Journal: Cell reports

Article Title: Blockade of CCR5 suppresses paclitaxel-induced peripheral neuropathic pain caused by increased deoxycholic acid.

doi: 10.1016/j.celrep.2023.113386

Figure Lengend Snippet: Figure 6. DCA upregulates CCR5 expression through a TGR5/CREB-dependent mechanism (A) Reduced mechanical hyperalgesia in CCR5/ rats after DCA administration (n = 8). *p < 0.05 and **p < 0.01 by two-way ANOVA followed by Sida´ k’s multiple- comparison test. (B) TGR5 and FXR expressions in rat DRG by western blot (n = 4). (C and D) CCR5 mRNA levels in DRG neurons after siRNA knockdown of TGR5 and FXR in vitro (C) and with different DCA and TGR5 antagonist SBI-115 concentrations in vitro (D) (n = 3 cells). (E and F) Expression of TGR5 in sections of PIPN rat DRG. Double immunofluorescence images (E) and quantification analysis (F) detected the colocalization of TGR5 and markers of different population of DRG neurons. Scale bar, 50 mm. (G and H) Colocalization of TGR5 and CCR5 in PIPN rat DRG. Scale bar, 50 mm. (I–K) TGR5 knockdown in L4–L5 DRG neurons via shTGR5 AAV injections. (I) Experimental timeline. Knockdown efficacy confirmed by western blot (J) and immunofluorescence (K). n = 6 rats. Scale bar, 50 mm. (L and M) Protein expressions in rat DRG (n = 5). (N–R) Protein expressions in DRG neurons in vitro with DCA (N–O) and PKA- and PKC-selective inhibitors H89 and Go6983 (P) and transfected with siTGR5 (Q and R) in vitro (n = 3 or 4 cells).

Article Snippet: Following antibodies were used: CCR5 (1:100, Abcam#ab11466), IB4 (1:200, Invitrogen#I21411), CGRP (1:200, CST#14959; 1:200, Abcam#ab81887), NF200 (1:200, CST#2836), and TGR5 (1:200, Novus#NBP2-23669).

Techniques: Expressing, Comparison, Western Blot, Knockdown, In Vitro, Transfection

Journal: Cell reports

Article Title: Blockade of CCR5 suppresses paclitaxel-induced peripheral neuropathic pain caused by increased deoxycholic acid.

doi: 10.1016/j.celrep.2023.113386

Figure Lengend Snippet: Figure 7. Blockade of CCR5 by maraviroc or CCL5 neutralizing antibody improves recovery from PIPN (A) Acute treatment with maraviroc or CCL5 neuAb reverses established PIPN at 1 and 3 h after administration (n = 5). *p < 0.05, **p < 0.01, and ***p < 0.001, PIPN group vs. PIPN + maraviroc group; #p < 0.05, ##p < 0.01, and ###p < 0.001, PIPN group vs. PIPN + CCL5 neuAb group; p values by two-way ANOVA followed by Sida´ k’s multiple-comparison test. (B) Survival of 24-month-old rats during 14-day follow-up (n = 8). (C and D) Expression of CGRP in rat DRG after maraviroc and CCL5 neuAb treatment. Immunofluorescence images (C) and quantification analysis of the number of CGRP-IR neurons (D). Scale bar, 100 mm. *p < 0.05 and **p < 0.01 by one-way ANOVA with Tukey’s multiple-comparison post hoc test. (E) Expression of CCR5 and CCL5 in pathological classes of breast cancer patients (N = 1,097) and glioblastoma patients (N = 156) by UALCAN database. (F) Correlation of CCR5 and CCL5, TGR5, and FXR in tumor microenvironment by TIMER database. All data are presented as mean ± SEM.

Article Snippet: Following antibodies were used: CCR5 (1:100, Abcam#ab11466), IB4 (1:200, Invitrogen#I21411), CGRP (1:200, CST#14959; 1:200, Abcam#ab81887), NF200 (1:200, CST#2836), and TGR5 (1:200, Novus#NBP2-23669).

Techniques: Neutralizing Assay, Comparison, Expressing

Journal: Nutrients

Article Title: Glycochenodeoxycholate Affects Iron Homeostasis via Up-Regulating Hepcidin Expression

doi: 10.3390/nu14153176

Figure Lengend Snippet: Effects of GCDCA on the changes of BMP6/ALK3 signaling related receptors in HepG2 cells. ( A ) Relative gene expression of TGR5 and FXR after GCDCA treatment. ( B ) Representative western blotting bands of TGR5 and FXR under GCDCA treatment, as well as their fold changes in optical density relative to controls. ( C ) Relative gene expression of hepcidin and FXR after treatment of a FXR activator (GW4064, 10 µM for 2 h). ( D ) The effects of GW4064 on the phosphorylation of SMAD1/5/8 (normalized by total SMAD1) and protein level of FXR. ( E – H ) The effect of guggulsterone (GS, a FXR inhibitor, pretreated HepG2 cells for 18 h at 20 µM) on GCDCA-induced activation of FXR-BMP6/ALK3 signaling proteins and hepcidin expression. Values were shown as means ± SD. *, p < 0.05, GCDCA treatment group vs. control group. **, p < 0.01. ***, p < 0.001. †, p < 0.05, other experimental groups vs. control group. ††, p < 0.01. ##, p < 0.01, GCDCA treatment group vs. other experimental groups. ###, p < 0.001. Each experiment was performed at least three times.

Article Snippet: Primary antibodies rabbit anti-TGR5 (1:1000, NBP2-23669) and rabbit anti-ferroportin (1:1000, NBP1-21502) were purchased from Novus Biologicals (Littleton, Colorado, USA).

Techniques: Gene Expression, Western Blot, Phospho-proteomics, Activation Assay, Expressing, Control