鲁义++++++姚嘉茵++++++尧新华++++++卿朝晖++++++童辉4++++刘玲

[摘要] 目的 探讨Toll样受体4（TLR4）抑制剂TAK-242对糖尿病周围神经痛（DPN）模型大鼠的治疗效果及其可能的作用机制。 方法 SPF级雄性SD大鼠60只，随机均分为3组，分别为正常对照组（NC组）、DPN模型组（DPN组）、TAK-242治疗组（TAK组）。采用链脲佐菌素（STZ）方法建立DPN大鼠模型，采用ELISA及RT-PCR方法检测DPN模型大鼠腰膨大脊髓组织的高迁移率族蛋白B1（HMGB1）-TLR4轴上下游基因（HMGB1、TLR4、MAPK、NF-κB、IL-6）的变化，分析上述细胞因子表达水平与大鼠疼痛行为的相关性。使用TLR4抑制剂TAK-242对DPN模型大鼠进行药物干预，观察其治疗效果及对HMGB1-TLR4轴基因表达的影响。 结果 ELISA检测显示，DPN组大鼠的血清HMGB1、TLR4、IL-6表达水平均较NC组升高（P<0.05）；TAK组大鼠的TLR4及IL-6表达水平较DPN组下降（P<0.05）。RT-PCR检测显示，DPN组大鼠的HMGB1、TLR4、NF-κB、IL-6 mRNA表达水平较NC组升高（P<0.05）；TAK组大鼠的TLR4、NF-κB、IL-6 mRNA表达水平较DPN组下降（P<0.05）。 结论 TLR4抑制剂TAK-242通过阻断HMGB1-TLR4轴对DPN模型动物起治疗作用。

[关键词] TLR4；TAK-242；HMGB1-TLR4；糖尿病周围神经痛

[中图分类号] R587.2 [文献标识码] A [文章编号] 1674-4721（2016）02（a）-0009-04

Therapeutic effect of TAK-242 on diabetic peripheral neuropathic pain rats

LU Yi1 YAO Jia-yin2 YAO Xin-hua1▲ QING Zhao-hui1 TONG Hui1 LIU Ling1

1.Department of Anesthesiology，Guangzhou Hospital of Traditional Chinese Medicine，Guangzhou 510130，China；2.Department of Gastroenterology，the Sixth Hospital Affiliated to Sun Yat-Sen University，Guangzhou 510655，China

[Abstract] Objective To investigate the therapeutic effect and possible mechanisms of TLR4 inhibitor named TAK-242 on the diabetic peripheral neuropathic （DPN） rat model. Methods 60 male SD rats with SPF level were randomly divided into 3 groups， such as normal control group （NC group）， DPN model group （DPN group），TAK-242 treatment group （TAK group）.DPN rat models were established by using the method of streptozocin （STZ），and behavior analysis were made.Expressions of HMGB1-TLR4 related genes such as HMGB1，TLR4，MAPK，NF-κB and interleukin-6 （IL-6） were determined by RT-PCR.Correlations of genes mentioned above and the pain behavior of rats were analyzed.TLR4 inhibitor named TAK-242 was used for drug intervention.Its therapeutic effect and the effect on the expressions of HMGB1-TLR4 axis were observed. Results There was a significant increase in the spontaneous behavior and decrease in the pain threshold of rats in DNP group compared with NC group （P<0.05）.The serum expression of TLR4 and IL-6 in DNP group was significantly higher than those in NC group （P<0.05），and the expression of IL-6 and TLR4 in the TAK group was decreased compared with the model group as detected by ELISA （P<0.05）.The expressions of HMGB1，TLR4，NF-κB and IL-6 mRNA in DNP group was significantly higher than those in NC group （P<0.05），and the mRNA expression of TLR4，NF-κB and IL-6 in TAK group was decreased compared with DNP group as detected by RT-PCR （P<0.05）. Conclusion By blocking the HMGB1-TLR4 axis，TLR4 inhibitor TAK-242 plays a role of therapeutic effect in the treatment on DPN rats.

[Key words] TLR4；TAK-242；HMGB1-TLR4；Diabetic peripheral neuropathic

在糖尿病患者中约20%的患者合并有糖尿病周围神经病变，占糖尿病神经病变的50%[1-2]，其中糖尿病周围神经痛（diabetic peripheral neuropathy，DPN）发病率为13%～26%[3-4]。最新研究认为DPN的发生、发展与全身慢性炎性反应密切相关，尤其周围神经局部免疫功能失常、细胞因子信号传导轴异常相关[5]。其中，高迁移率族蛋白B1（high mobility group box-1 protein，HMGB1）-Toll样受体4（Toll-like receptor 4，TLR4）轴可能是神经免疫性损伤的核心病因[6]。本研究拟探讨HMGB1-TLR4轴在DPN发病机制中的作用，使用TLR4抑制剂TAK-242阻断HMGB1-TLR4轴，观察其对DPN的治疗效果。

1 材料与方法

1.1 实验动物

雄性SD大鼠60只，SPF级，体质量180～220 g，由中山大学疼痛治疗中心提供；安排专人采用专用饲料和水进行喂养。实验动物质量合格证：SCXK（粤）2009-0011。实验动物设施使用证：编号0044336。

1.2动物分组

60只大鼠随机均分为3组，分别设为正常对照组（normal control group，NC组）、DPN模型组（DPN model group，DPN组）、TAK-242治疗组（TAK-242 treatment group，TAK组）。

1.3动物模型的建立