冠心丹参制剂通过miR-330-3p/ALDH2保护大鼠心脏免受缺血/再灌注损伤

袁芳; 杨艳华; 赵鑫扬; 李磊

doi:10.16016/j.1000-5404.202101061

您当前的位置：

首页 >

文章列表页 >

冠心丹参制剂通过miR-330-3p/ALDH2保护大鼠心脏免受缺血/再灌注损伤

基础医学 | 更新时间：2026-01-09

- 冠心丹参制剂通过miR-330-3p/ALDH2保护大鼠心脏免受缺血/再灌注损伤
- Guanxin Salvia Miltiorrhiza Preparation protects rat cardiomyocytes against ischemia/reperfusion injury through miR-330-3p/ALDH2
- 第三军医大学学报 2021年43卷第13期页码：1243-1252
- 作者机构：
  
  430000 武汉，武汉市第四医院，华中科技大学同济医学院附属普爱医院心内科
- 作者简介：
  
  [李磊，E-mail 1517836083@qq.comLI Lei, E-mail: 1517836083@qq.com
- 基金信息：
  
  武汉市卫生计生委项目(WZ18Q10)
- DOI：10.16016/j.1000-5404.202101061
  中图分类号： R285.5;R289.5;R541.5
- 收稿：2021-01-07，
  
  修回：2021-03-16，
  
  纸质出版：2021-07-15
- 稿件说明：
移动端阅览
袁芳, 杨艳华, 赵鑫扬, 等. 冠心丹参制剂通过miR-330-3p/ALDH2保护大鼠心脏免受缺血/再灌注损伤[J]. 第三军医大学学报, 2021,43(13):1243-1252.

Fang YUAN, Yanhua YANG, Xinyang ZHAO, et al. Guanxin Salvia Miltiorrhiza Preparation protects rat cardiomyocytes against ischemia/reperfusion injury through miR-330-3p/ALDH2[J]. Journal of Third Military Medical University, 2021, 43(13): 1243-1252.
袁芳, 杨艳华, 赵鑫扬, 等. 冠心丹参制剂通过miR-330-3p/ALDH2保护大鼠心脏免受缺血/再灌注损伤[J]. 第三军医大学学报, 2021,43(13):1243-1252. DOI： 10.16016/j.1000-5404.202101061.

Fang YUAN, Yanhua YANG, Xinyang ZHAO, et al. Guanxin Salvia Miltiorrhiza Preparation protects rat cardiomyocytes against ischemia/reperfusion injury through miR-330-3p/ALDH2[J]. Journal of Third Military Medical University, 2021, 43(13): 1243-1252. DOI： 10.16016/j.1000-5404.202101061.

摘要

目的

探讨冠心丹参(Guanxin Salvia miltiorrhiza

GXDS)的心脏保护作用与肥大细胞脱粒抑制是否有关及其机制。

方法

使用miR-330-3p mimics、ALDH2 Vector转染或者肥大细胞促分泌素化合物48/80(C48/80)处理心肌细胞，采用PCR和Western blot检测miR-330-3p和ALDH2的表达，MTT和流式细胞仪检测细胞增殖和凋亡，ELISA检测肥大细胞脱粒和类胰蛋白酶释放，双荧光素酶报告基因检测两者的靶向结合。通过结扎大鼠左前降支冠状动脉30 min并再灌注120 min建立缺血再灌注(I/R)损伤模型，使用肥大细胞促分泌素化合物48/80(C48/80)、GXDS治疗，通过免疫组化检测不同处理组小鼠心肌组织损伤。

结果

C48/80抑制心肌细胞H9C2(2-1)和肥大细胞RBL-2H3的活力并促进其凋亡(

0.05)，增加cTnI和类胰蛋白酶的释放(

0.05)，而GXDS处理可通过miR-330-3p/ALDH2促进心肌细胞的活力并抑制其凋亡，同时抑制肥大细胞脱粒(

0.05)；进一步的体内实验结果显示: 与假手术组相比，I/R组血流动力学异常，心律不齐，心脏水肿，梗死面积、组织病理学损伤以及cTnI和miR-330-3p的水平均显著升高(

0.05)，ALDH2表达水平下调(

0.05)，而GXDS预处理可以通过抑制肥大细胞脱粒和类胰蛋白酶释放发挥心肌保护作用。

结论

GXDS通过miR-330-3p/ALDH2抑制大鼠心脏肥大细胞脱粒，从而改善缺血/再灌注引起的损伤。

Abstract

Objective

To explore the cardioprotective effectiveness of Guanxin

Salvia miltiorrhiza

(GXDS) Preparation (a prescribed Chinese herbal formula) and its relationship with inhibition of mast cell degranulation

and investigate the underlying mechanism.

