脊髓kindlin-1在大鼠神经病理性痛中的作用：与Wnt3a的关系

Role of spinal kindlin-1 in neuropathic pain in rats： the relationship with Wnt3a

目的评价脊髓kindlin．1在大鼠神经病理性痛中的作用及其与Wnt3a的关系。方法清洁级健康雄性sD大鼠18只，体重250～280g，10～12周龄，采用随机数字表法分为3组（n=6）：假手术组（S组）、神经病理性痛组（NP组）和kindlin．1抑制组（K组）。采用坐骨神经慢性压迫法建立大鼠神经病理性痛模型，s组仅暴露坐骨神经不结扎。K组于术前21d鞘内注射shRNA抑制kindlin．1表达，S组和NP组于术前21d鞘内注射空载病毒。分别于术前1d及术后1、4、7、10、13d时测定机械缩足反应阈（MWT）和热缩足潜伏期（TWL）。于术后13d时痛阈测定结束后处死大鼠取脊髓组织，采用Westernblot法检测脊髓kindlin，1和Wnt3a表达，RT-PCR法检测脊髓Wnt3amRNA表达。结果与S组比较，NP组术后4、7、10和13d时MWT降低，聊L缩短，脊髓Wnt3a及其mRNA、kindlin．1表达上调（P〈0．05）；与NP组比较，K组术后4、7、10和13d时MWT升高，TwL延长，脊髓Wnt3a及其mRNA、kindlin一1表达下调（P〈0．05）。结论kindlin．1通过上调Wnt3a表达参与大鼠神经病理性痛的发生发展过程。

Objective To evaluate the role of spinal kindlin-1 in neuropathic pain in rats and the relationship with Wnt3a. Methotls Eighteen clean-grade healthy male Sprague-Dawley rats, weighing 250-280 g, aged 10-12 weeks, were divided into 3 groups （n = 6 each） using a random number table： sham operation group （group S）, neuropathic pain group （group NP） and kindlin-1 inhibitor group （group K）. Neuropathic pain was induced by chronic compression of the sciatic nerve. The sciatic nerve was only exposed but not ligated in group S. In group K, shRNA was intrathecally injected at 21 days before opera- tion to inhibit the expression of kindlin-1. Vector virus was intratheeally injected at 21 days before operation in S and NP groups. The mechanical paw withdrawal threshold （MWT） and thermal paw withdrawal latency （TWL） were measured at 1 day before operation and 1, 4, 7, 10 and 13 days after operation. Rats were sacrificed at 13 days after measurement of pain threshold and the spinal cord was removed for determination of the expression of kindlin-1 and Wnt3a （ by Western blot） and expression of Wnt3a mRNA （ by real-time polymerase chain reaction）. Results Compared with group S, the MWT was significantly decreased and the TWL was shortened at 4, 7, 10 and 13 days, and the expression of Wnt3a protein and mRNA and kindlin-1 was up-regulated in group NP （P〈0. 05）. Compared with group NP, the MWT was significantly increased and the TWL was prolonged at 4, 7, 10 and 13 days, and the expression of Wnt3a protein and mRNA and kindlin-1 was down-regulated in group K （P〈0. 05）. Conclusion Kindlin-1 is involved in the development of neuropathic pain by up-regulating the expression of Wnt3a in rats.