摘要
目的：观察尼氟灭酸（niflumic acid,NFA）干预后,TMEM16A在坐骨神经压榨性损伤（chronic constriction injury,CCI）模型鼠背根神经节（dorsal root ganglion,DRG）神经元上的表达,研究TMEM16A是否参与神经病理性疼痛。方法：制备CCI模型,热板实验检测热缩足反射潜伏期（thermal withdrawal latency,TWL）。在模型制备后第1 d开始以腹腔注射形式给予不同浓度的NFA干预14 d,取大鼠L4-6 DRG神经元进行实验。免疫荧光实验明确TMEM16A在DRG神经元中的分布;运用RTPCR技术和Western Blot技术在给予不同浓度NFA干预后,检测CCI模型DRG神经元上TMEM16A的mRNA水平和蛋白的表达。结果：（1）CCI组的TWL较正常组明显缩短,给予10μM、50μM和300μM的NFA干预后,各组TWL较CCI组均明显延长（P〈0.01,n=6）;50μM的NFA干预组较10μM的NFA干预组TWL也明显延长（P〈0.05,n=6）;但300μM的NFA干预组与50μM的NFA干预组相比,TWL时间差异无统计学意义。（2）免疫荧光显示TMEM16A主要存在于DRG神经元上;（3）CCI组mRNA水平较正常组明显增高;随着NFA浓度的增加,DRG神经元TMEM16A的mRNA水平较CCI组逐渐降低,差异具有统计学意义（P〈0.01,n=6）。（4）CCI组TMEM16A蛋白表达较正常组的明显增高;随着NFA浓度的增加,DRG神经元上TMEM16A蛋白表达较CCI组逐渐减低,差异具有统计学意义（P〈0.01,n=6）。结论：尼氟灭酸能抑制神经病理性疼痛大鼠DRG神经元TMEM16A的表达,而且与尼氟灭酸的剂量相关,提示钙激活氯通道可能参与或调控神经病理性疼痛的发生。
Objective： To observe expression of TMEM16 A in DRG neurons in rats with Chronic Constriction Injury of the sciatic nerve（CCI） after treatment of niflumic acid（NFA）, and explore whether TMEM16 A is involved in the neuropathic pain. Methods： After CCI surgery, we measured thermal withdrawal latency（TWL） using hot plate test. Intraperitoneal injection of NFA with different concentrations at 14 day after CCI operation, took the L4-6 DRG out of rat for the following research.Immunofluorescence assay was used to show the distribution of TMEM16 A in the DRG neurons; RT-PCR and Western Blot techniques were used to detect the expression of mRNA and protein of TMEM16 A on DRG neurons in different groups. Results：（1） TWL of CCI group was significantly shorter than normal group, the TWL of each NFA groups were longer than the CCI group（P〈0.01, n = 6）; the TWL of 50 μM NFA group was significantly longer than the 10μM NFA group（P〈0.05, n = 6）; but there were no statistical differences between the 300 μM and 50 μM NFA groups.（2） Immunofluorescence showed TMEM16 A were mainly in the DRG neurons;（3） mRNA level of CCI group was significantly higher than that of normal group. With the increased concentrations of NFA mRNA levels of TMEM16 A on DRG neurons decreased gradually compared with CCI group, the level had statistically significant（P〈0.01, n = 6）.（4） The expression of TMEM16 A protein on DRG in CCI group was significantly higher than the normal group; With the increased concentrations of NFA, expression of TMEM16 A protein on DRG neurons gradually reduced compared with CCI group, the difference has statistically significance（P〈0.01, n = 6）. Conclusion： NFA can inhibit the expression of TMEM16 A on DRG neurons of neuropathic pain rats, which was associated with the amount of NFA,which prompt that calcium-activated chloride channel may be involved in the occurrence or the regulation of neuropathic pain.
出处
《中国疼痛医学杂志》
CAS
CSCD
2015年第11期820-825,共6页
Chinese Journal of Pain Medicine
基金
兵团优秀青年创新基金专项(2010JC33)
GABA介导的初级感觉传入突触前抑制在神经病理性疼痛中去抑制的机制研究