Methods

Cardiomyocytes H9C2 (2-1) cocultured with mast RBL-2H3 cells cells were transfected with miR-330-3p mimics or ALDH2 Vector

or treated with GXDS or Compound 48/80

a mast cell degranulator. The expression of miR-330-3p and ALDH2 was detected by PCR and Western blotting

and cell proliferation and apoptosis were detected by MTT assay and flow cytometry. Mast cell degranulation and trypsin release were detected by ELISA

and the targeted binding of mast cell degranulation and trypsin release was detected by double luciferase reporter gene. Then

a rat model of myocardial ischemia/reperfusion injury was established by ligating the left anterior descending coronary artery for 30 min followed by reperfusion for 120 min. Then Compound 48/80 or GXDS was used to treat the rats with myocardial injury or sham operation. The myocardial injury was detected by immunohistochemistry in different treatment groups.

Results

Compound 48/80 inhibited the viability and promoted the apoptosis of H9C2 (2-1) and RBL-2H3 cells

and increased the releases of cTnI and trypsin (

0.05). GXDS treatment could promote the viability and inhibit the apoptosis of cardiomyocytes through miR-330-3p/ALDH2 (

0.05)

and inhibit mast cell degranulation (

0.05). Compared with the sham operation rats

the hemodynamic abnormalities

arrhythmia

cardiac edema

infarct size

histopathological damages and levels of cTnI and miR-330-3p were significantly increased and the expression of ALDH2 was down-regulated in the rats of the model group (

0.05). GXDS pretreatment protected myocardium by inhibiting mast cell degranulation and trypsin release.

Conclusion

GXDS inhibits the degranulation of rat cardiac mast cells through miR-330-3p/ALDH2

and thus improves the myocardial ischemia/reperfusion injury.

关键词

Keywords

references

刘晓瑜, 顾民华, 徐煜凌, 等. 定心方Ⅰ号方通过调控GSK3β/Nrf2/HO-1通路防治大鼠心肌缺血再灌注损伤的机制研究[J]. 中药新药与临床药理, 2020, 31(11): 1265-1270. DOI:10.19378/j.issn.1003-9783.2020.11.001.

LIU X Y, GU M H, XU Y L, et al . Dingxin recipe Ⅰ prevents myocardial ischemia reperfusion injury through GSK3β/Nrf2/HO-1 pathway[J ] . Tradit Chin Drug Res Clin Pharmacol, 2020, 31(11): 1265-1270. DOI:10.19378/j.issn.1003-9783.2020.11.001.

蒋佳玲, 李俊, 许鸣, 等. 广枣叶总黄酮对心肌缺血再灌注损伤模型大鼠的保护作用及其作用机制[J]. 沈阳药科大学学报, 2020, 37(11): 1009-1014. DOI:10.14066/j.cnki.cn21-1349/r.2020.11.008.

JIANG J L, LI J, XU M, et al . Myocardial protective function and mechanism of total flavonoids from Jujube leaves after myocardial ischemia-reperfusion injury in rats[J ] . J Shenyang Pharm Univ, 2020, 37(11): 1009-1014. DOI:10.14066/j.cnki.cn21-1349/r.202 0.11.008.

SONG S H, LIU F, ZHAO Y Y, et al . Bruton's tyrosine kinase inhibitor attenuates warm hepatic ischemia/reperfusion injury via modulation of the NLR family pyrin domain containing 3 inflammasome[J ] . Transplant Proc, 2020, 52(10): 2947-2954. DOI:10.1016/j.transproceed.2019.10.024..

王励, 谌小龙, 刘慧玲, 等. β-arrestin-2通过上调PI3K/Akt信号抑制自噬以减轻小鼠肝脏缺血-再灌注损伤[J]. 器官移植, 2020, 11(6): 692-697. DOI:10.3969/j.issn.1674-7445.2020.06.007.

