Page 66 - 2020年20期

当归挥发油对缺氧/复氧损伤大鼠心肌细胞 H9C2 自噬的调控作

用研究 Δ

浦延鹏 ，周佳明 （1.锦州医科大学附属第一医院中医科一门诊，辽宁 锦州 121000；2.葫芦岛市中心医院龙
2 #
1＊
湾院区康复医学科，辽宁 葫芦岛 125003）

中图分类号 R285.5 文献标志码 A 文章编号 1001-0408（2020）20-2492-06
DOI 10.6039/j.issn.1001-0408.2020.20.11
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0.6 μmol/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。
关键词 当归挥发油；缺氧/复氧；大鼠心肌细胞H9C2；自噬

Study on Regulation Effects of Volatile Oil from Angelica sinensis on Autophagy of Myocardial Cell H9C2
in Rats with Hypoxia/reoxygenation Injury
PU Yanpeng ，ZHOU Jiaming（1. Outpatient Department One of TCM，the First Affiliated Hospital of Jinzhou
2
1
Medical University，Liaoning Jinzhou 121000，China；2. Dept. of Rehabilitation Medicine，Longwan District of
Huludao Municipal Central Hospital，Liaoning Huludao 125003，China）

ABSTRACT 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 0.6 μmol/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/LC3Ⅰ ratio 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.
KEYWORDS Volatile oil from Angelica sinensis；Hypoxia/reoxygenation；Myocardial cell H9C2 of rats；Autophagy

缺血性心脏病大多是由冠状动脉供血不足所致，可 引起心肌损伤。如果能及时恢复心脏供血，可恢复心脏
功能；但也可能导致心肌损伤加重，造成缺血/再灌注
Δ 基金项目：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 （I/R）损伤 [1-2] 。在很多情况下，机体会出现 I/R 损伤，如
省科学技术计划项目（No.2016010306-301） 创伤、收缩后血管回流、经冠状动脉成形术、溶栓治疗
＊主治医师，博士研究生。研究方向：中西医结合临床神经内科
等，而当受损的缺血心肌组织经过干预再次恢复血液灌
学。E-mail：361364304@qq.com
注时，往往会伴随着组织损伤加重的可能，引起心功能
# 通信作者：主治医师，硕士。研究方向：中医内科学康复方向。
E-mail：365021741@qq.com 不全、再灌注心律失常和心肌梗死加重等 [3-4] 。因此，如

·2492 · China Pharmacy 2020 Vol. 31 No. 20 中国药房 2020年第31卷第20期