富氢液对化疗所致小鼠认知功能障碍的影响及机制  

Role of HRS in amelioration of chemotherapy-induced cognitive impairment in mice

作　　者：胡南 赵洪伟 张霄蓓 王捷夫[1,2,3] HU Nan;ZHAO Hongwei;ZHANG Xiaobei;WANG Jiefu(Tianjin Medical University Cancer Institute and Hospital,National Clinical Research Center for Cancer,Tianjing 300060,China)

机构地区：[1]天津医科大学肿瘤医院国家肿瘤临床研究中心,天津300060 [2]天津市肿瘤防治重点实验室 [3]天津市恶性肿瘤临床医学研究中心

出　　处：《山东医药》2019年第26期13-17,共5页Shandong Medical Journal

基　　金：国家自然科学基金青年科学基金项目(81701081)

摘　　要：目的观察富氢液对化疗所致小鼠认知功能障碍的影响,并探讨其可能的机制。方法将8周成年CD1小鼠随机分为对照组(C组)、富氢液组(HRS组)、5-氟尿嘧啶组(5-FU组)、5-FU+富氢液组(5-FU+HRS)和5-FU+富氢液+Sirtuin-1抑制剂Nicotinamide组(5-FU+HRS+Nico组)各54只,HRS组和5-FU+HRS组、5-FU+HRS+Nico组腹腔注射2%富氢液(5 mL/kg,连续注射14 d),5-FU+HRS+Nico组予腹腔注射富氢液后即予Nicotinamide注射,5-FU组、5-FU+HRS组和5-FU+HRS+Nico组腹腔注射5-FU(20 mg/kg,连续注射14 d)。每组分别于处理后24 h,采用条件恐惧实验(5 min内的僵直时间)以及Y迷宫实验(5 min内的N臂停留时间)判断认知功能;处死小鼠并取脑,采用免疫荧光法测算海马组织巨噬细胞的分型面积及所占比例(CD11b和CD206分别标记M1型与M2型巨噬细胞,Iba-1标记巨噬细胞),采用ELISA法检测海马组织中的M1型巨噬细胞因子(IL-6、IL-1β)与M2型巨噬细胞因子(IL-10),采用Western blotting法检测海马组织中的Sirtuin-1和细胞内Notch受体结构域(NICD)总蛋白和核蛋白。结果与C组比较,5-FU组、5-FU+HRS组和5-FU+HRS+Nico组小鼠5 min内的僵直时间、N臂停留时间缩短,海马组织中M1型巨噬细胞面积增大、M2型巨噬细胞面积减小,海马组织中IL-1β、IL-6表达上升而IL-10表达下降,海马组织中Sirtuin-1总蛋白和核蛋白表达均下降而NICD总蛋白和核蛋白表达均升高(P均<0.05),HRS组各指标差异无统计学意义(P均>0.05);与5-FU组比较,5-FU+HRS组小鼠5 min内的僵直时间、N臂停留时间延长,海马组织中M1型巨噬细胞面积减小、M2型巨噬细胞面积增大,海马组织中IL-1β、IL-6表达下降而IL-10表达上升,海马组织中Sirtuin-1总蛋白和核蛋白表达均升高而NICD总蛋白和核蛋白表达均下降(P均<0.05),5-FU+HRS+Nico组各指标差异无统计学意义(P均>0.05)。结论2%富氢液可减轻5-FU所致的小鼠认知功能障碍;其机制与通过上调Sirtuin-1抑制Notch通路激Objective To evaluate the role of hydrogen-rich saline(HRS)in mice with cognitive impairment induced by chemotherapy and to explore its underlying mechanism.Methods Adult CD1 mice(8 weeks old)were randomly divided into 5 groups:the control group(group C),HRS group,5-fluorouracil group(5-FU group),5-FU+HRS group(group 5-FU+HRS)and 5-FU+HRS+Sirtuin-1 antagonist Nicotinamide group(group 5-FU+HRS+Nico).Mice in the group HRS,5-FU+HRS and 5-FU+HRS+Nico received HRS(2%,5 mL/kg)peritoneal perfusion continuously for 14 days.Mice in the group 5-FU+HRS+Nico received Nicotinamide(30 mg/kg)peritoneal perfusion immediately after HRS perfusion each time.Mice in the group 5-FU,group 5-FU+HRS and 5-FU+HRS+Nico reveiced 5-FU(20 mg/kg)peritoneal perfusion before they were given HRS for 14 days continuously.At 24 hours after treatment,the cognitive function was judged by conditional fear test(freezing time within 5 minutes)and Y maze test(new arm residence time).Brain tissues were harvested at 24 h after treatment for determining Notch intracellular domain(NICD),Sirtuin-1 and macrophage type.The classification of macrophage in hippocampi was measured by immunofluorescence(CD11b and CD206 labeled M1 and M2,and Iba-1 labeled macrophages).M1 macrophage cytokine(IL-6 and IL-1β)and M2 macrophage cytokine(IL-10)in hippocampi was detected by ELISA.Sirtuin-1 protein and NICD total protein and nucleoprotein in hippocampi was detected by Western blotting.Results Compared with the group C,the ratio of freezing time and new arm residence time decreased significantly in the 5-FU,5-FU+HRS and 5-FU+HRS+Nico groups(both P<0.05).The area and ratio of M1 type macrophage increased significantly,and those of M2 type decreased in the 5-FU,5-FU+HRS and 5-FU+HRS+Nico groups(all P<0.05).The expression of IL-1βand IL-6 in the hippocampi increased and the expression of IL-10 decreased in the 5-FU,5-FU+HRS and 5-FU+HRS+Nico groups(all P<0.05).The expression of Sirtuin-1 decreased and NICD(total and nuclear protein)increased in the 5-FU,5-FU+HRS and 5-FU+HRS

关 键 词：巨噬细胞极化 组蛋白去乙酰化酶 NOTCH通路 化学治疗 认知功能障碍 小鼠 

分 类 号：R741.05[医药卫生—神经病学与精神病学]