WANG L, CHEN X L, LIU H L, et al . Β-arrestin-2 inhibits autophagy by up-regulating PI3K/Akt signal to alleviate liver ischemia-reperfusion injury in mice[J ] . Organ Transplant, 2020, 11(6): 692-697. DOI:10.3969/j.issn.1674-7445.2020.06.007.

夏万松, 夏英, 韦四喜, 等. ALDH2基因rs671单核苷酸多态性位点发生(A→G)RNA编辑[J]. 国际检验医学杂志, 2020, 41(22): 2693-2696, 2700. DOI:10.3969/j.issn.1673-4130.2020.22.002.

XIA W S, XIA Y, WEI S X, et al . ALDH2 gene rs671 single nucleotide polymorphism site occurs RNA editing (A to G)[J ] . Int J Lab Med, 2020, 41(22): 2693-2696, 2700. DOI:10.3969/j.issn.1673-4130.2020.22.002.

ZHOU L, ISHIZAKI H, SPITZER M, et al . ALDH2 mediates 5-nitrofuran activity in multiple species[J ] . Chem Biol, 2012, 19(7): 883-892. DOI:10.1016/j.chembiol.2012.05.017.

宋海英, 王培利, 刘剑刚, 等. 冠心丹参滴丸对缺血心肌AMPK-解偶联蛋白2的调控作用[J]. 中西医结合心脑血管病杂志, 2019, 17(15): 2272-2276.

SONG H Y, WANG P L, LIU J G, et al . The modulation of AMPK-uncoupling protein 2 in ischemia myocardium by Guanxindanshen dropping pills[J ] . Chin J Integr Med Cardio-Cerebrovasc Dis, 2019, 17(15): 2272-2276.

汤世民, 邢小燕, 邓雪红, 等. 冠心丹参方及其有效成分治疗冠心病的研究进展[J]. 中国中药杂志, 2016, 41(20): 3721-3726.

TANG S M, XING X Y, DENG X H, et al . Research progress of Guanxin Danshen formula and its effective components in treating coronary artery heart disease[J ] . China J Chin Mater Med, 2016, 41(20): 3721-3726.

CHENG Y, ZHU H, GAO W. MicroRNA-330-3p represses the proliferation and invasion of laryngeal squamous cell carcinoma through downregulation of Tra2β-mediated Akt signaling[J]. Mol CellProbes, 2020, 52: 101574. DOI:10.1016/j.mcp.2020.101574.

苟小江, 王培利, 王承龙. 冠心丹参滴丸治疗冠心病的系统评价与Meta分析[J]. 中药新药与临床药理, 2019, 30(10): 1262-1268. DOI:10.19378/j.issn.1003-9783.2019.10.018.

GOU X J, WANG P L, WANG C L. Systematic review and meta analysis of Guanxin Danshen dropping pills in the treatment of coronary heart disease[J]. New Drugs Clin Pharmacol Traditional Chin Med, 2019, 30(10): 1262-1268.

SONG K, LI L, QUAN Q, et al . Inhibited histone deacetylase 3 ameliorates myocardial ischemia-reperfusion injury in a rat model by elevating microRNA-19a-3p and reducing cyclin-dependent kinase 2[J ] . IUBMB Life, 2020, 72(12): 2696-2709. DOI:10.1002/iub.2402.

MIYAKE H, KOIKE Y, SEO S, et al . The effect of pre-and post-remote ischemic conditioning reduces the injury associated with intestinal ischemia/reperfusion[J ] . Pediatr Surg Int, 2020, 36(12): 1437-1442. DOI:10.1007/s00383-020-04762-5.

何贵新, 肖婷, 秦伟彬, 等. 基于PI3K/AKT信号通路探讨芪参益气滴丸改善心肌缺血再灌注损伤的实验研究[J]. 北京中医药大学学报, 2020, 43(9): 762-768. DOI:10.3969/j.issn.1006-2157.2020.09.010.

HE G X, XIAO T, QIN W B, et al . Experimental study on the effect of Qishen Yiqi Dripping Pill on myocardial ischemia-reperfusion injury via PI3K/AKT signaling pathway[J ] . J Beijing Univ Tradit Chin Med, 2020, 43(9): 762-768. DOI:10.3969/j.issn.1006-2157.2020.09.010..

