Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: Expression of PTP1B and double immunofluorescence of PTP1B, and NeuN, Iba‐1, and GFAP in the ipsilateral spinal dorsal horn of sham and neuropathic pain rats. (A) Mechanical allodynia was evaluated by the paw withdrawal threshold (PWT) at baseline and 3, 7, and 14 days after surgery. There is no significant difference regarding the PWT among the sham and SNI groups at baseline. However, the PWT in SNI rats was markedly decreased from day 3 to day 14 (* p < 0.05, **** p < 0.0001 compared with sham group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests). (B) Western blot results showed that the protein level of PTP1B in the spinal cord of rats with SNI was markedly increased from day 3 to day 14 (**** p < 0.0001, n = 6 rats per group, one‐way ANOVA followed by Bonferroni post‐hoc test). (C) Double immunofluorescence of PTP1B and NeuN, Iba‐1, and GFAP in the ipsilateral spinal dorsal horn of sham and SNI rats.

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques: Expressing, Immunofluorescence, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: Analgesic effect of PTP1B siRNA or PTP1B‐IN‐1 on mechanical allodynia in neuropathic pain rats. (A) In vitro study showed that the expression of PTP1B protein in cultured HEK293T cells was suppressed by the treatment of PTP1B siRNA. ** p < 0.01 versus control or scramble (sc) RNA, n = 3 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test. (B) Western blot assay showed that SNI‐induced increase in the expression of PTP1B protein in the spinal dorsal horn was inhibited by the treatment of i.t. injection of PTP1B siRNA. **** p < 0.0001 versus sham group; #### p < 0.0001 versus SNI + scRNA group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test. (C) Behavioral data showed that the established mechanical allodynia was reversed by the treatment of repeat i.t. injection of PTP1B siRNA, which started on day 7 after SNI. **** p < 0.0001 versus sham group; #### p < 0.0001 versus SNI + Vehicle or SNI + scRNA group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests. (D) A single dose of PTP1B‐IN‐1 (0.5 and 1 μg, i.t.) markedly increased the PWT in SNI rats (* p < 0.05, ** p < 0.01, **** p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests). (E) Repetitive injections of PTP1B‐IN‐1 (1 μg, i.t.) considerably reversed established mechanical allodynia in SNI rats (** p < 0.01, **** p < 0.0001 compared with Sham + Vehicle group; ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests). (F) The PWT was significantly increased from day 3 to day 8 in PTP1B‐IN‐1‐treated SNI rats compared with vehicle‐treated SNI rats (** p < 0.01, **** p < 0.0001 compared with Sham + Vehicle group; ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests).

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques: In Vitro, Expressing, Cell Culture, Control, Western Blot, Injection

Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: PTP1B‐IN‐1 inhibited the activation of ER stress and NF‐κB in rats with neuropathic pain. (A) PTP1B‐IN‐1 could decrease the elevated protein expression level of PTP1B in neuropathic pain rats (** p < 0.01 compared with Sham + Vehicle group, ## p < 0.01 compared with SNI + Vehicle group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test). (B–F) Western blot results indicated that PTP1B‐IN‐1 downregulated the elevated protein expression level of ER stress markers (BIP, p‐PERK/p‐eIF2α, p‐IRE1α, and ATF6) induced by neuropathic pain in the spinal cord (**** p < 0.0001 compared with Sham + Vehicle group; ## p < 0.01, ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test). (G) The subcellular morphological change of the neurons in the spinal dorsal horn after SNI. The yellow rows of the right panel indicate that the highly dilated ER membranes in the vehicle‐treated SNI rats were alleviated by PTP1B‐IN‐1 administration. Scale bars = 1 μm. (H) Western blot results indicated that PTP1B‐IN‐1 downregulated the elevated protein expression level of p‐NF‐κB induced by neuropathic pain in the spinal cord (**** p < 0.0001 compared with Sham + Vehicle group; #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test).

