Background and Purpose:  Macrophages are central immune characters in hepatic fibrosis by reconstructing the fibrotic immune microenvironment. Picroside II (PIC II) has exerted a therapeutic potential on liver injury. However, the mechanisms by which macrophage initiates immune cascades and further contributes to liver fibrosis and whether this process can be influenced by PIC II remains unclear. Experimental Approach: In this research, the RNA sequencing of multidrug-resistance protein 2 knockout (Mdr2-/-) mice was applied. Then aHSCs were incubated with the medium from M1 macrophages and NK cells, with the extra formation of neutrophils extracellular traps (NETs) being tested. In addition, we intraperitoneal injected PIC II and then intravenously injected the clodronate liposome to evaluate the therapeutic effect of PIC II and macrophage deletion in Mdr2-/- mice. Key Results: We observed the increase of CXCL16+ M1 macrophages in Mdr2-/- liver, accompanied by the recruitment of CXCR6+ NK cells and NETs formation. PIC II promoted the CXCL16+ macrophage recruited NK cells via CXCL16/CXCR6 axis, which subsequently affecting the JAK1/STAT1 signaling in aHSCs. And fibrotic liver was relieved to some extent when PIC II combined with macrophage depletion. Conclusion and Implications: Mechanistically, PIC II activated M1 macrophage to recruit NK cells via CXCL16-CXCR6 axis and subsequently regulated the JAK1/STAT1 signaling to restrain aHSCs. PIC II also alleviated fibrosis by attenuating the formation of NETs. Notably, these PIC II-associated hepatoprotective effects were largely reversed by macrophage depletion in Mdr2-/- mice. Collectively, our research suggests that PIC II is potential for halting the liver fibrosis.