Additional file 1 of Tricyclic antidepressants induce liver inflammation by targeting NLRP3 inflammasome activation
Creators: name: Mu, Wenqing
name: Xu, Guang
name: Wang, Zhilei
name: Li, Qiang
name: Sun, Siqiao
name: Qin, Qin
name: Li, Zhiyong
name: Shi, Wei
name: Dai, Wenzhang
name: Zhan, Xiaoyan
name: Wang, Jiabo
name: Bai, Zhaofang
name: Xiao, Xiaohe
Publisher: name: figshare
Year: 2023
Subjects: subject: Medicine
subject: Cell Biology
subject: Physiology
subject: FOS: Biological sciences
subject: Pharmacology
subject: Environmental Sciences not elsewhere classified
subject: Chemical Sciences not elsewhere classified
subject: Science Policy
subject: Immunology
subject: FOS: Clinical medicine
subject: Cancer

description: Additional file 1: Supplementary information on liver injury induced by TCAs by triggering NLRP3 inflammasome activation. Fig. S1. Cell viability of BMDMs administrated with nortriptyline was assessed using CellTiter-Glo Assay, which is according to quantitation of ATP. Fig. S2. Tricyclic antidepressantnortriptyline triggers the inflammasome activation in the absence of agonists.LPS-primed BMDMs were treated with carbamazepine and nortriptyline for 12 h or treated with them for 1 h followed by ATP stimulation, respectively. Western blotting was used to assess the expression of caspase-1 and IL-1β in SN. Fig. S3. Nortriptyline has no effect on Ca2+ mobilization.LPS-primed BMDMs were pretreated with EDTAand then stimulated with nortriptyline. Western blotting was used to assess the expression of caspase-1 in SN. Fig. S4. Nortriptyline activates NLRP3 inflammasome by inducing the accumulation of mtROS.LPS-primed BMDMs were pretreated with NACfor 1 h and then treated with nortriptyline for 6 h. The content of mtROSwas measured by flow cytometry. Western blotting was used to assess the expression of IL-1β and caspase-1 in cell SN as well as pro-IL-1β, pro-caspase-1, and NLRP3in WCL. The level of caspase-1activity. The levels of IL-1βand TNF-αin SN using ELISA. Data are presented as mean ± SEM; *P &lt; 0.05, ***P &lt; 0.001; One-Way ANOVA followed by Dunnett's post hoc test. Fig. S5. Multiple TCAs specifically trigger the aberrant activation of NLRP3 inflammasome.LPS-primed BMDMs were pretreated with MCC950 and then stimulated with Imi, Ami, Pro, and Nor for 12 h, the levels of IL-1βand TNF-αwere detected by ELISA kits.WT and Nlrp3-/- BMDMs were primed with LPS and then stimulated with Imi, Ami, Pro and Nor, the levels of IL-1βand TNF-αwere evaluated by ELISA kits. Data are expressed as the mean ± SEM,; *P &lt; 0.05, **P &lt; 0.01, ***P &lt; 0.001 vs. the control; ns, not significant; unpaired Student’s t-testfollowed by the Dunnett's post hoc test. Fig. S6. Multiple TCAs induce the NLRP3 inflammasome activation by triggering upstream signaling events.LPS-primed BMDMs were pretreated with MCC950, NAC, and KCl for 1 h followed by these TCAs stimulation, respectively. Western blotting was used to assess the expression of caspase-1in SN. Fig. S7. Nortriptyline directly triggered activation of the NLRP3 inflammasome in both hepatocytes and Kupffer cells.LPS-primed BMDMs and hepatocytes were pretreated with MCC950 and then stimulated with nortriptyline for 12 h. The levels of IL-1β in SN were detected using ELISA kits.BMDMs and Kupffer cells were primed with LPS and then pretreated with MCC950 followed by nortriptyline stimulation for 12 h. The levels of IL-1β in SN were detected using ELISA kits. Data are presented as the mean ± SEM; ***P &lt; 0.001; One-Way ANOVA followed by the Dunnett's post hoc test. Fig. S8. Nortriptyline induces IDILI by triggering the activation of NLRP3 inflammasome.Female WT C57BL/6 mice were pretreated with LPSand then treated with different doses of nortriptyline. The serum levels of TNF-α were detected by ELISA kits.WT or Nlrp3-/- mice were pretreated with LPS and then treated with nortriptyline. ELISA kits were used to determine the levels of IL-6and TNF-αin mouse sera. Data are shown as the mean ± SEM; ***P &lt; 0.001; ns, not significant. Statistics differences were analyzed using an unpaired Student’s t-test.