脊髓背角IL-17在神经病理性痛大鼠星形胶质细胞活化中的作用  

Role of interleukin-17 in spinal dorsal horns in neuropathic pain in rats and its effect on activation of astrocytes

在线阅读下载全文

作  者:孙彩霞[1] 袁雯[1] 李春叶[1] 刘嫣方[2] 陈盼[1] 苏兆亮[3] 许化溪[3] 

机构地区:[1]江苏大学附属医院麻醉科,镇江市212001 [2]镇江市第一人民医院中心实验室 [3]江苏大学免疫学研究所

出  处:《中华麻醉学杂志》2015年第3期320-325,共6页Chinese Journal of Anesthesiology

基  金:基金项目:国家自然科学基金(31270947);镇江市社会发展项目(SH2011028);江苏大学博士启动基金(jdfyRC-2013024)

摘  要:目的探讨脊髓背角IL—17在神经病理性痛大鼠星形胶质细胞活化中的作用。方法在体实验成年健康SPF级雄性SD大鼠64只,6—8周龄,体重180—200g,由江苏大学实验动物中心提供。采用随机数字表法,将其分为3组:对照组(C组,n=16)、假手术组(s组,n=24)和神经病理性痛组(NP组,n=24)。采用切断L5脊神经的方法制备神经病理性痛模型。于模型制备前、模型制备后1、3、5、7、10、时测定机械痛,阈61314d。于模型制备后7和14d,采用qRT—PCR法测定脊髓背角IL-17、IL-6、IL-1β、TNF-α的mRNA表达水平。于模型制备7d,测定脊髓背角星形胶质细胞的活化水平。离体实验采用随机数字表法将原代培养的乳鼠星形胶质细胞分为4组:空白对照组(C组,H=22)、10ng/ml IL-17组(110组,n=18)、50ng/ml IL-17组(150组,n=18)和100ng/ml IL-17组(I10组,n=22)。I10组、I50组和I10组分别用含上述3种浓度IL-17的培养基进行孵育,于孵育或培养24、48、72h时,采用MTT法检测星形胶质细胞的增殖水平。采用qRT—PCR法检测IL-6、IL-1β、TNF-α的mRNA表达水平。结果在体实验与C组比较,NP组模型制备后3—14d机械痛阈降低,模型制备后7d脊髓背角IL-17、IL-6、IL-1B的mRNA表达上调,星形胶质细胞活化水平升高(P〈0.05或0.01),s组模型制备后各时点机械痛阈差异无统计意义(P〉0.05)。离体实验与C组比较,I10组和I50组星形胶质细胞孵育48h时增殖水平升高,I100组孵育48和72h时星形胶质细胞增殖水平升高,IL-6、IL-1β的mRNA表达上调(P〈0.05或0.01),S组各时点星形胶质细胞增殖水平差异无统计学意义(P〉0.05)。结论脊髓背角IL-17表达上调可能参与了大鼠神经病理性痛的维持,其机制与促进星形胶质细胞活化,诱发中枢炎性反应有关。Objective To investigate the role of interleukin-17 (IL-17) in spinal dorsal horns in neuropathic pain (NP) in rats and its effect on activation of astrocytes. Methods In vivo experiment Sixty-four male SPF Sprague-Dawley rats, aged 6-8 weeks, weighing 180-200 g, were randomly divided into 3 groups using a random number table: control group (group C, n = 16), sham operation group (group S, n = 24) and group NP (n = 24). The animals were anesthetized with intraperitoneal pentobarbital sodium, the Ls.6 spinal nerves of the left side of the rat were gently separated and exposed, tightly ligated with 5-0 silk suture and transected. In group S, the Ls,6 spinal nerves of the left side of the rat were only exposed. In group C, no operation was performed. Mechanical pain threshold was measured at day 1 before operation and days 1, 3, 5, 7, 10 and 14 after operation. The expression of IL-17, IL-6, IL-Iβ and tumor necrosis factor-alpha (TNF-ot) mRNA in the spinal dorsal horn was determined using quantitative real-time PCR at day 7 and day 14 after operation. At day 7 after operation, the activation of astrocytes in the spinal dorsal horn was detected. In vitro experiment Primarily cultured astroeytes of neonatal rats were randomly divided into 4 groups using a random number table: control group ( group C, n=22), 10 ng/ml IL-17 group (Ilogroup, n=18), 50ng/ml IL-17 group (Isogroup, n = 18) and 100 ng/ml IL-17 group (I100 group, n= 22). In It0, Is0 and I10o groups, the astrocytes were incubated with the culture medium containing 10, 50 and 100 ng/ml IL-17, respectively. The proliferation of astrocytes was detected by MTT at 24, 48 and 72 h of ineutation or culture. The expression of IL-6, IL-1β and TNF-α mRNA was determined using quantitative real-time PCR. Results In vivo experiment Compared with group C, the mechanical pain threshold was significantly decreased at 3- 14 days after operation, the expression of IL-17, IL-6 and IL-1β mRNA in the spinal dorsal horn was up-regualt

关 键 词:白细胞介素17 神经痛 星形细胞 脊髓 

分 类 号:R614[医药卫生—麻醉学]

 

参考文献:

正在载入数据...

 

二级参考文献:

正在载入数据...

 

耦合文献:

正在载入数据...

 

引证文献:

正在载入数据...

 

二级引证文献:

正在载入数据...

 

同被引文献:

正在载入数据...

 

相关期刊文献:

正在载入数据...

相关的主题
相关的作者对象
相关的机构对象