基于调控核因子E2相关因子2探讨蟾酥对脑缺血再灌注损伤小鼠星形胶质细胞的影响  

Protective effect of Chansu preconditioning on cerebral ischemia-reperfusion injury in mice by regulating the expression of nuclear factor E2-related factor 2 in astrocyte

作　　者：钱程[1] 高振 陈卓[3] QIAN Cheng;GAO Zhen;CHEN Zhuo(Department of Medical,The Second Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001;Department of Radiotherapy,The Second Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001;Department of Intervention,The Second Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001)

机构地区：[1]南通大学第二附属医院医务科,江苏南通226001 [2]南通大学第二附属医院放疗科,江苏南通226001 [3]南通大学第二附属医院介入科,江苏南通226001

出　　处：《河北中医》2022年第12期2047-2051,共5页Hebei Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81701286)。

摘　　要：目的研究蟾酥预处理通过调控星形胶质细胞核因子E2相关因子2(Nrf2)表达对脑缺血再灌注损伤小鼠的保护作用。方法将60只C57小鼠随机分为假手术组、模型组和蟾酥注射液低、中、高剂量组,每组12只。模型组采用线栓法制备脑缺血再灌注模型;假手术组血管分离后不插入线栓,结扎消毒后缝合皮下组织及皮肤即可,其余操作同模型组;蟾酥注射液低、中、高剂量组分别于缺血前30 min经腹腔注射蟾酥注射液1.6、3.2、6.4 mL/kg,其余操作同模型组。再灌注后24 h,采用Longa’s评分法评定各组小鼠神经功能。再灌注后72 h以2,3,5-氯化三苯基四氮唑(TTC)染色法检测小鼠脑梗死体积百分比,蛋白免疫印迹法检测小鼠脑组织胶质纤维酸性蛋白(GFAP)和Nrf2蛋白表达水平,免疫荧光法检测小鼠脑组织GFAP和Nrf2阳性表达数。结果与假手术组比较,模型组小鼠Longa’s评分、脑梗死体积百分比、GFAP蛋白表达和阳性细胞数均显著提高(P<0.05),Nrf2蛋白表达和阳性细胞数均降低(P<0.05);与模型组比较,蟾酥注射液低、中、高剂量组小鼠Longa’s评分、脑梗死体积百分比、GFAP蛋白表达和阳性细胞数均降低(P<0.05),Nrf2蛋白表达和阳性细胞数均升高(P<0.05)。结论蟾酥预处理对脑缺血再灌注损伤小鼠具有保护作用,其保护机制可能是通过抑制脑组织星形胶质细胞GFAP和促进Nrf2的表达来实现的。Objective To study the protective effect of Chansu preconditioning on cerebral ischemia-reperfusion injury in mice by regulating the expression of nuclear factor E2-related factor 2(Nrf2)in astrocyte.Methods Sixty C57 mouse were randomly assigned into the sham operation group,the model group,and the low,medium-,high-dose Chansu injection group,with 12 in each group.The mouse in the model group were prepared with suture-occluded method to prepare cerebral chemia-reperfusion model,the mouse in the sham operation group weren’t inserted with sutures after blood vessel separation.Only the subcutaneous tissue and skin were sutured after ligation and disinfection,and the remaining operations were the same to those for the mouse in the model group.The mouse in the low,medium-,high-dose Chansu group were injected intra-peritoneally with 1.6,3.2,and 6.4 mL/kg of Chansu injection 30 minutes before ischemia,respectively,and other operations were the same to those for the model group.The neurological function of mouse in groups after reperfusion for 24 hours was evaluated by Longa’s score.After 72 hours of reperfusion,the 2,3,5-triphenyltetrazolium chloride(TTC)staining was performed to detect cerebral infarction volume.Western blotting was conducted to assess the protein expression levels of glial fibrillary acidic protein(GFAP)and Nrf2.The immunofluorescence method was used to test the number of positive expressions of GFAP and Nrf2 in mouse brain tissue.Results Compared with the sham operation group,the Longa’s score,the percentage of cerebral infarction volume,the protein expression and the number of positive cells of GFAP in the model group were significantly increased(P<0.05),and the expression and the number of positive cells of Nrf2 protein were significantly decreased(P<0.05);Compared with the model group,the Longa’s score,the percentage of cerebral infarction volume,the protein expression and the number of positive cells of GFAP in Chansu injection groups were significantly decreased(P<0.05),and the protein

关 键 词：再灌注损伤 蟾酥 小鼠 神经保护 星形细胞 细胞核因子E2相关因子2 动物实验 

分 类 号：R743.9[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]