摘要
目的:探讨当归挥发油对缺氧/复氧(H/R)损伤大鼠心肌细胞H9C2自噬的调控作用。方法:以大鼠心肌细胞H9C2为对象,在CCK-8法筛选当归挥发油最佳给药浓度和给药时间的基础上,采用酶联免疫吸附测定法检测当归挥发油作用后细胞上清液中乳酸脱氢酶(LDH)的活性;以自噬抑制剂(3-甲基腺嘌呤,5 mmol/L)为阳性对照,采用MDC法和Western blotting法分别检测药物作用后细胞中MDC的平均荧光强度以及自噬相关蛋白[Beclin-1、微管相关蛋白轻链3Ⅱ(LC3Ⅱ)、LC3Ⅰ]的表达情况。结果:经0.6μmol/L当归挥发油作用6 h后,与空白组比较,H/R组细胞上清液中LDH活性和细胞中MDC平均荧光强度、Beclin-1表达水平、LC3Ⅱ/LC3Ⅰ比值均显著升高,细胞中p62表达水平显著降低(P<0.05或P<0.01);与H/R组比较,H/R+药物组细胞上清液中LDH活性,H/R+药物组和H/R+自噬抑制剂组细胞中MDC平均荧光强度、Beclin-1表达水平、LC3Ⅱ/LC3Ⅰ比值均显著降低,细胞中p62表达水平均显著升高(P<0.05或P<0.01)。结论:当归挥发油可通过调控自噬相关蛋白的表达来降低H/R损伤心肌细胞的自噬水平。
OBJECTIVE:To investigate the regulation effects of volatile oil from Angelica sinensis on autophagy of myocardial cell H9C2 in rats with hypoxia and reoxygenation(H/R)injury.METHODS:Using myocardial cell H9C2 as subject,CCK-8 method was used to screen the optimal concentration and administration time of volatile oil from A.sinensis.The acitivity of LDH in cell supernatant was determined after treated with volatile oil from A.sinensis by ELISA.Using autophagy inhibitors(3-methyladenine,5 mmol/L)as positive control,MDC method and Western blotting assay were used to detect average fluorescence intensity of MDC and the expression of autophagy related proteins[Beclin-1,microtubule associated protein light chain 3Ⅱ(LC3Ⅱ),LC3Ⅰ]in H9C2 cells after treated with medicines.RESULTS:After treated with 0.6μmol/L violate oil from A.sinensis for 6 h,compared with blank group,LDH activity in cell supernatant,average fluorescence intensity of MDC,the expression of Beclin-1,LC3Ⅱ/LC3Ⅰratio in cells were increased significantly in H/R group,while the expression of p62 was decreased significantly(P<0.05 or P<0.01).Compared with H/R group,the activity of LDH in cell supernatant of H/R+drug group as well as average fluorescence intensity of MDC,the expression of Beclin-1,LC3Ⅱ/LC3Ⅰratio in cells in H/R+drug group and H/R+autophagy inhibitor group were decreased significantly,while the expression of p62 were increased significantly(P<0.05 or P<0.01).CONCLUSIONS:The volatile oil from A.sinensis can reduce the autophagy level of H/R injury myocardial cells by regulating the expression of autophagy related proteins.
作者
浦延鹏
周佳明
PU Yanpeng;ZHOU Jiaming(Outpatient Department One of TCM,the First Affiliated Hospital of Jinzhou Medical University,Liaoning Jinzhou 121000,China;Dept.of Rehabilitation Medicine,Longwan District of Huludao Municipal Central Hospital,Liaoning Huludao 125003,China)
出处
《中国药房》
CAS
北大核心
2020年第20期2492-2497,共6页
China Pharmacy
基金
国家中医药管理局中医学术流派传承工作室——辽宁彭氏眼针学术流派传承工作室建设项目(No.LPGZS2012-09)
辽宁省科学技术计划项目(No.2016010306-301)。
关键词
当归挥发油
缺氧/复氧
大鼠心肌细胞H9C2
自噬
Volatile oil from Angelica sinensis
Hypoxia/reoxygenation
Myocardial cell H9C2 of rats
Autophagy
作者简介
浦延鹏,主治医师,博士研究生。研究方向:中西医结合临床神经内科学。E-mail:361364304@qq.com;通信作者:周佳明,主治医师,硕士。研究方向:中医内科学康复方向。E-mail:365021741@qq.com。
