三七和PDTC对创伤性脑损伤大鼠自噬机制的干预作用  

Effect of Panax notoginseng and Pyrrolidinedithiocarbamate on autophagy in rats with traumatic brain injury

作　　者：石莹[1] 王灵聪[1] 周霞庆 蒋慧芳[2] SHI Ying;WANG Lingcong;ZHOU Xiaqing;JIANG Huifang(Department of Intensive Care Unit,the First Affiliated Hospital,Zhejiang Chinese Medical University,Hangzhou 310006,China)

机构地区：[1]浙江中医药大学附属第一医院重症医学科,杭州310006 [2]浙江省立同德医院血液科

出　　处：《浙江医学》2021年第7期700-704,I0004,共6页Zhejiang Medical Journal

基　　金：浙江省自然科学基金(LQY19H080001);浙江省教育厅一般科研项目(Y202045214)。

摘　　要：目的探讨三七和吡咯醛二硫氨基甲酸酯(PDTC)对创伤性脑损伤(TBI)大鼠自噬机制的干预作用。方法将36只SD成年雄性大鼠按随机数字表法分为假手术组(SHAM组)、模型组(M组)、三七组(PN组)、NF-κB抑制剂PDTC组(PDCT组)、三七+PDTC组(PN+PDCT组)、三七+氯喹组(PN+CHQ组),每组6只。SHAM组大鼠进行手术及麻醉,但无冲击损伤过程,其余各组均采用改良的Feeney自由落体模型制备法建立TBI模型。6组大鼠术前3 d、术后3 d予0.9%氯化钠注射液1 ml每天分2次灌胃,术前1 d、术后3d予0.9%氯化钠注射液1 m1腹腔注射,1次/d。PN组灌胃时加入三七粉2.5 g/kg。PDTC组腹腔注射时加入PDTC 120 mg/kg。PN+PDTC组灌胃时加入三七粉2.5 g/kg,腹腔注射时加入PDTC 120 mg/kg。PN+CHQ组灌胃时加入三七粉2.5 g/kg,腹腔注射时加入氯喹60 mg/kg。术后第4天乙醚麻醉处死,取大脑组织进行检测。观察各组大鼠造模后12、24、72 h神经功能缺陷评分,大鼠脑组织HE染色、Nis sl染色观察脑组织损伤情况,透射电镜观察细胞自噬体,Wes tern blot检测各组大鼠脑组织泛素结合蛋白p62、Beclinl和微管相关蛋白轻链3(LC3)表达。结果M组大鼠各时间点神经功能缺陷评分明显下降,HE染色和Nissl染色脑组织损伤明显,透射电镜下脑细胞中可见大量自噬体,自噬蛋白明显升高(P<0.05或0.01)。PN组、PDTC组、PN+PDTC组、PN+CHQ组均能改善72 h神经功能缺陷评分,同时HE染色和Nissl染色脑组织损伤较M组改善,透射电镜下脑细胞中可见自噬体较M组减少,p62、Beclinl和LC3较M组均明显下降(P<0.05或0.01)。PN组、PDTC组、PN+PDTC组、PN+CHQ组组间比较差异均无统计学意义(均P>0.05)。结论TBI大鼠存在自噬并伴有脑功能损伤,三七以及PDTC均能干预TBI大鼠自噬过程,促进脑功能恢复,三七与PDTC以及三七与氯喹并无明显协同作用。Objective To investigate the effects of panax notoginseng(PN)and pyrrolidinedithiocarbamate(PDTC)on autophagy in rats with traumatic brain injury.Methods Thirty-Six Sprague-Dawley(SD)rats were randomly divided into six groups:sham group,model group,Panax notoginseng(PN)group,PDTC group,PN+PDTC group,PN+chloroquine(CHQ)group,with 6 rats in each group.The traumatic brain injury was induced by modified Feeney method in model group and 4 treatment groups,and the sham operation was given to rats in sham group.In the PN group,2.5 g/kg of PN powder was given by gavage,in the PDTC group 120 mg/kg of PDTC was given by intraperitoneal injection,in the PN+PDTC group both PN powder and PDTC were given as above,in the PN+CHQ group,2.5 g/kg of PN powder was given by gavage,and 60 mg/kg of CHQ was given by intra peritoneal injection.The neural function defrct score of rats in each group were ob served at 12 h,24 h and 72 h.After 4 days the rats were sacrificed and the brain was taken for examination.The brain tissue damage was observed by HE staining and Nissl staining,the autophagy was observed by transmission electron microscopy,and the expressions of autophagy-related proteins p62,Beclinl and LC3 in the brain tissues were detected by Western blot Results In the model group,the neural function defectscores at all time points were significantly decreased,HE staining and Nissl staining showed significant brain tissue damage,transmission electron microscopy revealed a large number of autophagosomes,and the expressions of autophagy proteins in brain tissues were significantly elevated(P<0.05 or 0.01).Compared with the model group,the neural function defect score of rats at 72 h was improved,the brain tissue damage was attenuated,autophagosomes were reduced as shown by transmission electron microscopy and the expressions of autophagy proteins were significantly decreased in the PN group,PDTC group,PN+PDTC group,PN+CHQ group.And there were no significant differences in above changes among the PN group,PDTC group,PN+PDTC group,PN+CH

关 键 词：脑损伤 自噬 三七 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]