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Feasibility of Signal Transducers and Activators of Transcription 3 Inhibitor WP1066 as a Novel Target for Neuropathic Pain
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摘要:
目的 观察Janus激酶2(Janus kinase 2, JAK2)/信号转导和转录激活子3(signal transducers and activators of transcription 3, STAT3)通路抑制剂WP1066对神经病理性疼痛的影响。 方法 12只体重180~200 g雌性SD大鼠用随机数字表法分为WP1066组及对照组(n=6), 均行双侧坐骨神经结扎手术, 两组于术前1 d、术前及术后第3、5天分别经鞘内注射容量为10 μl的WP1066(10 mmol/L溶于二甲基亚砜)或二甲基亚砜, 于术前及术后第3、5、7、10、14天进行动物行为学检测, 观察大鼠对机械、热及丙酮冷刺激的反应。术后第14天处死大鼠, 于L4~L6腰膨大脊髓背角处取材, 使用RT-PCR及Western blot检测STAT3、细胞因子信号抑制因子3(suppressor of cytokine signaling 3, SOCS3)、JAK2 mRNA及蛋白水平。 结果 大鼠机械缩足反射阈值、热刺激缩足反射潜伏期和冷痛阈值分别从第3、5、5天开始, WP1066组较对照组明显升高, 差异具有统计学意义(P < 0.05)。与对照组相比, WP1066组STAT3、SOCS3、JAK2 mRNA表达均明显降低(P < 0.05);WP1066组磷酸化STAT3、SOCS3、JAK2蛋白水平均亦明显降低(P < 0.05)。 结论 WP1066可明显改善大鼠神经病理性疼痛, 可能为治疗神经病理性疼痛提供新的靶点。 -
关键词:
- 神经病理性疼痛 /
- JAK/STAT通路 /
- WP1066 /
- 双侧坐骨神经结扎
Abstract:Objective To evaluate whether the Janus kinase 2(JAK2)/signal transducers and activators of transcription 3 (STAT3)inhibitor WP1066 could be a novel therapeutic target for neuropathic pain and its molecular mechnism. Methods Twelve female SD rats weighing 180-200 g were randomly divided into WP1066 group and control group(n=6) using the random number table. Rats in both groups underwent bilateral chronic sciatic nerve constriction injury (bCCI). WP1066 (10 μl, 10 mmol/L in dimethyl sulfoxide) or the equal volume of dimethyl sulfoxide was applied through the intrathecal tube one day before surgery, just before surgery, and 3 and 5 days after surgery in the WP1066 group and control group, respectively. Behavior tests were performed before surgery and 3, 5, 7, 10, and 14 days after the surgery to observe the rats' reactions to mechanical, thermal, and cold stimula-tions. The rats were killed on the 14th postoperative day. The dorsal horn of the spinal cord (L4-L6) was harvested, followed by the reverse transcription polymerase chain reaction (RT-PCR) and Western blotting to investigate the activation of the JAK2/STAT3 pathway. Results The pain-related behavior changes were significantly better in the WP1066 group than in the control group. WP1066 significantly inhibited the JAK2, suppressor of cytokine signaling 3(SOCS3), and STAT3 mRNA in rats with bCCI, and significantly decreased the ratio of JAK2, SOCS3 and phosphorylation of STAT3(p-STAT3) protein expression on the 14th postoperative day. Conclusions The administration of WP1066 can remarkably improve neuropathic pain in bCCI rats by inhibiting the STAT3 signaling pathway. Therefore, WP1066 may be a novel target for neuropathic pain. -
图 2 靶蛋白在大鼠脊髓背角的表达情况
A.pSTAT3；B.STAT3；C.SOCS3；D.JAK2
STAT3：信号转导和转录激活子3；pSTAT3：磷酸化STAT3；SOCS3：细胞因子信号抑制因子3；JAK2：Janus激酶2;Mr:相对分子质量；与对照组比较，*P < 0.05表 1 RT-PCR目的基因引物序列
引物名称 引物序列 STAT3-F 5′-TACCACAAAAGTCAGGTTGCTG-3′ STAT3-R 5′-ACATCCCCAGAGTCCTTATCAA-3′ SOCS3-F 5′-GAGAGCGGATTCTACTGGAGTG-3′ SOCS3-R 5′-GTCACTGCGCTCCAGTAGAAG-3′ JAK2-F 5′-GGTTCATTCAGCAGTTCAGTCA-3′ JAK2-R 5′-GCAGGGTCTCCAGGTTTATG-3′ IL-6-R 5′-TCTTGGGACTGATGTTGTTGAC-3′ IL-6-F 5′-TGGGTGGTATCCTCTGTGAAGT-3′ 
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