基于PI3K/AKT/Caspase-9通路研究富氢水在急性放射性脑损伤中的作用及机制  

Effect of hydrogen-rich water on brain injury in rats with acute radiation based on PI3K/AKT/Caspase-9 signaling pathway

作　　者：王永 陈小曼 刘梦雅[2] 罗浩 王永丽[2] 刘晓明[2] 秦秀军[2] WANG Yong;CHEN Xiaoman;LIU Mengya;LUO Hao;WANG Yongli;LIU Xiaoming;QIN Xiujun(School of Forensics,Shanxi Medical University,Taiyuan,Shanxi 030001,China;Shanxi Key Laboratory for Pharmaceutical Toxicology&Radiation Injury Pharmaceuticals/CNNC Key Laboratory for Radiotoxicology and Preclinical Assessment of Radiopharmaceuticals,Department of Radiology and Environmental Medicine,China Institute for Radiation Protection,Taiyuan,Shanxi 030006,China)

机构地区：[1]山西医科大学法医学院,山西太原030001 [2]中国辐射防护研究院放射医学与环境医学研究所,药物毒理与放射损伤药物山西省重点实验室/中核放射毒理与放射性药物临床前评价重点实验室,山西太原030006

出　　处：《环境与职业医学》2023年第3期246-253,共8页Journal of Environmental and Occupational Medicine

基　　金：山西省自然科学基金项目:(201801D121363,202203021211005)。

摘　　要：[背景]目前,放射治疗广泛应用于临床肿瘤的治疗,射线在杀伤肿瘤细胞的同时也会损伤正常组织。有研究表明氢是一种具有前景的治疗相关疾病的方法。[目的]探究磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)/半胱氨酸天冬氨酸特异性蛋白酶9(Caspase-9)信号通路在富氢水对急性放射性脑损伤中的作用及机制。[方法]将40只雄性SD大鼠随机分为对照组、单纯照射(IR)组、高剂量富氢水干预(IR+HHRW)组、低剂量富氢水干预(IR+LHRW)组,每组10只。除对照组外,其余组动物均接受单次20 Gy全脑照射。各组动物均在照射前3 d至照射后7 d内,每天灌胃1次,对照组和IR组给予纯净水(20 mL·kg^(−1)),IR+HHRW组和IR+LHRW组给予富氢水(20 mL·kg^(−1)、10 mL·kg^(−1))。照射7 d后每组各取5只大鼠进行Morris水迷宫实验检测行为学变化;剩余动物麻醉后剖杀、采血、取脑组织检测血液指标变化,进行TUNEL染色观察神经细胞凋亡情况,苏木精和伊红(HE)染色观察组织病理学变化,酶联免疫吸附试验(ELISA)检测氧化应激指标的变化,采用实时荧光定量PCR和Western blotting法检测PI3K、AKT、细胞色素C(Cytc)、Caspase-9的mRNA和蛋白的表达。[结果]照后第7天,与对照组相比,各照射组大鼠体重均降低(P<0.05),并且出现了不同程度的弓背、精神萎靡等症状,IR+HHRW组大鼠较IR组大鼠以上症状明显更轻。行为学检测结果显示:第2~5天IR+HHRW组或IR+LHRW组大鼠相对于IR组逃避潜伏期缩短(P<0.05),且IR+HHRW组大鼠在第6天撤去平台后到达平台原来位置所用时间减少(P<0.05)。血液学检测结果显示,IR组红细胞(RBC)计数、血红蛋白(HGB)量、白细胞(WBC)计数均降低(P<0.05),IR+HHRW组RBC、HGB、WBC指标均有所改善(P<0.05)。HE染色结果显示:IR+HHRW组较IR组的异常神经细胞数量以及碎裂、溶解的细胞核明显减少,损伤程度更轻。氧化应激指标检测结果显示:IR组抑制自由基能[Background]At present,radiation therapy is widely used in clinical treatment of tumors.However,while radiation therapy damages tumor cells,it also injures surrounding normal tissues.Studies have shown that hydrogen is a potential radiation-protective agent.[Objective]To investigate the neuroprotective mechanisms of hydrogen-rich water activating phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)/cysteinyl aspartate specific proteinase-9(Caspase-9)signaling pathway in acute radiation-induced brain injury.[Methods]Forty male SD rats were randomly divided into four groups:control group,irradiation only group(IR),high-dose hydrogenrich water intervention group(IR+HHRW),and low-dose hydrogen-rich water intervention group(IR+LHRW),10 rats in each group.Except for the control group,animals in each group received a single 20 Gy whole brain irradiation.Animals in all groups were gavaged once a day from 3 d before irradiation to 7 d after irradiation,pure water(20 mL·kg^(−1))was given to the control and the IR groups,and hydrogenrich water(20 mL·kg^(−1),10 mL·kg^(−1))was given to the IR+HHRW and the IR+LHRW groups.After 7 d of intervention,5 rats in each group were selected for the Morris water maze experiment for behavioral evaluation.Autopsies were conducted after anesthesia for the remaining animals and blood samples were collected for hematological analysis.Rat brains were harvested for TUNEL staining to observe neuronal apoptosis.HE staining was performed to observe histopathological changes,enzyme-linked immunosorbent assay was adopted to detect oxidative stress-related indicators,and real-time PCR and Western blotting were used to measure the expressions of PI3K/AKT/Caspase-9 pathway-related genes and proteins.[Results]The body weight of rats receiving irradiation decreased after 7 d of irradiation compared with the control group(P<0.05),and the symptoms such as arched back and malaise occurred to varying degrees,and the symptoms of rats in the IR+HHRW group were significantly milder than those in the I

关 键 词：富氢水 电离辐射 磷脂酰肌醇3-激酶/蛋白激酶B/半胱氨酸天冬氨酸特异性蛋白酶9信号通路 神经细胞凋亡 

分 类 号：R11[医药卫生—公共卫生与预防医学]