Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: Expression of Iba1 in spinal cord of rats. ( A ) Thermal pain threshold of rats was assessed using the percentage of maximal possible antinociceptive effect (%MPE) according to the tail-flick latency. The %MPE in rats receiving morphine (10 μg, twice daily, intrathecally) on days 5 and 7 were decreased compared with the baseline on day 1. *** P <0.001, **** P <0.0001, vs NS rats. ( B, C ) The expressions of Iba1 mRNA ( B ) and protein ( C ) were significantly increased in morphine-tolerant rats measured by real-time PCR and Western blots, respectively. ** P <0.01, vs NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; MT, morphine tolerance; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Expressing, Tail Flick Test, Real-time Polymerase Chain Reaction, Western Blot, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: The effect of PDGFRβ agonist PDGF-BB on Iba1 expression. ( A ) PDGF-BB had no influence on %MPE of rats. ( B – D ) The phosphorylation of PDGFRβ ( B ) and the expressions of Iba1 mRNA ( C ) and protein ( D ) were significantly increased in rats received PDGF-BB measured by real-time PCR and Western blots, respectively. * P <0.05, ** P <0.01 vs PBS rats. ( E ) Rats were intrathecally injected with PDGF-BB or PBS once daily for 4 days, followed by a single dose of morphine on day 5. The %MPE in rats receiving PDGF-BB and morphine was decreased. ** P <0.01, vs PBS+Morphine rats. ( F ) The %MPE in rats receiving imatinib 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. ** P <0.01, vs Morphine+NS rats. ( G ) Pretreatment with imatinib inhibited the increased expression of Iba1 induced by morphine measured by Western blots. * P <0.05, vs NS+NS rats; # P <0.05, vs Morphine+NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; PBS, phosphate buffered saline; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Expressing, Phospho-proteomics, Real-time Polymerase Chain Reaction, Western Blot, Injection, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: The effect of MOR antagonist naloxone on activations of PDGFRβ and microglia. ( A ) The %MPE in rats receiving naloxone 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. * P <0.05, *** P <0.001, vs Morphine+DMSO rats. ( B, C ) Pretreatment with naloxone inhibited the phosphorylation of PDGFRβ ( B ) and the increased expression of Iba1 ( C ) induced by morphine measured by Western blots. * P <0.05, ** P <0.01, vs NS+DMSO rats; # P <0.05, vs Morphine+DMSO rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; DMSO, dimethyl sulfoxide; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Phospho-proteomics, Expressing, Western Blot, Saline

Journal: Journal of Pain Research

Article Title: The Interaction Between Spinal PDGFRβ and μ Opioid Receptor in the Activation of Microglia in Morphine-Tolerant Rats

doi: 10.2147/JPR.S255221

Figure Lengend Snippet: Involvement of JNK signaling in MOR-induced PDGFRβ activation in morphine tolerance. ( A ) Pretreatment with naloxone 30 minutes before morphine administration inhibited the phosphorylation of JNK induced by morphine measured by Western blots. ** P <0.01, vs NS+DMSO rats; # P <0.05, vs Morphine+DMSO rats. ( B ) The %MPE in rats receiving JNK inhibitor SP600125 30 minutes before morphine administration were higher than those in morphine-tolerant rats from day 5 to 7. ** P <0.01, vs Morphine+DMSO rats. ( C–E ) Pretreatment with SP600125 reduced increased expressions of p-c-Jun ( C ), p-PDGFRβ ( D ), and Iba1 ( E ) induced by morphine measured by Western blots. * P <0.05, *** P <0.001, vs NS+DMSO rats; # P <0.05, ## P <0.01, vs Morphine+DMSO rats. ( F ) Pretreatment with imatinib had no influence on the increased expression of p-JNK induced by morphine measured by Western blots. ** P <0.01, vs NS+NS rats. Values represent mean±SEM. n=6 in each group. Abbreviations: NS, normal saline; DMSO, dimethyl sulfoxide; %MPE, the percentage of maximal possible antinociceptive effect.

Article Snippet: The membranes were blocked with 5% bovine serum albumin for 2 hours at room temperature and incubated overnight at 4°C with mouse anti-ionized calcium-binding adapter molecule 1 (Iba1) antibody (1:200; Santa Cruz, USA), rabbit anti-p-JNK (Thr180/Tyr183) (1:1000; CST, USA), rabbit anti-phospho-PDGFRβ (1:1000; CST), rabbit anti-p-c-JUN (1:1000; CST), or rabbit anti-GAPDH (1:2000; Aspen, China).

Techniques: Activation Assay, Phospho-proteomics, Western Blot, Expressing, Saline