摘要
目的:考察茅苍术挥发油对缺氧/复氧(H/R)损伤H9c2心肌细胞的抗氧化与抗凋亡活性,并比较生品和麸炒品的作用差异。方法:以连二亚硫酸钠(Na_(2)S_(2)O_(4))诱导H9c2缺氧/复氧损伤模型,噻唑蓝(MTT)法检测细胞活力,4′,6-二脒基-2-苯基吲哚(DAPI)染色评估细胞凋亡,酶联免疫吸附测定(Elisa)法检测细胞培养液中超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)和丙二醛(MDA)的含量;实时荧光定量聚合酶链式反应(RT-PCR)法检测细胞中半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)和Bcl-2同源的水溶性相关蛋白(Bax)mRNA的表达;蛋白免疫印迹法(Western blot)检测细胞中裂解的Caspase-3(cleaved Caspase-3)、BAX、BCL-2的蛋白表达。结果:与正常对照组比较,模型对照组心肌细胞的存活率显著降低,SOD活力、Bcl-2 mRNA、蛋白表达明显降低,LDH、MDA含量、Caspase-3 mRNA、Bax mRNA、cleaved Caspase-3、BAX蛋白表达明显升高(P<0.05)。生品及麸炒茅苍术挥发油1μg/mL~100μg/mL能够提高H9c2心肌细胞的存活率;提高SOD的活力,降低LDH和MDA的含量;上调Bcl-2 mRNA和蛋白的表达,下调Caspase-3、Bax mRNA的表达;下调cleaved Caspase-3、BAX的蛋白表达,与同剂量的生品挥发油比较,麸炒品挥发油具有更好的作用。结论:茅苍术挥发油对H/R损伤心肌细胞具有抗氧化和抗凋亡的作用,麸炒品挥发油的作用优于生品挥发油。
Objective:To investigate the anti-oxidative and anti-apoptotic effects of essential oil from Maocangzhu on H9c2 cardiomyocytes injured by hypoxia/reoxygenation(H/R),and compare the therapeutic effects of raw Maocangzhu and bran-processed Maocangzhu.Methods:Sodium dithionite(Na_(2)S_(2)O_(4))was used to induce the H/R injury model in H9c2 cardiomyocytes.The cell viability was detected by the MTT assay,and 4′,6-diamidino-2-phenylindole(DAPI)staining was used to evaluate cell apoptosis.The levels of superoxide dismutase(SOD),lactic dehydrogenase(LDH),and malondialdehyde(MDA)were determined by the enzyme-linked immunosorbent assay(ELISA).The mRNA and protein expression of cysteine-aspartic acid protease 3(Caspase-3),B-cell lymphoma 2(Bcl-2),and Bcl-2-associated X protein(Bax)was detected by RT-PCR and Western blot,respectively.Results:Compared with the normal control group,the model group showed blunted cell viability of cardiomyocytes,decreased SOD activity,and Bcl-2 mRNA and protein expression,increased content of LDH and MDA,and up regulated expression of Caspase-3 mRNA,Bax mRNA,cleaved Caspase-3,and Bax protein(P<0.05).Both essential oils from raw Maocangzhu and bran processed Maocangzhu(1μg/mL~100μg/mL)could potentiate the cell viability of cardiomyocytes and SOD activity,decrease the content of LDH and MDA,up regulate the mRNA and protein expression of Bcl-2,down-regulate the mRNA expression of Caspase-3 and Bax,and reduce the protein expression of cleaved Caspase-3 and Bax.The essential oil from bran-processed Maocangzhu was superior to that from raw Maocangzhu at the same dose in efficacy.Conclusion:The essential oil from Maocangzhu has anti-oxidative and anti-apoptotic effects on cardiomyocytes injured by H/R,and the essential oil from bran-processed Maocangzhu shows a better effect than that from raw Maocangzhu.
作者
于艳
贾天柱
魏新智
鞠业涛
韩宇
于永铎
Yu Yan;Jia Tianzhu;Wei Xinzhi;Ju Yetao;Han Yu;Yu Yongduo(Liaoning University of Traditional Chinese Medicine,Shenyang 110847;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110847)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第1期124-130,共7页
Pharmacology and Clinics of Chinese Materia Medica
基金
辽宁省博士启动基金(编号:20180540125)
国家自然科学基金项目(编号:81803727)
辽宁省教育厅一般项目(编号:L201906)
关键词
茅苍术
炮制
缺血再灌注
缺氧复氧损伤
氧化应激
凋亡
Maocangzhu
processed
ischemia reperfusion
hypoxia/reoxygenation
oxidative stress
apoptosis
作者简介
通信作者:于永铎,博士,教授、主任医师,博士研究生导师,研究方向:中医药防治肛肠疾病,E-mail:yuyongduo@163.com;于艳,博士,主任中药师,硕士研究生导师,研究方向:中药炮制与中药分析,E-mail:95142012@qq.com。
