摘要目的:评价大鼠神经病理性痛时脊髓kindlin-1/Wnt3a信号通路与炎症反应的关系。方法:清洁级健康雄性SD大鼠24只，10～12周龄，体重250～280 g，采用随机数字表法分为4组( n＝6)：假手术组(SH组)、神经病理性痛组(NP组)、kindlin-1 shRNA组(K组)和Wnt3a抑制组(W组)。采用慢性坐骨神经压迫法建立小鼠神经病理性痛模型。于术前21 d时K组鞘内注射kindlin-1 shRNA腺病毒载体10 μl，SH组、NP组和W组鞘内注射病毒空载体10 μl。于术后1～3 d时W组鞘内注射Wnt抑制剂IWP-2 10 μl，SH组、NP组和K组鞘内注射人工脑脊液10 μl。分别于术前1 d、术后4和7 d时测定机械缩足反应阈(MWT)和热缩足潜伏期(TWL)。术后7 d痛阈测定结束后，处死大鼠取L 4-6脊髓组织，采用ELISA法测定TNF-α和IL-1β含量，采用Western blot法检测kindlin-1和Wnt3a表达。 结果:与SH组比较，NP组、K和W组术后4和7 d时MWT降低，TWL缩短，脊髓TNF-α和IL-1β含量升高，NP组和W组脊髓kindlin-1和Wnt3a表达上调，K组脊髓Wnt3a表达上调( P<0.05)；与NP组比较，K组和W组术后4和7 d时MWT升高，TWL延长，K组脊髓TNF-α和IL-1β含量降低，kindlin-1和Wnt3a表达下调，W组脊髓TNF-α和IL-1β含量降低，Wnt3a表达下调( P<0.05)，kindlin-1表达差异无统计学意义( P>0.05)。 结论:脊髓kindlin-1通过上调Wnt3a的表达，进而调控炎症反应，参与大鼠神经病理性痛的维持。

abstractsObjective:To evaluate the relationship between spinal kindlin-1/Wnt3a signaling pathway and inflammatory response in a rat model of neuropathic pain (NP).Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 10-12 weeks, weighing 250-280 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (SH group), NP group, kindlin-1 shRNA group (K group) and Wnt3a inhibition group (W group). NP was induced by chronic constrictive injury in anesthetized animals.At 21 days before operation, kindlin-1 shRNA adenovirus vector 10 μl was intrathecally injected in group K, and empty viral vector 10 μl was intrathecally injected in SH, NP and W groups.Wnt inhibitor IWP-2 10 μl was intrathecally injected in group W, and artificial cerebrospinal fluid 10 μl was intrathecally injected in SH, NP and K groups at 1-3 days after operation.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before operation and 4 and 7 days after operation, respectively.At the end of pain threshold measurement at 7 days after operation, the animals were sacrificed and the lumbar segments (L 4-6) of the spinal cord were obtained for determination of the contents of tumor necrosis factor (TNF)-α and interleukin (IL)-1β (IL-1β) (using enzyme-linked immunosorbent assay) and the expression of kindlin-1 and Wnt3a (by Western blot). Results:Compared with group SH, MWT was significantly decreased, TWL was shortened at 4 and 7 days after operation, and the contents of TNF-α and IL-1β in spinal cord were increased in NP, K, and W groups, the expression of kindlin-1 and Wnt3a was up-regulated in NP and W groups, and expression of Wnt3a was up-regulated in group K ( P<0.05). Compared with group NP, MWT was significantly increased and TWL was prolonged at 4 and 7 days after operation in K and W groups, the contents of TNF-α and IL-1β in spinal cord were decreased, and the expression of kindlin-1 and Wnt3a was down-regulated in group K, the contents of TNF-α and IL-1β in spinal cord were decreased, and the expression of Wnt3a was down-regulated in group W ( P<0.05), and no significant change was found in kindlin-1 expression ( P>0.05). Conclusion:Spinal kindlin-1 regulates the inflammatory response by up-regulating the expression of Wnt3a, and it is involved in the maintenance of NP in rats.