Journal: Molecules

Article Title: Syringaresinol Alleviates Oxaliplatin-Induced Neuropathic Pain Symptoms by Inhibiting the Inflammatory Responses of Spinal Microglia

doi: 10.3390/molecules27238138

Figure Lengend Snippet: Oxaliplatin treatment activated microglia in BV-2 resulting in the increased expression of inflammatory mediators that can induce pain behavior when intrathecally injected, which was suppressed by syringaresinol treatment. ( A ) The chemical structure of syringaresinol. ( B – I ) Western blot analysis of representative proinflammatory cytokines (IL-1β, IL-6, and TNF-α), enzymes (COX-2 and iNOS), and signaling molecules (p-ERK and p-NF-kB) in the BV-2 cell line. Ratio of protein expressions relative to GAPDH or total forms (ERK and NF-kB) expression. Syringaresinol treatment significantly suppressed the oxaliplatin-induced upregulation of inflammatory mediators. # p < 0.05 and ## p < 0.01 vs. vehicle treated BV-2 cells; * p < 0.05 and ** p <0.01 vs. Oxa treated BV-2 cells with one-way ANOVA with a Tukey post hoc test to compare multiple groups. ( J – M ) A single intrathecal injection of Oxa BV-2 serum elicited cold ( J ) and mechanical allodynia ( K ) in naïve mice. Vehicle BV-2 Serum: n = 5, Oxa BV-2 Serum: n = 9. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. Vehicle BV-2 Serum with two-way ANOVA followed by Sidak’s post-test for multiple comparisons. ( N ) Intrathecal injection of syringaresinol completely canceled out cold allodynia induced by Oxa BV-2 serum. ( O ) Mechanical allodynia induced by Oxa BV-2 serum was significantly ameliorated by syringaresinol treatment. Oxa BV-2 Serum + vehicle: n = 6, Oxa BV-2 Serum + syringaresinol: n = 6. *** p < 0.001, **** p < 0.0001 vs. Baseline and # p < 0.05, #### p < 0.0001 vs. Oxa BV-2 Serum + vehicle with two-way ANOVA followed by Sidak’s post-test for multiple comparisons.

Article Snippet: Samples were then incubated with primary antibodies [rabbit anti-interleukin-1 beta (IL-1ß), mouse anti-IL-6, rabbit anti-tumor necrosis factor-α (TNF-α) (Cell Signaling Technology, Danvers, MA, USA), mouse anti-cyclooxygenase-2 (COX-2), mouse anti-iNOS (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit anti-phospho-extracellular signal-regulated kinase 1/2 (p-ERK), rabbit anti-ERK, rabbit anti-phospho-nuclear factor kappa B (p-NF-κB), rabbit anti-NF-κB, and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell Signaling Technology)] diluted in 3% BSA (1: 1000) overnight at 4 °C.

Techniques: Expressing, Injection, Western Blot

Journal: Molecules

Article Title: Syringaresinol Alleviates Oxaliplatin-Induced Neuropathic Pain Symptoms by Inhibiting the Inflammatory Responses of Spinal Microglia

doi: 10.3390/molecules27238138

Figure Lengend Snippet: A single injection of syringaresinol downregulates co-localization of Iba 1 and iNOS, p-ERK, or p-NF-kB at the spinal dorsal horn. ( A – G ) Representative images of the spinal dorsal horn and a summarized graph showing the colocalizaion of Iba 1 and iNOS; ( A , D ) iNOS, ( B , E ) Iba 1, and ( C , F ) merged image. ( G ) Comparison between co-localization. Syringaresinol treatment significantly reduced the co-localization of Iba 1 and iNOS in oxaliplatin-injected animals. ( H – N ) Representative images of -the spinal dorsal horn and a summarized graph showing the co-localization of Iba 1 and p-ERK. ( H , K ) p-ERK, ( I , L ) Iba 1, and ( J , M ) merged image. ( N ) Syringaresinol treatment significantly reduced the co-localization of Iba 1 and p-ERK. ( O – Q ) Representative image of the spinal dorsal horn and a summarized graph showing the co-localization of Iba 1 and p-NF-κB. ( O , R ) p-NF-κB, ( P , S ) Iba 1, and ( Q , T ) merged image. ( U ) Syringaresinol treatment significantly reduced the co-localization of Iba 1 and p-NF-κB. For iNOS, Oxa: n = 4, and Oxa + Syr: n = 5. For, p-ERK, Oxa: n = 6, Oxa + Syr: n = 6. For p-NF-κB, Oxa: n = 6, Oxa + Syr: n = 6. * p < 0.05, ** p < 0.01 by unpaired t-test.

Article Snippet: Samples were then incubated with primary antibodies [rabbit anti-interleukin-1 beta (IL-1ß), mouse anti-IL-6, rabbit anti-tumor necrosis factor-α (TNF-α) (Cell Signaling Technology, Danvers, MA, USA), mouse anti-cyclooxygenase-2 (COX-2), mouse anti-iNOS (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit anti-phospho-extracellular signal-regulated kinase 1/2 (p-ERK), rabbit anti-ERK, rabbit anti-phospho-nuclear factor kappa B (p-NF-κB), rabbit anti-NF-κB, and rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Cell Signaling Technology)] diluted in 3% BSA (1: 1000) overnight at 4 °C.

Techniques: Injection