富血小板血浆通过Nrf2信号通路对脊髓损伤小鼠的抗炎抗氧化作用及其机制  

Anti-inflammatory and antioxidant effects of platelet-rich plasma on mice with spinal cord injury through Nrf2 signaling pathway

作　　者：寇红伟[1] 闫立言 韩萧男 张明康 郑赛磊 陈松峰 尚国伟[1] 姬彦辉 冷子宽 程田[1] 刘宏建[1] Kou Hongwei;Yan Liyan;Han Xiaonan;Zhang Mingkang;Zheng Sailei;Chen Songfeng;Shang Guowei;Ji Yanhui;Leng Zikuan;Cheng Tian;Liu Hongjian(Department of Orthopaedics,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院骨科,450052

出　　处：《中华实验外科杂志》2022年第11期2081-2083,共3页Chinese Journal of Experimental Surgery

基　　金：河南省高等学校重点科研项目计划 (21A320047)。

摘　　要：目的探讨富血小板血浆通过Nrf2信号通路对脊髓损伤小鼠的抗炎抗氧化作用及其作用机制。方法采用改良Allen’s击打法制备小鼠脊髓损伤模型,采用随机数字表法将小鼠分为对照组(SHAM组)、脊髓损伤组(SCI组)、富血小板血浆处理组(PRP组)。术后7 d,小鼠麻醉处死,酶联免疫吸附试验(ELISA)法检测白细胞介素(IL)-1β、IL-10的表达,氧化应激试剂盒检测超氧化物歧化酶(SOD)、丙二醛(MDA)和还原型谷胱甘肽(GSH)的含量,蛋白质印迹法(Western blot)检测Nrf2蛋白和Keap1蛋白的表达,尼氏染色观察损伤组织中尼氏体的数量,并使用小鼠BMS评分于损伤后1、3、7、14、21、28 d评估运动功能。组间比较采用t检验和单因素方差分析。结果(1)PRP组尼氏体数量[(28.50±5.68)个]明显高于SCI组[(12.83±5.04)个],差异有统计学意义(t=5.053,P<0.05)。(2)PRP组SOD[(131.07±15.45)U/g]和GSH[(5.60±0.98)μg/g]含量高于SCI组[(102.51±24.00)U/g、(2.28±0.58)μg/g],差异有统计学意义(t=-2.451、-7.126,P<0.05)。MDA[(39.17±13.53)nmol/g]含量低于SCI组[(76.14±12.68)nmol/g],差异有统计学意义(t=4.882,P<0.05)。(3)PRP组促炎因子IL-1β表达量[(85.06±15.45)pg/ml]低于SCI组[(121.28±19.98)pg/ml],而抑炎因子IL-10表达量[(607.14±41.59)pg/ml]高于SCI组[(480.48±42.06)pg/ml],差异有统计学意义(t=3.513、-5.245,P<0.05)。(4)相较于SCI组(0.79±0.15、1.69±0.19),PRP组的Nrf2蛋白表达量增加(1.11±0.23),Keap1蛋白表达量降低(1.27±0.15),差异有统计学意义(t=-2.846、4.350,P<0.05)。(5)脊髓损伤后第14、21、28天,PRP组BMS评分[(2.50±0.55)、(3.17±0.75)、(3.83±0.75)分]高于SCI组[(1.50±0.84)、(2.17±0.75)、(2.50±0.55)分],差异有统计学意义(t=-2.449、-2.301、-3.508,P<0.05)。结论富血小板血浆通过Nrf2信号通路对脊髓损伤小鼠发挥抗炎抗氧化作用。Objective To investigate the anti-inflammatory and antioxidant effect of platelet-rich plasma on spinal cord injury mice through the Nrf2 signaling pathway and its related mechanism.Methods The mouse spinal cord injury model was established by modified Allen′s shock method.The mice were divided into SHAM group(SHAM group),SCI group(SCI group)and platelette-rich plasma treatment group(PRP group)by random number table method.The expression of interleukin(IL)-1β,IL-10 and were detected by enzyme linked immunosorbent assay(ELISA).The content of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)were detected by oxidative stress kit.The expression of the Nrf2 protein and Keap1 protein were detected by Western blotting.The number of nissenite in the injured tissues was observed by nissenite staining.The BMS motor function scores of mice were evaluated after injury at 1,3,7,14,21 and 28 days.T-test and one-way ANOVA were used for comparison between groups.Results(1)The number of Nissile bodies in PRP group(28.50±5.68)was higher than that in SCI group(12.83±5.04),and the difference was statistically significant(t=5.053,P<0.05).(2)SOD[(131.07±15.45)U/g]and GSH[(5.60±0.98)μg/g]in PRP group were higher than those in SCI group[(102.51±24.00)U/g,(2.28±0.58)μg/g].The difference was statistically significant(t=-2.451,-7.126,P<0.05).MDA[(39.17±13.53)nmol/g]was lower than that in SCI group[(76.14±12.68)nmol/g].The difference was statistically significant(t=4.882,P<0.05).(3)The expression of pro-inflammatory factor IL-1βin PRP group[(85.06±15.45)pg/ml]was lower than that in SCI group[(121.28±19.98)pg/ml],while the expression of anti-inflammatory factor IL-10 in PRP group[(607.14±41.59)pg/ml]was higher than that in SCI group[(480.48±42.06)pg/ml],and the difference was statistically significant(t=3.513,-5.245,P<0.05).(4)Compared with SCI group(0.79±0.15,1.69±0.19),the protein expression of Nrf2 in PRP group was increased(1.11±0.23),and the protein expression of Keap1 was decreased(1.27±0.15),and

关 键 词：富血小板血浆 脊髓损伤 Nrf2信号通路 

分 类 号：R651.2[医药卫生—外科学]