摘要
目的探讨胰高血糖素样肽-1受体激动剂Exendin-4对颅脑创伤小鼠神经功能的影响。方法 72只雄性C57BL/6J小鼠随机分为假手术组、颅脑创伤组、治疗组各24只。颅脑创伤组、治疗组采用脑皮质打击致伤法制备颅脑创伤模型,假手术组仅打开骨窗但不打击脑皮质。治疗组于打击后即刻腹腔注射Exendin-4 20μg/kg,之后每隔24 h给药1次,共给药3次;假手术组和颅脑创伤组腹腔注射等量生理盐水。每组随机选取12只小鼠,于打击前和打击后第3、5和7天行神经功能缺陷评分(neurological severity score, NSS)、足失误试验和横木行走试验(beam walking test, BWT)。每组剩余12只小鼠于打击后第3天处死,采用FJC染色法检测脑组织神经元变性坏死,采用免疫荧光法检测胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)和离子钙接头蛋白-1(ionized calcium-binding adapter molecule-1, Iba-1)阳性表达,采用ELISA法检测肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)和白细胞介素-1β(interleukin-1β, IL-1β)表达。结果打击后第3、5、7天,NSS在颅脑创伤组[(8.00±1.28)、(6.25±1.36)、(6.00±1.41)分]和治疗组[(5.58±1.00)、(4.17±1.19)、(2.42±1.31)分]均高于假手术组[(1.50±0.90)、(1.33±0.89)、(1.25±0.97)分](P<0.05),足失误率在颅脑创伤组[(27.67±5.10)%、(19.83±3.13)%、(16.83±3.46)%]和治疗组[(20.00±4.09)%、(15.17±1.34)%、(10.33±1.87)%]均高于假手术组[(5.17±1.80)%、(4.75±2.30)%、(5.00±2.34)%],BWT评分在颅脑创伤组[(4.17±0.83)、(3.75±0.45)、(2.92±0.51)分]和治疗组[(3.25±0.45)、(2.25±0.75)、(1.25±0.45)分]均高于假手术组[(0.45±0.25)、(0.39±0.17)、(0.45±0.25)分](P<0.05),颅脑创伤组各时间点NSS、足失误率、BWT评分高于治疗组(P<0.05)。打击后第3天,颅脑创伤组脑组织FJC阳性细胞率[(36.19±10.29)%]高于治疗组[(14.62±3.98)%]和假手术组[(9.48±2.14)%](P<0.05),治疗组与假手术组比较差异无统计学意义(P>0.05);颅脑创伤组和治疗组脑组织GFAP阳性细胞率[(30.77±6.09)%、(13.75±8.09)%]、Iba-1阳性细胞率[(32.21±12.48)%、(16.06±4.27)%]、TNF-α表达[(112.48±6.67)、(65.81±13.52)ng/L]、IL-1β表达[(91.25±13.24)、(51.43±5.48)ng/L]高于假手术组[(6.31±2.20)%、(9.35±3.76)%、(38.02±1.85)ng/L、(31.21±6.99)ng/L](P<0.05),颅脑创伤组高于治疗组(P<0.05)。结论腹腔注射Exendin-4可改善颅脑创伤小鼠神经运动和协调功能,减轻神经功能损伤,其作用机制可能与抑制神经胶质细胞激活、减少炎性因子释放有关。
Objective To investigate the effect of glucagon-like peptide-1 receptor agonist exendin-4 on the neurological function after traumatic brain injury(TBI)in mice.Methods Seventy-two male C57 BL/6 J mice were randomly divided into sham-operation group,TBI group and therapy group,with 24 mice in each group.In TBI group and therapy group,TBI models were established by the cortical contusion impact methods,and sham-operation group was only opened the bone window but no cortical contusion impact.Therapy group was intraperitoneally injected with exendin-4(20μg/kg)immediately after modeling,followed by once every 24 h,totally 3 times,and TBI and sham-operation groups were intraperitoneally injected with the equivalent volume of normal saline.Twelve mice per group were randomly selected and orderly determined the neurological severity score,foot fault test,and beam walking test before modeling as well as on the 3 rd,5 th,and 7 th days after modeling,respectively.The other 12 mice in each group were sacrificed on the 3 rd day after modeling,the neuronal degeneration was detected by FJC,the positive expressions of glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1 levels were detected by Western blot,and the levels of tumor necrosis factor-αand interleukin-1βwere detected by ELISA.Results On the 3 rd,5 th and 7 th days after modeling,the neurological severity scores were higher in TBI group(8.00±1.28,6.25±1.36,6.00±1.41)and therapy group(5.58±1.00,4.17±1.19,2.42±1.31)than those in sham-operation group(1.50±0.90,1.33±0.89,1.25±0.97)(P<0.05),the incidences of foot faults were higher in TBI group(27.67±5.10)%,(19.83±3.13)%,(16.83±3.46)%)and therapy group(20.00±4.09)%,(15.17±1.34)%,(10.33±1.87)%)than those in sham-operation group(5.17±1.80)%,(4.75±2.30)%,(5.00±2.34)%)(P<0.05),the beam walking test scores were higher in TBI group(4.17±0.83,3.75±0.45,2.92±0.51)and therapy group(3.25±0.45,2.25±0.75,1.25±0.45)than those in sham-operation group(0.45±0.25,0.39±0.17,0.45±0.25)(P<0.05),and all above indexes were higher in TBI group than those in therapy group(P<0.05).On the 3 rd day after modeling,the FJC-positive rate was higher in TBI group(36.19±10.29)%)than that in therapy group(14.62±3.98)%)and sham-operation group(9.48±2.14)%)(P<0.05),and showed no significant difference between therapy group and sham-operation group(P>0.05).The glial fibrillary acidic protein positive rate,ionized calcium-binding adapter molecule 1 positive rate,tumor necrosis factor-αlevel and interleuin-1βlevel were the highest in TBI group(30.77±6.09)%,(32.21±12.48)%,(112.48±6.67)ng/L,(91.25±13.24)ng/L,followed by therapy group(13.75±8.09)%,(16.06±4.27)%,(65.81±13.52)ng/L,(51.43±5.48)ng/L and sham-operation group(6.31±2.20)%,(9.35±3.76)%,(38.02±1.85)ng/L,(31.21±6.99)ng/L in turn(P<0.05).Conclusion Intraperitoneal injection of exendin-4 could greatly improve the neurological performances and cognitive function,and attenuate the neural degeneration,probably by inhibiting glial cells activation and suppressing the inflammatory cytokines release.
作者
曹义坡
姚旻
刁云锋
董铮
吕晓楠
李红涛
CAO Yipo;YAO Min;DIAO Yunfeng;DONG Zheng;LYU Xiaonan;LI Hongtao(Department of Endocrinology,Hospital of Tianjin Armed Police Corps,Tianjin 300162,China;Institute of Neurological Trauma and Repair,Armed Police Characteristic Medical Center,Tianjin 300162,China)
出处
《中华实用诊断与治疗杂志》
2020年第7期660-665,共6页
Journal of Chinese Practical Diagnosis and Therapy
基金
吴阶平医学基金增补项目(20151012)。
