6-羟基染料木素治疗高原缺氧小鼠肺损伤的作用机制  

Mechanisms of 6-Hydroxygen Genistein in the Treatment of Pulmonary Injury in High-Altitude Hypoxic Mice

作　　者：马川 王小娟 杨陈煜 张书瑜 杨宝乐 景临林 马慧萍 MA Chuan;WANG Xiaojuan;YANG Chenyu;ZHANG Shuyu;YANG Baole;JING Linlin;MA Huiping(College of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730030,China;Department of Pharmacy,The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army,Lanzhou 730050,China;Department of Pharmacy,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 7100612,China)

机构地区：[1]甘肃中医药大学药学院,兰州730030 [2]中国人民解放军联勤保障部队第九四〇医院药剂科,兰州730050 [3]西安交通大学第一附属医院药学部,西安7100612

出　　处：《四川大学学报(医学版)》2025年第1期222-229,共8页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81571847、No.81872796);军队后勤科研计划项目(No.2023HQZZ-02);联勤保障部队第九四〇医院专项培育项目(No.2021yxky015);西北中藏药省部共建协同创新中心2022年度开放基金项目(No.Xbzzy-2022-04)资助。

摘　　要：目的探讨6-羟基染料木素(6-hydroxygenistein,6-OHG)治疗高原缺氧诱导肺损伤的作用机制。方法从Swiss Target Prediction、SuperPred、GeneCards和OMIM等数据库中筛选获得6-OHG和高原缺氧肺损伤的交集靶点。运用STRING数据库和Cytoscape软件对药物与疾病的交集靶点构建蛋白互作网络,并将度值大于中位数的作为关键靶点;使用DAVID数据库对关键靶点进行GO富集和KEGG富集分析获得相关信号通路;用Maestro 13.7软件进行分子对接验证。采用大型低压低氧舱建立小鼠高原肺损伤模型,将42只雄性Balb/c小鼠随机分为3组,每组14只:正常对照组(当地海拔1400 m,一次性腹腔注射生理盐水)、模型组(一次性腹腔注射生理盐水)、6-OHG组(一次性腹腔注射100 mg/kg 6-OHG)。给药1 h后,模型组和6-OHG组小鼠置于大型低压低氧模拟动物实验舱中,然后以10 m/s上升至海拔8000 m,并维持24 h,再降至海拔3500 m。处死3组小鼠,取肺组织,测定肺含水量,HE染色观察病理学变化,测定MDA(malondialdehyde)、H_(2)O_(2)、TSOD(total superoxide dismutase)和GSH(glutathione)的水平,Western blot检测p-PI3K/PI3K、p-AKT/AKT、缺氧诱导因子1α(HIF-1α)和VEGF(vascular endothelial growth factor)蛋白表达。结果筛选出丝氨酸/苏氨酸蛋白激酶1(AKT1)、HIF-1α、表皮生长因子受体(EGFR)、基质金属蛋白酶9(MMP9)、过氧化物酶增殖物激活受体A(PPARA)等关键靶点,GO富集和KEGG富集分析显示6-OHG治疗高原缺氧肺损伤的靶点主要参与PI3K/AKT、HIF-1α/VEGF、肿瘤坏死因子(TNF)等信号通路。动物实验结果显示,与模型组相比,6-OHG可以显著改善高原缺氧诱导的肺组织病理损伤,且MDA、H_(2)O_(2)、GSH和T-SOD水平与模型组相比,差异有统计学意义(P<0.01)。Western blot实验结果显示,6-OHG组肺组织p-PI3K/PI3K、p-AKT/AKT、HIF-1α和VEGF较模型组相比,差异有统计学意义(P<0.01)。分子对接结果表明6-OHG可与PI3K、AKT、HIF-1α和VEGF形�Objective To investigate the mechanisms of 6-hydroxygenistein(6-OHG)in the treatment of highaltitude hypoxia-induced lung injury.Methods The intersection targets of 6-OHG and high-altitude hypoxia-induced lung injury were identified using databases,including Swiss Target Prediction,SuperPred,GeneCards,and OMIM.The STRING database and Cytoscape software were used to construct a protein interaction network for the intersection targets of drugs and diseases,and targets with degree values greater than the median were identified as key targets.GO and KEGG enrichment analyses of key targets were performed using the DAVID database to identify relevant signaling pathways.The Maestro 13.7 software was used for molecular docking validation.A large hypobaric hypoxic chamber was used to establish a high-altitude lung injury model in mice.A total of 42 male BALB/c mice were randomly assigned to 3 groups(n=14 in each group),including a normal control group,which was exposed to the environmental conditions at the altitude of 1400 m and received a single intraperitoneal injection of normal saline,a model group,which received a single intraperitoneal injection of normal saline,and a 6-OHG group,which received a single intraperitoneal injection of 6-OHG at 100 mg/kg.Then,1 h after drug administration,mice in the model and 6-OHG groups were placed in a large hypobaric hypoxic simulation chamber for animal experiments.Then,they ascended to an altitude of 8000 m at a speed of 10 m/s,remained in that environment for 24 h,and then descended to an altitude of 3500 m.Mice in the three groups were sacrificed,and their lung tissues were extracted to measure the water content in the lungs.Pathological changes were observed using HE staining,and the levels of malondialdehyde(MDA),H_(2)O_(2),total superoxide dismutase(T-SOD),and glutathione(GSH)were measured.Western blot was performed to determine the expression levles of p-PI3K/PI3K,p-AKT/AKT,hypoxia-inducible factor 1α(HIF-1α),and vascular endothelial growth factor(VEGF)proteins.Results K

关 键 词：6-羟基染料木素 高原缺氧 网络药理学 HIF/VEGF信号通路 PI3K/AKT信号通路 

分 类 号：R594.3[医药卫生—内科学]