title: STAT3 inhibitor WP1066 as a novel therapeutic agent for bCCI neuropathic pain rats.
abstract: Activation of signal transducer and activator of transcription-3 (STAT3) is suggested to be critically involved in the development of chronic pain, but the complex regulation of STAT3-dependent pathway and the functional significance of inhibiting this pathway during the development of neuropathic pain remain elusive. To evaluate the contribution of the JAK2/STAT3 pathway to neuropathic pain and the potentiality of this pathway as a novel therapeutic target, we examined the effects of the STAT3 inhibitor WP1066 by intrathecal administration in a rat model of bilateral chronic constriction injury (bCCI). The pain behavior tests were performed before the surgery and on postoperative day 3, 7, 14 and 21. L4-L6 dorsal spinal cord were harvested at each time point. Both RT-PCR and Western blot were performed to evaluate the activation of JAK2/STAT3 pathway. To observe the influence of WP1066 on neuropathic pain and its molecular mechanism, WP1066 (10 μl, 10 mmol/L in DMSO) or the same capacity of DMSO as the control were applied through the intrathecal tube on the day before bCCI surgery, and on the postoperative day 3 and 5. Behavioral tests were performed to observe the therapeutic effect on mechanical, thermal and cold hyperalgesia. L4-L6 dorsal spinal cord was harvested on postoperative day fourteen, followed by RT-PCR and Western blot evaluation of the JAK2/STAT3 pathway activation. The mechanical, thermal and cold hyperalgesia of the bCCI rats were significantly decreased when compared with the Sham or the Naïve group at each postoperative time point (P<0.05). JAK2 mRNA and STAT3 mRNA were significantly increased in the bCCI rats, accompanied by SOCS3 mRNA with a similar tendency. Western blot analysis showed that JAK2 and phosphorylated STAT3 increased significantly since 3 days after bCCI. JAK2 peaked on postoperative day 14 while phosphorylated STAT3 peaked on postoperative day 7 and gradually decreased thereafter and SOCS3׳s peak level on postoperative day 3. When WP1066 were administered intrathecally, the pain behaviors of the bCCI rats were significantly improved (P<0.05). WP1066 also inhibited the mRNA level of JAK2, STAT3 and SOCS3 in bCCI rats significantly, together with the protein level of JAK2, phosphorylated STAT3 and SOCS3 on postoperative day 14 as well. Our results found that the JAK2/STAT3 pathway in the spinal cord dorsal horn was significantly activated in the bCCI neuropathic pain rats. WP1066, which inhibited the STAT3 pathway specifically, could partially alleviate the pain behavior of the bCCI rats. So it may serve as a novel therapeutic strategy against neuropathic pain.
authors.affiliation: Department of Anesthesiology, Peking Union Medical College Hospital, CAMS&PUMC, Beijing 100730, China. Electronic address: zhaojingxuepumch@163.com.
authors.forename: Zhao-Jing
authors.initials: ZJ
authors.lastname: Xue
authors.affiliation: Department of Anesthesiology, Peking Union Medical College Hospital, CAMS&PUMC, Beijing 100730, China. Electronic address: shenle@pumch.cn.
authors.forename: Le
authors.initials: L
authors.lastname: Shen
authors.affiliation: Department of Anesthesiology, Peking Union Medical College Hospital, CAMS&PUMC, Beijing 100730, China. Electronic address: wangzhiyao0313@163.com.
authors.forename: Zhi-Yao
authors.initials: ZY
authors.lastname: Wang
authors.affiliation: Department of Anesthesiology, Peking Union Medical College Hospital, CAMS&PUMC, Beijing 100730, China. Electronic address: huishangyi@126.com.
authors.forename: Shang-Yi
authors.initials: SY
authors.lastname: Hui
authors.affiliation: Department of Anesthesiology, Peking Union Medical College Hospital, CAMS&PUMC, Beijing 100730, China. Electronic address: garybeijing@163.com.
authors.forename: Yu-Guang
authors.initials: YG
authors.lastname: Huang
authors.affiliation: Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, Department of Anatomy, Histology and Embryology, Beijing 100005, China. Electronic address: mcpumc@163.com.
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
mesh_terms: D000700:Analgesics / Q000494:pharmacology*; D000818:Animals; D002908:Chronic Disease; D003080:Cold Temperature; D003251:Constriction, Pathologic; D004195:Disease Models, Animal; D005260:Female; D006358:Hot Temperature; D006930:Hyperalgesia / Q000188:drug therapy / Q000503:physiopathology; D053614:Janus Kinase 2 / Q000378:metabolism; D008159:Lumbar Vertebrae; D009437:Neuralgia / Q000188:drug therapy* / Q000503:physiopathology; D011725:Pyridines / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D011897:Random Allocation; D017207:Rats, Sprague-Dawley; D050796:STAT3 Transcription Factor / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015398:Signal Transduction; D013116:Spinal Cord / Q000187:drug effects / Q000503:physiopathology; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins / Q000378:metabolism; D014110:Touch; D020032:Tyrphostins / Q000494:pharmacology*
chemical_list: D000700:Analgesics; D011725:Pyridines; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor; C491411:Socs3 protein, mouse; C494087:Stat3 protein, mouse; D000071223:Suppressor of Cytokine Signaling 3 Protein; D050826:Suppressor of Cytokine Signaling Proteins; D020032:Tyrphostins; C519885:WP1066; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2
keywords: JAK2; Neuropathic pain; STAT3; WP1066