 200 μL/mouse) 2 times a week for 5 weeks. The AAV can deplete Tgr5 expression specifically in HSCs due to the Gfap- promoter-driving- Cre recombinase expression. Twenty-four hours after the last challenge, all mice were sacrificed. All procedures were performed under sterile conditions. Serum and liver samples were collected for biochemical, histological and BA quantitation.
Animal welfare and the animal experimental protocols were strictly consistent with the Guide for the Care and Use of Laboratory Animals (U.S. National Research Council, 1996) and the related ethics regulations of Nanjing University Medical School.
Animal welfare and the animal experimental protocols were strictly consistent with the Guide for the Care and Use of Laboratory Animals (U.S. National Research Council, 1996) and the related ethics regulations of Nanjing University Medical School.