花椒提取物羟基-α-山椒素通过调节Nrf2/HO-1通路减轻心肌损伤  

Improvement of Myocardial Injury by Hydroxy-α-Sanshool from Extracts of ZANTHOXYLI PERICARPIUM via Regulating Nrf2/HO-1 Pathway

作　　者：陈文文 汤丹丹 张青 华平 蔡健 吴纯洁[2] CHEN Wenwen;TANG Dandan;ZHANG Qing;HUA Ping;CAI Jian;WU Chunjie(Department of Pharmacy,Chengdu Women's and Children's Central Hospital,School of Medicine,University of Electronic Science and Technology of China,Chengdu 611137;School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137;Department of Pathology,Chengdu Women's and Children's Central Hospital,School of Medicine,University of Electronic Science and Technology of China,Chengdu 611137)

机构地区：[1]电子科技大学医学院附属妇女儿童医院·成都市妇女儿童中心医院药学部,成都611731 [2]成都中医药大学药学院,成都都611137 [3]电子科技大学医学院附属妇女儿童医院·成都市妇女儿童中心医院病理科,成都611731

出　　处：《中药药理与临床》2024年第7期48-56,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川省卫生健康委员会医学科技项目(编号:21PJ130)。

摘　　要：目的:研究花椒提取物羟基-α-山椒素对异丙肾上腺素所致心肌缺血大鼠和过氧化氢(H_(2)O_(2))诱导的大鼠心肌细胞(H9c2)氧化损伤的保护作用及可能机制。方法:36只SD雄性大鼠随机分为正常对照组、模型对照组、盐酸地尔硫卓20 mg/kg、羟基-α-山椒素5、10和20 mg/kg组。各组大鼠连续给药14 d,在13、14 d腹腔注射异丙肾上腺素(ISO)40 mg/kg造成心肌缺血模型,超声心动图检查评估各组大鼠心功能;试剂盒检测血清丙二醛(MDA)、还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、心肌肌钙蛋白Ⅰ(cTnI)、乳酸脱氢酶(LDH)、肌酸激酶同工酶(CK-MB)、肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和IL-6含量;实时荧光定量qRT-PCR法检测心肌组织Tnfa、Il1和Il6 mRNA表达;苏木素-伊红(HE)和马松(Masson)染色观察心肌组织病理改变。体外培养H9c2细胞,用H_(2)O_(2)600μmol/L处理H9c2细胞6 h构建氧化损伤体外细胞模型,随机分为正常对照组、模型对照组、羟基-α-山椒素2.5、5、10μg/mL组。采用细胞计数试剂盒(CCK-8)法筛选羟基-α-山椒素最佳给药浓度;采用流式细胞仪和激光共聚焦显微镜观测细胞凋亡情况;流式细胞仪检测细胞活性氧(ROS)浓度;ELISA法检测MDA、谷胱甘肽过氧化物酶(GSH-Px)和SOD含量或活力;蛋白质印迹法检测核因子E2相关因子2(Nrf2)、B淋巴细胞瘤-2基因相关X蛋白(BAX)、血红素氧合酶1(HO-1)、切割胱天蛋白酶3(Cleaved-caspase 3)和B淋巴细胞瘤-2基因(BCL-2)蛋白表达水平。结果:与正常对照组相比,模型对照组大鼠左心室短轴缩短率(LVFS)和射血分数(LVEF)显著降低(P<0.01),血清MDA、cTnl、LDH、CK-MB含量或活力显著升高(P<0.01),GSH、SOD活力显著降低(P<0.01),心肌组织中Il1b、Il6及TnfamRNA表达明显上调(P<0.05);与模型对照组相比,羟基-α-山椒素各剂量组LVFS和LVEF明显升高(P<0.05),羟基-α-山椒素20 mg/kg组MDA、cTnl、LDH、CK-MB含量或活力明显Objective:To investigate the protective effect and mechanism of hydroxy-α-sanshool( HAS) from extracts of ZANTHOXYLI PERICARPIUM against rats with isoprenaline(ISO)-induced myocardial ischemia(MI) and oxidative damage in myocardial cells of rats(H9c2)induced by hydrogen peroxide(H_(2)O_(2)).Methods:A total of 36 SD male rats were randomly divided into a normal control group,a model control group,diltiazem hydrochloride group of 20 mg/kg,and HAS groups of 5,10,and 20 mg/kg. Rats were administered for 14 days,and ISO of 40 mg/kg was injected intraperitoneally on the 13th and 14th day to induce the MI model. Echocardiography was used to evaluate the cardiac function of rats in each group. The malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD),cardiac troponin I(cTnI),lactate dehydrogenase(LDH),and creatine kinase isoenzyme(CK-MB) were detected by kits. The contents of tumor necrosis factor-α(Tnf-α),interleukin-1β(Il-1β),andIl-6 in the myocardial tissue were detected by real-time fluorescence quantitative qRT-PCR. Hematoxylin-Eosin(HE) staining and Masson staining were used to observe the pathological changes in the myocardial tissue. H9c2 was culturedin vitro,and oxidative damage was induced by treating H9c2 with H_(2)O_(2)of 600 μmol/L for six hours to establish anin vitrocell model. The cells were randomly divided into a normal control group,model control group,and HAS groups of 2.5,5,and 10 μg/mL. The optimal drug concentration of HAS was selected using the cell counting kit(CCK-8). Cell apoptosis was observed by using flow cytometry and laser confocal microscopy. The reactive oxygen species(ROS) concentration of the cells was measured using flow cytometry. The contents or activities of antioxidant enzymes MDA,GSH-Px,and SOD were determined using the enzyme-linked immunosorbent assay(ELISA). The expression levels of Nrf2,Bax,HO-1,cleaved-Caspase-3,and Bcl-2 were detected by Western blot(WB).Results:Compared with those in the normal control group,the left ventricular fraction shortening(LVFS) a

关 键 词：羟基-α-山椒素 心肌缺血 炎症因子 氧化应激 核因子E2相关因子2 

分 类 号：R285.5[医药卫生—中药学]