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques: Activation Assay, Expressing, Western Blot

Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: PTP1B‐IN‐1 inhibited the glial cell activation and the production of pro‐inflammatory cytokines in rats with neuropathic pain. (A, B) Western blot results showed that PTP1B‐IN‐1 ameliorated the upregulated protein expression level of Iba‐1 and GFAP in the spinal cord caused by neuropathic pain (**** p < 0.0001 compared with Sham + Vehicle group; #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test). (C–E) Western blot results indicated that treatment with PTP1B‐IN‐1 attenuated the increased protein expression level of TNF‐α, IL‐6, and IL‐1β in the spinal cord induced by neuropathic pain (*** p < 0.001, **** p < 0.0001 compared with Sham + Vehicle group; # p < 0.05, #### p < 0.0001 compared with SNI + Vehicle group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test).

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques: Activation Assay, Western Blot, Expressing

Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: Tm abolished PTP1B‐IN‐1‐mediated pain relief in SNI rats. (A) PTP1B‐IN‐1 alleviated the decreased PWT induced by neuropathic pain, while application with Tm could abolish the analgesic effect of PTP1B‐IN‐1 (**** p < 0.0001 compared with sham group; #### p < 0.001 compared with SNI + Vehicle group; &&&& p < 0.0001 compared with SNI + PTP1B‐IN‐1 group, n = 6 in each group, two‐way ANOVA, followed by Bonferroni tests). (B–F) PTP1B‐IN‐1 reduced the increased ER stress markers (BIP, p‐PERK/p‐eIF2α, p‐IRE1α, and ATF6) in neuropathic pain rats, while Tm reversed the downregulated BIP, p‐PERK/p‐eIF2α, p‐IRE1α, and ATF6 caused by PTP1B‐IN‐1 in neuropathic pain rats (* p < 0.05, *** p < 0.001, **** p < 0.0001 compared with sham group; # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group; && p < 0.01, &&& p < 0.001, &&&& p < 0.0001 compared with SNI + PTP1B‐IN‐1 group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test).

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques:

Journal: CNS Neuroscience & Therapeutics

Article Title: Protein tyrosine phosphatase 1B contributes to neuropathic pain by aggravating NF‐κB and glial cells activation‐mediated neuroinflammation via promoting endoplasmic reticulum stress

doi: 10.1111/cns.14609

Figure Lengend Snippet: Tm reversed the effect of PTP1B‐IN‐1 on NF‐κB, microglia, astrocytes activation, and neuroinflammation in the spinal cord of SNI rats. (A) PTP1B‐IN‐1 reduced the increased p‐NF‐κB, while Tm reversed the downregulated p‐NF‐κB caused by PTP1B‐IN‐1 in neuropathic pain rats (**** p < 0.0001 compared with sham group; #### p < 0.0001 compared with SNI + Vehicle group; &&&& p < 0.0001 compared with SNI + PTP1B‐IN‐1 group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test). (B–D) Western blot and immunofluorescence results showed that PTP1B‐IN‐1 ameliorated the upregulated protein expression levels of Iba‐1 and GFAP in the spinal cord and suppressed the activated microglia and astrocytes caused by neuropathic pain, while treatment with Tm abolished the effect of PTP1B‐IN‐1 on microglia and astrocyte activation (*** p < 0.001, **** p < 0.0001 compared with sham group; ### p < 0.001, #### p < 0.0001 compared with SNI + Vehicle group; &&& p < 0.001, &&& p < 0.001 compared with SNI + PTP1B‐IN‐1 group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test). (E–G) PTP1B‐IN‐1 reduced the increased TNF‐α, IL‐6, and IL‐1β, while Tm reversed the downregulated TNF‐α, IL‐6, and IL‐1β caused by PTP1B‐IN‐1 in neuropathic pain rats (** p < 0.01, *** p < 0.001 compared with sham group; # p < 0.05, ## p < 0.01, ### p < 0.001 compared with SNI + Vehicle group; && p < 0.01 compared with SNI + PTP1B‐IN‐1 group, n = 6 in each group, one‐way ANOVA followed by Bonferroni post‐hoc test).

Article Snippet: PTP1B‐IN‐1 (a potent inhibitor of PTP1B) and tunicamycin (Tm, an ER stress activator) were purchased from Medchemexpress (NJ, USA).

Techniques: Activation Assay, Western Blot, Immunofluorescence, Expressing