WANG H L, LIU F L, LI R Q, et al . Electroacupuncture improves learning and memory functions in a rat cerebral ischemia/reperfusion injury model through PI3K/Akt signaling pathway activation[J ] . Neural Regen Res, 2021, 16(6): 1011-1016. DOI:10.4103/1673-5374.300454.

郭银雪, 葛平玉, 詹继红. 肾茶黄酮预防急性肾缺血模型大鼠再灌注损伤作用及其机制[J]. 中国老年学杂志, 2020, 40(20): 4399-4402.

GUO Y X, GE P Y, ZHAN J H. Preventive effect of renal tea flavonoids on reperfusion injury in rats with acute renal ischemia and its mechanism[J]. Chin J Gerontol, 2020, 40(20): 4399-4402.

彭兴, 林玲, 周祥群, 等. miR-133b靶向YES1抑制心肌缺血再灌注引起的心肌细胞凋亡和活性氧簇的积累[J]. 南方医科大学学报, 2020, 40(10): 1390-1398. DOI:10.12122/j.issn.1673-4254.2020.10.03.

PENG X, LIN L, ZHOU X Q, et al . miR-133b inhibits myocardial ischemia-reperfusion-induced cardiomyocyte apoptosis and accumulation of reactive oxygen species in rats by targeting YES1[J ] . J South Med Univ, 2020, 40(10): 1390-1398. DOI:10.12122/j.issn.1673-4254.2020.10.03.

WEI C H, WU G, CAI Q, et al . Retraction Note to: MicroRNA-330-3p promotes cell invasion and metastasis in non-small cell lung cancer through GRIA3 by activating MAPK/ERK signaling pathway[J ] . J Hematol Oncol, 2020, 13(1): 142. DOI:10.1186/s13045-020-00969-0.

SHAO Z Q, DOU S S, ZHU J G, et al . Apelin-13 inhibits apoptosis and excessive autophagy in cerebr al ischemia/reperfusion injury[J ] . Neural Regen Res, 2021, 16(6): 1044-1051. DOI:10.4103/1673-5374.300725.

王晓丽, 张薇, 张雪侠, 等. 生姜挥发油对脑缺血再灌注大鼠溶酶体-线粒体途径凋亡的影响[J]. 中药新药与临床药理, 2020, 31(10): 1153-1157. DOI:10.19378/j.issn.1003-9783.2020.10.004.

WANG X L, ZHANG W,ZHANG X X, et al . Effect of ginger volatile oil on lysosomal-mitochondrial apoptosis in rats with cerebral ischemia-reperfusion[J ] . Tradit Chin Drug Res Clin Pharmacol, 2020, 31(10): 1153-1157. DOI:10.19378/j.issn.1003-9783.2020.10.004.

何望安, 王茹, 沈波, 等. CCR5抗体对大鼠心肌缺血再灌注损伤保护作用的机制研究[J]. 华中科技大学学报(医学版), 2020, 49(4): 419-423, 454.

HE W A, WANG R, SHEN B, et al . Protective effects of CCR5 antibody on myocardial ischemia-reperfusion injury in rats[J ] . Acta Med Univ Sci et Technol Huazhong, 2020, 49(4): 419-423, 454.

龚慧琴, 周支香, 刘录山, 等. 淫羊藿次苷Ⅱ通过沉默信息调控子1-叉头蛋白O1通路减轻心肌缺血再灌注损伤[J]. 中华高血压杂志, 2020, 28(8): 744-749. DOI:10.16439/j.cnki.1673-7245.2020.08.015.

GONG H Q, ZHOU Z X, LIU L S, et al . Icariside Ⅱ reduces myocardial ischemia-reperfusion injury through activating silent information regulator 1/acetylated-forkhead box O1 pathway[J ] . Chin J Hypertens, 2020, 28(8): 744-749. DOI:10.16439/j.cnki.1673-7245.2020.08.015.

WEI C H, ZHANG R G, CAI Q, et al . MicroRNA-330-3p promotes brain metastasis and epithelial-mesenchymal transition via GRIA3 in non-small cell lung cancer[J ] . Aging, 2019, 11(17): 6734-6761. DOI:10.18632/aging.102201..

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

不同致痒剂诱导建立大鼠瘙痒模型的效果分析

SELEX技术筛选钙通道特异性核酸适配体及其抗肥大细胞活化的作用

缓激常泰拮抗细菌潜生体对肥大细胞激活作用的实验研究