白细胞介素-10通过乙二醛酶1影响脊髓钝挫伤大鼠运动功能的恢复

doi:10.13418/j.issn.1001-165x.2021.03.014

中国临床解剖学杂志 ›› 2021, Vol. 39 ›› Issue (3): 323-329.doi: 10.13418/j.issn.1001-165x.2021.03.014

白细胞介素-10通过乙二醛酶1影响脊髓钝挫伤大鼠运动功能的恢复

牛英杰¹，　董越娇¹，　王若遥¹，　赵鑫鑫¹，　母姝祺¹，　张晓²，　荣成²

成都医学院　1.临床医学院；2.基础医学院实验教学中心，成都 610500

收稿日期:2019-10-25 出版日期:2021-05-25 发布日期:2021-06-02
通讯作者: 张晓，教授，硕士研究生导师，E-mail：954073462@qq.com；荣成，研究员，E-mail：344964514@qq.com
作者简介:牛英杰（1995-），在读研究生，研究方向：神经系统损伤与修复，E-mail： 18328081791@163.com
基金资助:
四川省大学生创新创业训练项目（201813705061）

Interleukin-10 affects the recovery of motor function of spinal cord contusion rats via glyoxalase-1

Niu Yingjie¹, Dong Yuejiao¹, Wang Ruoyao¹, Zhao Xinxin¹, Mu Shuqi¹, Zhang Xiao¹, Rong Cheng¹

1. Clinical Medicine College, 2. Experiment Technology Center, Preclinical Medicine of Chengdu Medical College, Chengdu 610500, Sichuan Province, China

Received:2019-10-25 Online:2021-05-25 Published:2021-06-02

摘要/Abstract

摘要： 目的　用慢病毒介导的方法抑制白细胞介素-10（IL-10）的RNA，观察IL-10通过乙二醛酶1（GLO1）影响大鼠脊髓损伤后运动功能恢复。方法　SD大鼠66只，采用Allen’s法制备脊髓钝挫伤（spinal cord contusion，SCC）模型，随机分为Sham组（n=9）、SCC组（n=27）、Vector组（n=15）和IL-10 SH组（n=15），分别注射空质粒和包装IL-10 siRNA的质粒；每组再分为3 d、7 d和28 d 亚组。采用BBB（Basso，Beattie，Bresnahan）评分评估大鼠后肢运动功能；免疫组织化学和qRT-PCR技术定位和定量测定IL-10和GLO1的表达变化；用慢病毒包装的RNA构建IL-10的RNA干扰模型（IL-10-SH-LV）评估IL-10和GLO1在脊髓损伤后的功能，GeneMANIA生物信息学预测IL-10和GLO1的关系。结果　SCC后大鼠双后肢截瘫，后肢运动功能消失，BBB评分在损伤后1 d，3 d，7 d，14 d逐渐恢复，但低于对照组。qRT-PCR结果显示IL-10在脊髓损伤后表达显著降低；SCC后，免疫组织化学结果显示IL-10主要在脊髓前角的神经元和神经胶质细胞中表达。慢病毒干扰IL-10的表达后，随着时间推移，GLO1表达增加，大鼠运动功能BBB评分也逐渐恢复。GeneMANIA结果显示GLO1和IL-10通过共表达的Hpd和Klk1c2、Kcns1、Proc相互作用。结论　IL-10通过上调抗氧化应激因子GLO1的表达促进脊髓损伤大鼠运动功能恢复。

关键词: 白细胞介素-10, 　乙二醛酶1, 　RNAi, 　大鼠, 　脊髓钝挫伤

Abstract: Objective　To inhibit the expression of IL-10 and to observe IL-10 affected the recovery of motor function of spinal cord contusion rats by glyoxalase-1 by using lentivirus mediated RNA.　Methods　The model of spinal cord contusion (SCC) was established in 66 Sprague-Dawley rats with Allen's method. The rats were randomized into a Sham group (n=9); a SCC group (n=27)、a Vector group (n=15) and a IL-10 SH group (n=15), which were injected blank lentivirus vector and vector of IL-10 siRNA respectively and then divided into 3 days, 7 days and 28 days subgroups. Basso, Beattie, Bresnahan (BBB) score was used to evaluate the motor function recovery of hind limbs in rats. The localization and expression of GLO1 were examined by immunohistochemistry and real-time quantitative polymerase chain reaction (qRT-PCR) respectively. The inhibition of IL-10 (IL-10-SH-LV) mediated by lentivirus transduction was used to elucidate the function of IL-10 and GLO1. GeneMANIA was used to predict the relationship between IL-10 and GLO1.　Results　The motor capability of hind limbs disappeared after SCC and followed by recovery at 1d, 3d, 7d and 14d as evaluated by BBB score, but it still could not reach the normal level. The expression of IL-10 decreased obviously in spinal cord tissue after SCC was detected by qRT-PCR, and the immunoreactive positive reactant of IL-10 was located in neurons and neuroglial cells of ventral horn in spinal cord by immunofluorescence. Inhibition of IL-10 mediated by lentivirus reduced motor function (evaluated by BBB score), but increased in the next time with the upregulation of GLO1. Futher more, GeneMANIA result showed that GLO1 contacted with IL-10 through Hpd and Klk1c2/Kcns1/Proc, which were co-expression in rats.　Conclusions　IL-10 promotes functional recovery of spinal cord contusion rats, which may be associated with upregulation of GLO1.

Key words: IL-10;　GLO1, 　RNAi;　Rats;　Spinal cord contusion

中图分类号:

R744

牛英杰, 董越娇, 王若遥, 赵鑫鑫, 母姝祺, 张晓, 荣成. 白细胞介素-10通过乙二醛酶1影响脊髓钝挫伤大鼠运动功能的恢复[J]. 中国临床解剖学杂志, 2021, 39(3): 323-329.

Niu Yingjie, Dong Yuejiao, Wang Ruoyao, Zhao Xinxin, Mu Shuqi, Zhang Xiao, Rong Cheng. Interleukin-10 affects the recovery of motor function of spinal cord contusion rats via glyoxalase-1[J]. Chinese Journal of Clinical Anatomy, 2021, 39(3): 323-329.

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白细胞介素-10通过乙二醛酶1影响脊髓钝挫伤大鼠运动功能的恢复

Interleukin-10 affects the recovery of motor function of spinal cord contusion rats via glyoxalase-1

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