人参皂苷Rg1在小鼠创伤性脑损伤修复中的作用  

Repairing Effects of Ginsenoside Rg1 on Traumatic Brain Injury in Mice

作　　者：郭文文 赵亚 王颖花 刘可 葛煦 张延英 汪永锋 师长宏 GUO Wenwen;ZHAO Ya;WANG Yinghua;LIU Ke;GE Xu;ZHANG Yanying;WANG Yongfeng;SHI Changhong(Basic Medical College,Gansu University of Traditional Chinese Medicine,Lanzhou 730030,China;Laboratory Animal Center,Air Force Military Medical University,Xi'an 710032,China;Gansu Province Experimental Animal Industry Technology Center,Lanzhou 730030,China)

机构地区：[1]甘肃中医药大学基础医学院,兰州730030 [2]空军军医大学实验动物中心,西安710032 [3]甘肃省实验动物行业技术中心,兰州730030

出　　处：《实验动物与比较医学》2023年第3期243-252,共10页Laboratory Animal and Comparative Medicine

基　　金：陕西省创新能力支撑计划-科技资源开放共享平台项目“基于诱导性多潜能干细胞的人神经元嵌合小鼠的制备与评价”(2021PT037);军队实验动物专项科研课题“人神经元嵌合小鼠的制备与评价”(SYDW〔2018〕01)。

摘　　要：目的探讨人参皂苷Rg1在创伤性脑损伤(traumatic brain injury,TBI)小鼠模型血脑屏障、神经炎症、行为学功能等方面的作用。方法实验分2部分。第一部分是将27只SPF级雄性BALB/c小鼠随机分为空白组、假手术组、TBI模型组,每组9只;TBI模型组开颅后采用受控皮质冲击(controlled cortical impact,CCI)方式造模,假手术组只开颅不进行打击,空白组不经任何处理;手术后进行造模效果评价。第二部分是将40只雄性BALB/c小鼠随机分为假手术组、3个剂量的人参皂苷Rg1治疗组和溶剂DMSO对照组,每组8只小鼠。Rg1治疗组在TBI建模成功后6 h腹腔注射剂量分别为10、20、40 mg/kg的人参皂苷Rg1,而DMSO对照组给予等量的1%DMSO,持续给药1周,每天2次。于造模后1、3、7、14 d分别进行改良的神经损伤严重程度评分(modified neurological severity scores,mNSS);取造模后第3天的小鼠脑组织,采用蛋白质印迹法检测血脑屏障渗漏情况;第14、16天分别采用高架十字迷宫实验、水迷宫实验检测小鼠神经行为功能;第28天麻醉、灌注后取脑,免疫荧光染色观察小胶质细胞、星形胶质细胞活化等神经炎症。结果人参皂苷Rg1治疗组的血脑屏障标志物MMP-9表达量减少(P<0.01),小胶质细胞(Iba-1阳性表达)和星形胶质细胞(GFAP阳性表达)数量均明显减少(P<0.05),提示神经炎症得到抑制,且以20 mg/kg剂量效果最好(P<0.01)。人参皂苷Rg1治疗组小鼠的mNSS评分显著低于溶剂DMSO对照组(P<0.01),进入开放臂次数比例显著高于DMSO对照组(P<0.05);其在水迷宫实验平台所在象限的时间比及穿越平台的次数均显著多于DMSO对照组(P<0.05),且均以20 mg/kg剂量效果最佳。结论成功构建TBI小鼠模型并用于人参皂苷Rg1的损伤修复研究。人参皂苷Rg1能够显著改善TBI模型小鼠血脑屏障,减轻神经炎症,发挥改善神经行为功能的作用,且以20 mg/kg剂量作用效果最为显著。Objective To explore the effects of ginsenoside Rg1 on blood-brain barrier,neuroinflammation and behavioral function of traumatic brain injury(TBI)mouse model.Methods The experiment was divided into two parts.In the first part,27 SPF male BALB/c mice were randomly divided into blank group,sham operation group and TBI model group,with 9 mice in each group.TBI model group was made by controlled cortical impact(CCI)after craniotomy,while sham operation group was only performed craniotomy without any treatment,and the blank group was not treated at all.The effect of modeling was evaluated after operation.In the second part,50 male BALB/c mice were randomly divided into sham operation group,three different drug dosage groups and solvent(DMSO)control group,with 8 mice in each group.The drug treatment groups were injected with ginsenoside Rg1 at the doses of 10,20 and 40 mg/kg respectively 6 hours after TBI model had been successfully established,while the DMSO control group was given the same amount of 1%DMSO for one week,twice a day.Modified neurological severity scores(mNSS)were performed on the 1st,3rd,7th and 14th day after modeling,and the blood-brain barrier leakage was detected by Western blotting on the 3rd day after modeling.On the 14th and 16th day,the elevated cross maze test and water maze test were used to detect the neurobehavioral function.On the 28th day after anesthesia and perfusion,the brains were taken out,and the neuroinflammation such as activation of microglia and astrocytes was observed by immunofluorescence staining.Results The expression level of MMP-9,a marker of blood-brain barrier,decreased in ginsenoside Rg1 treatment group(P<0.01).The number of microglia(Iba-1 positive)and astrocyte(GFAP positive)cells decreased significantly(P<0.05),which indicated that neuroinflammation was inhibited,and the best effect was achieved at the dosage of 20 mg/kg(P<0.01).The mNSS of mice in ginsenoside Rg1 treatment group were significantly lower than those in DMSO control group(P<0.01),and the proportion of

关 键 词：创伤性脑损伤 人参皂苷RG1 小鼠 神经炎症 行为学 

分 类 号：R-332[医药卫生] Q95-33[生物学—动物学]