To investigate the effect of miR‐188‐5p inhibition on HF in vivo, we used diet and chemical‐induced dual fibrosis model. In several studies, mild dose of intraperitoneal (i.p.) CCl4 injections to animals fed on HFD has been accepted as a promised and rapid liver fibrosis model by triggering inflammation, steatosis and fibrosis which are histological features of NASH. 19 , 20 Therefore, using the same strategy, 40 male C57BL/6J mice were randomized into five groups (n = 8 mice in each group): Control group, mice were fed on normal diet; HFD group, mice were fed on HFD (Research Diet, D12492) along with weekly 200 μl i.p. injection of corn oil (Shanghai Aladdin Biochemical, C116025) for 14 weeks; HFD + CCl4 group, mice were fed with HFD along with weekly i.p. administration of CCL4 (Tianli Chemical Reagent, GB/T688‐2011) diluted in corn oil (CCl4 at dose rate of 0.25 μl (0.40 μg)/g of body weight diluted in 200 μl of corn oil) for 14 weeks; HFD + CCl4‐miR‐NC group; and HFD + CCl4‐miR‐188‐5p‐inhibitor group. To observe the anti‐fibrotic effect of miR‐188‐5p inhibition, 62.5 nM/kg dose of miR‐188 inhibitors or miR‐NC complexed with Lipofectamine® 3000 (Invitrogen, L3000015) were diluted in 50 μl of sterile normal saline and injected in respective groups of mice through tail vein injection at the mid of 5th week, start of 6th and 7th week. The schematic representation of animal model groups used in the study for the inhibition of miR‐188‐5p is presented in Figure S2. At the 14th week of experiment, all mice were sacrificed. Liver tissues were collected and divided into two parts. One part was immediately snap‐frozen in liquid nitrogen and then stored at −80°C for further experiments, while another section was fixed in formalin and embedded in paraffin, for histological analysis.

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