Fig. 8: Blockage of macrophage S1PR2/ZBP1/p-MLKL alleviated necroptosis and further fibrosis in BDL liver.

A Mouse BMDMs were treated with 100 μmol/L sodium glycodeoxycholate (GDCA) for 6 h, with or without treatment of TSZ (TNFα plus Smac mimetic and z-VAD-FMK) for 6 h. Western blot analysis for ZBP1 and p-MLKL in mouse BMDMs. Blots of ZBP1 were normalized to GAPDH, and blots of p-MLKL were normalized to MLKL. B Lactate dehydrogenase (LDH) activities in cell supernatant of mouse BMDMs were measured in GDCA-, TSZ- or GDCA + TSZ-treated BMDMs. C Effects of Zbp1 siRNA on ZBP1 and p-MLKL protein expressions in mouse BMDMs treated with or without GDCA + TSZ. D The correlation between ZBP1 mRNA level and LSM (liver stiffness measurement) or liver fibrosis marker gene levels in BA patient and mouse model livers. E The mRNA expression of ZBP1 in BA liver tissues was compared between the jaundice-free and non-jaundice free groups after Kasai surgery at 6 months. F The schedule of mouse model. G mRNA expression of S1pr2 in liver tissue, non-parenchymal cells (NPCs) and macrophages from BDL mouse livers treated with glucan-encapsulated NC or S1pr2 siRNA particles (GeRPs). H, I Western blot analysis for ZBP1, p-MLKL, and Col I in the liver tissues from BDL mouse livers treated with NC (n = 3) or S1pr2 (n = 3) siRNA-GeRPs. Bolts of p-MLKL were normalized to MLKL, blots of ZBP1 and Col I were normalized to GAPDH. Data are presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 (versus control).