甘珀酸治疗胶原酶诱导的大鼠丘脑出血性卒中后疼痛的相关机制  

Carbenoxolone alleviates hemorrhage-induced thalamic pain induced by collagenase of rats

作　　者：李海洋 赖生龙 苗壮 梁庆华 李玉 Li Haiyang;Lai Shenglong;Miao Zhuang;Liang Qinghua;Li Yu(Department of neurosurgery,People’s hospital of Henan province,People’s hospital of ZhengZhou University,School of Clinical Medicine,Henan University,ZhengZhou 450003,China)

机构地区：[1]河南省人民医院(郑州大学人民医院河南大学人民医院)神经外科,郑州450003

出　　处：《中华实验外科杂志》2023年第11期2265-2269,共5页Chinese Journal of Experimental Surgery

摘　　要：目的:利用在大鼠丘脑部位注射Ⅳ型胶原酶诱导出血性脑卒中导致慢性卒中后疼痛(CPSP)大鼠模型,并在CPSP大鼠脑室内注射甘珀酸观察对卒中后疼痛大鼠是否有镇痛作用及探索相关机制。方法:为检测模型是否成功及大鼠行为学,将SD大鼠采用随机数表法分为对照组(丘脑后外侧注射生理盐水,每组5只)和实验组(丘脑后外侧注射胶原酶,每组5只)。实验组脑立体定位注射Ⅳ胶原酶0.125 U/0.5μl,术后观察大鼠的运动功能,应用Von-Frey丝、红外线热痛仪检测大鼠痛觉的变化。为检测甘珀酸作用将大鼠随机分为对照组(丘脑后外侧注射生理盐水,每组13只);CPSP组(丘脑后外侧注射胶原酶,每组13只)、CPSP+甘珀酸组(丘脑后外侧注射胶原酶,第7天侧脑室注射甘氨酸,每组13只);其中各5只检测行为学,各5只检测胶质纤维酸性蛋白(GFAP),各3只检测免疫荧光。取大鼠的丘脑检测GFAP的表达变化。在CPSP大鼠脑室内注射甘珀酸20μg(浓度2 g/L),观察甘珀酸对CPSP的影响。多组间比较采用单因素方差分析或双因素方差分析。结果:大鼠丘脑腹后区注射胶原酶后,大鼠的机械痛阈值于胶原酶注射后第7天从(23.65±3.03)g降低到(6.45±1.12)g,差异有统计学意义(F=13.010,P<0.05),持续28 d,热痛阈值从(12.50±0.75)s降低到(6.71±0.88)s,差异有统计学意义(F=10.362,P<0.05),但大鼠的运动功能不受影响。通过脑室注射给予甘珀酸后第2天,CPSP大鼠的机械痛阈值回升到(16.65±3.24)g,热痛阈值回升到(9.63±1.62)s,差异有统计学意义(F=16.161,P<0.05)。CPSP组大鼠术后第7天背侧丘脑GFAP表达升高(3.20±0.89)倍,给予甘珀酸后背侧丘脑GFAP表达下降到正常值的(1.40±0.31)倍,差异有统计学意义(F=18.380,P<0.05)。结论:出血性脑卒中后的肢体疼痛可能与出血区周围星形胶质细胞的活化有关,给予星形胶质细胞的抑制剂甘珀酸后可以缓解CPSP。Objective To investigate the mechanisms of Carbenoxolone treatment for collagenase-induced chronic post-stroke pain(CPSP)in the thalamus of rats.Methods In order to test the success of the model and the behavior of SD rats,the male SD rats were randomly divided into the sham surgery control group and the CPSP group(5 rats per group).In the CPSP group,a stereotactic injection of CollagenaseⅣ(0.125 U/0.5μl,dissolved in saline)was administered into the brain.Behaviors were measured by Von-Frey hairs and thermal test apparatus.Carbenoxolone(20μg,2 g/L)is administrated into ventricle to detect the behavior changes of CPSP rats.To investigate the effects of gabapentin,rats were randomly divided into three groups:control group,CPSP group,and CPSP plus gabapentin group.There were 13 rats in each group,with 5 rats for behavior analysis,5 rats for glial fibrillary acidic protein(GFAP)detection,and 3 rats for immunofluorescence analysis.Western-blot were applied to detect the expression of GFAP in thalamus.Immunostaining was used to detect Astrocytes in thalamus.One-way analysis of variance or two-way analysis of variance was used for intergroup comparisons.Results At 7 d after microinjection of collagenaseⅣinto thalamus of rats,the mechanical threshold decreased from(23.65±3.03)g to(6.45±1.12)g(F=13.010,P<0.05)and the heat threshold decreased from(12.50±0.75)s to(6.71±0.88)s(F=10.362,P<0.05),and this persisted for 28 days,but the motor function of rats was not affected.The mechanical threshold and thermal pain threshold of CPSP rats were reversed after intraventricular injection of Carbenoxolone.The expression of GFAP in thalamus of CPSP rats increased to(3.20±0.89)at 7 d,but it decreased to(1.40±0.31)times the normal value in the dorsal thalamus after administration with Carbenoxolone,and the difference was statistically significant(F=18.380,P<0.05).Conclusion Activation of astrocytes may be related to CPSP induced by hemorrhagic-stroke,and the inhibitor of astrocyte can alleviate CPSP.

关 键 词：出血性脑卒中 卒中后疼痛 星形胶质细胞 甘珀酸 

分 类 号：R743.3[医药卫生—神经病学与精神病学]