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基于AMPK/SIRT1/PGC-1α信号通路研究香青兰总黄酮对大鼠心
肌缺血再灌注损伤的保护机制 Δ
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赵云丽 1,2* ,袁 勇 ,马晓莉 ,黄川生 ,文志萍 ,郭新红 ,王新春 (1.石河子大学药学院,新疆 石河子
832002;2.石河子大学医学院第一附属医院药剂科,新疆 石河子 832008)
中图分类号 R285 文献标志码 A 文章编号 1001-0408(2021)03-0278-06
DOI 10.6039/j.issn.1001-0408.2021.03.05
摘 要 目的:研究香青兰总黄酮(TFDM)对腺苷酸活化蛋白激酶(AMPK)/沉默信息调节因子1(SIRT1)/过氧化物酶体增殖物激
活受体γ辅激活因子1α(PGC-1α)信号通路的影响,探究其保护大鼠心肌缺血再灌注损伤(MIRI)的作用机制。方法:将50只健康
雄性 SD 大鼠随机分为假手术组、模型组、TFDM 组[60 mg/(kg·d),以提取物计]、Compound C+TFDM 组[灌胃 60 mg/(kg·d)
TFDM+再灌注前15 min尾静脉注射250 μg/kg Compound C(AMPK抑制剂)]、EX-527+TFDM组[灌胃60 mg/(kg·d)TFDM+再灌
注前20 min腹腔注射5 mg/kg EX-527(SIRT1抑制剂)],每组10只。每天灌胃给药1次,连续7 d。末次灌胃给药后,假手术组大鼠
行假手术,其余4组大鼠均采用结扎冠状动脉左前降支缺血30 min、再灌注2 h构建MIRI模型。再灌注结束后,采用苏木精-伊红
染色法观察大鼠心肌组织病理学变化;采用反相高效液相色谱法测定其心肌组织中腺苷三磷酸(ATP)、腺苷二磷酸(ADP)、腺苷
+
一磷酸(AMP)及烟酰胺腺嘌呤二核苷酸(NAD )的含量;采用实时荧光定量-聚合酶链式反应法检测大鼠心肌组织中 AMPK、
SIRT 1和PGC-1α mRNA表达水平;采用Western blotting法检测大鼠心肌组织中AMPK蛋白的磷酸化水平和SIRT 1、PGC-1α蛋白
表达水平。结果:与假手术组比较,模型组大鼠心肌纤维排列紊乱、横向条纹消失,细胞肿胀破裂、坏死,细胞核变形移位;心肌组
织中 ATP、NAD 含量和 AMPK、SIRT1、PGC-1α mRNA 表达水平以及 SIRT1、PGC-1α蛋白表达水平均显著降低(P<0.05 或 P<
+
0.01),ADP、AMP含量及AMPK蛋白的磷酸化水平均显著升高(P<0.01)。与模型组比较,TFDM组大鼠心肌病理学形态明显改
+
善;心肌组织中ATP、NAD 含量和AMPK、SIRT1、PGC-1α mRNA表达水平以及AMPK蛋白的磷酸化水平和SIRT1、PGC-1α蛋白
表达水平均显著升高(P<0.05 或 P<0.01),ADP、AMP 含量均显著降低(P<0.01)。与 TFDM 组比较,Compound C+TFDM 组和
EX-527+TFDM组大鼠上述指标的改善作用均被逆转(P<0.05或P<0.01)。结论:TFDM可能是通过激活AMPK/SIRT1/PGC-1α
信号通路,调节能量代谢,从而发挥其对心肌的保护作用。
关键词 香青兰总黄酮;能量代谢;腺苷酸活化蛋白激酶;沉默信息调节因子 1;过氧化物酶体增殖物激活受体γ辅激活因子 1α;
机制
Study on Protective Mechanism of Dracocephalum moldavica Total Flavonoids against Myocardial
Ischemia-reperfusion Injury in Rats Based on AMPK/SIRT1/PGC-1α Signaling Pathway
ZHAO Yunli ,YUAN Yong ,MA Xiaoli ,HUANG Chuansheng ,WEN Zhiping ,GUO Xinhong ,WANG
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Xinchun(1.College of Pharmacy,Shihezi University,Xinjiang Shihezi 832002,China;2.Dept. of Pharmacy,
the First Affiliated Hospital of the Medical College,Shihezi University,Xinjiang Shihezi 832008,China)
ABSTRACT OBJECTIVE:To study the effects of Dracocephalum moldavica total flavonoids(TFDM)on AMPK/SIRT1/PGC-1α
signaling pathway,and to explore the mechanism of its protective effect on myocardial ischemia reperfusion injury(MIRI)rats.
METHODS:Totally 50 healthy male SD rats were randomly divided into sham operation group,model group,TFDM group [60
mg/(kg·d),by extract],Compound C+TFDM group [ig administration of 60 mg/(kg·d)TFDM+intravenous injection of 250
μg/kg Compound C(AMPK inhibitor)via tail vein 15 min before reperfusion],EX-527+TFDM group [ig administration of 60 mg/
(kg·d)TFDM+ip injection of 5 mg/kg EX-527(SIRT1 inhibitor)20 min before reperfusion],with 10 rats in each group. They
were given relevant medicine intragastrically,once a day,for consecutive 7 days. After last ig administration,sham operation
group underwent sham operation,other 4 groups were established MIRI model by ligating left anterior descending coronary artery,
ischemia for 30 min and reperfusion for 2 h. After reperfusion,the myocardial histopathological changes were observed by HE
staining;RP-HPLC method was used to determine the contents of ATP,ADP,AMP and NAD + in cardiac tissue. mRNA
expressions of AMPK,SIRT1 and PGC-1α were detected by
Δ 基 金 项 目 :国 家 自 然 科 学 基 金 资 助 项 目(No.81860747,
quantitative real-time PCR assay. Western blotting assay was
No.81960766)
used to detect the phosphorylation level of AMPK protein and
*硕士研究生。研究方向:中药民族药新药研究与开发。E-mail:
1543279962@qq.com the expressions of SIRT 1 and PGC-1α protein in myocardium.
# 通信作者:主任药师,硕士生导师,博士。研究方向:中药民族 RESULTS: Compared with sham operation group, model
药新药研究与开发。E-mail:841175436@qq.com group showed myocardial fibers arranged disorder and
·278 · China Pharmacy 2021 Vol. 32 No. 3 中国药房 2021年第32卷第3期