高压氧对D-半乳糖诱导的衰老模型小鼠心肌纤维化及氧化应激相关指标表达的影响  

Effect of hyperbaric oxygen therapy on myocardial fibrosis and expression of oxidative stress-related indicators in D-galactose-induced senescent model mice

作　　者：郑晓月 王晓琳[1] 陈茗 王桂美[1] 安妮娜[1] 胡松[1] 龙颖 毛拥军[1] Zheng Xiaoyue;Wang Xiaolin;Chen Ming;Wang Guimei;An nina;Hu Song;Long Ying;Mao Yongjun(Department of Geriatric Medicine,the Affiliated Hospital of Qingdao University,Qingdao 266000,China;Department of Hyperbaric Oxygen,Shenzhen People's Hospital,Shenzhen 518020,China)

机构地区：[1]青岛大学附属医院老年医学科,青岛266000 [2]深圳市人民医院高压氧科,深圳518020

出　　处：《中华老年医学杂志》2022年第12期1512-1517,共6页Chinese Journal of Geriatrics

基　　金：国家自然科学基金资助项目(31571829)。

摘　　要：目的探讨高压氧疗法(HBOT)对D-半乳糖(D-gal)诱导的衰老小鼠心肌纤维化及氧化应激的保护作用及其机制。方法3月龄雄性昆明小鼠27只随机分为对照组、D-gal组和D-gal+HBOT组,对照组皮下注射灭菌生理盐水(5 ml·kg^(-1)·d^(-1))8周;其余2组皮下注射D-gal(200 mg·kg^(-1)·d^(-1))8周;D-gal+HBOT组在第7~8周进行HBOT。实验结束后取小鼠心脏组织通过苏木精-伊红(HE)染色观察病理学改变,马松染色和天狼星红染色观察纤维化改变;氧化应激试剂盒检测小鼠血清中过氧化氢酶(CAT)和总超氧化物歧化酶(T-SOD)的活性以及丙二醛(MDA)的含量;蛋白免疫印迹(western blot)法检测小鼠心脏组织中衰老相关蛋白p53和p16,心脏功能相关蛋白心房钠尿肽(ANP)和脑钠肽(BNP)以及氧化应激相关蛋白超氧化物歧化酶1(SOD1)、超氧化物歧化酶2(SOD2)和过氧化氢酶(CAT)蛋白表达水平。结果心脏形态学染色结果显示,与对照组比较,D-gal组小鼠表现出老化的特征,纤维化面积增加,HBOT干预后老化及纤维化较D-gal组明显改善。氧化应激试剂盒结果显示,与对照组比较,D-gal组小鼠血清中CAT和T-SOD活性明显降低、MDA含量明显增加,经HBOT处理后,D-gal+HBOT组血清中CAT和T-SOD活性较D-半乳糖组增加,MDA含量则降低(F=126.85、32.89、157.50,均P<0.05)。此外,与对照组比较,D-gal组心脏组织中p53、p16、ANP和BNP表达明显增加,CAT、SOD1和SOD2表达明显降低,HBOT干预后较D-gal组p53、p16、ANP和BNP表达降低,CAT、SOD1和SOD2表达增加(F=36.37、14.81、23.28、58.41、12.79、80.08、6.63,均P<0.05)。结论HBOT减轻了衰老模型小鼠的心脏损伤,可能与减弱心肌纤维化、诱导抗氧化酶表达、减少氧化应激有关。Objective To investigate the protective effect of hyperbaric oxygen therapy(HBOT)on myocardial fibrosis and oxidative stress induced by D-galactose(D-gal)in senescent model mice and its possible mechanism.Methods Three-month-old male Kunming mice(n=27)were randomized into control,D-gal,and D-gal+HBOT groups.The control group received subcutaneous sterilized saline(5 ml·kg^(-1)·d^(-1))for 8 weeks;the remaining 2 groups received subcutaneous D-gal(200 mg·kg^(-1)·d^(-1))for 8 weeks.The D-gal+HBOT group underwent HBOT intervention at week 7~8.At the end of the experiment,the histopathological changes were analyzed by hematoxylin-eosin(HE)staining,and the fibrosis changes were analyzed by Masson staining and Sirius red staining.Oxidative stress kit was used to detect catalase(CAT),total superoxide dismutase(T-SOD)activity and malon-di-aldehyde(MDA)content in serum of mice.Western blotting was used to detect the expression levels of the aging-related proteins p53 and p16 in mouse heart tissue,the heart-function-related proteins atrial natriuretic peptide(ANP)and brain natriuretic peptide(BNP),and the oxidative stress-related protein superoxide dismutase 1(SOD1),superoxide dismutase 2(SOD2)and catalase(CAT).Results Cardiac morphologic staining indicated that as compared with the control group,mice of D-gal group exhibited features of senescence and the increased fibrosis area,and senescence and fibrosis were obviously improved after HBOT intervention as compared with the D-gal group.The findings of the oxidative stress kit measurement indicated that as compared with the control group,the D-gal group had markedly decreased activities of CAT and T-SOD,significantly increased MDA content in the serum.After HBOT treatment,as compared with d-gal group,serum CAT and T-SOD activities were increased,while MDA content was decreased(F=126.85,32.89,157.50,all P<0.05).Furthermore,as compared with the control group,the D-gal group had obviously increased contents of p53,p16,ANP and BNP,while the content of CAT,SOD1 and SOD2 were

关 键 词：高压氧 心血管疾病 衰老 D-半乳糖 

分 类 号：R54[医药卫生—心血管疾病]