丹参多酚酸对心房颤动大鼠心肌组织中VCAM-1和ICAM-1水平及ERK1/2-NF-κB信号通路的影响

doi:10.13481/j.1671-587x.20200517

摘要/Abstract

摘要： 目的：研究丹参多酚酸对心房颤动（房颤）大鼠心肌组织中血管细胞黏附分子1（VCAM-1）和细胞间黏附分子1（ICAM-1）水平及细胞外信号调节酶1/2-核转录因子-κB（ERK1/2-NF-κB）信号通路的影响，探讨丹参多酚酸防治房颤的可能机制。方法：48只SD大鼠随机分为对照组、房颤组和丹参多酚酸组，每组16只。舌下静脉注射氯化钙-乙酰胆碱混合液建立房颤大鼠模型。丹参多酚酸组大鼠用丹参多酚酸（4 mg·kg^-1·d^-1）灌胃，对照组和房颤组大鼠用等量生理盐水灌胃，每天1次，共4周。采用苏木精-伊红（HE）染色观察大鼠心肌组织病理形态表现，Masson染色观察心肌纤维化情况，免疫组织化学染色检测各组大鼠心肌组织中基质金属蛋白酶2（MMP-2）和基质金属蛋白酶9（MMP-9）蛋白表达水平，Western blotting法检测各组大鼠心肌组织中VCAM-1、ICAM-1、ERK1/2、磷酸化细胞外调节蛋白激酶1/2（p-ERK1/2）、NF-κB和磷酸化核转录因子κB（p-NF-κB）蛋白表达水平。结果：HE染色，对照组大鼠心肌细胞未见明显异常；房颤组大鼠心肌组织间质增多，可见炎性细胞浸润；与房颤组比较，丹参多酚酸组大鼠心肌组织间质稍多，炎性细胞浸润不明显。Masson染色，对照组大鼠心肌间质胶原纤维正常；房颤组大鼠心肌间质胶原纤维明显增多；与房颤组比较，丹参多酚酸组大鼠心肌间质胶原纤维较房颤组明显减少。与对照组比较，房颤组和丹参多酚酸组大鼠心肌组织中MMP-2、MMP-9、VCAM-1、ICAM-1、p-ERK1/2和p-NF-κB蛋白表达水平升高（P<0.05）；与房颤组比较，丹参多酚酸组大鼠心肌组织中MMP-2、MMP-9、VCAM-1、ICAM-1、p-ERK1/2和p-NF-κB蛋白表达水平降低（P<0.05）。房颤组大鼠心肌组织中MMP-2和MMP-9蛋白表达水平与VCAM-1（r=0.435，P<0.01；r=0.512，P<0.01）和ICAM-1（r=0.486，P<0.01；r=0.579，P<0.01）蛋白表达水平呈正相关关系。结论：丹参多酚酸可通过抑制房颤大鼠心房组织ERK1/2-NF-κB信号通路激活，降低VCAM-1和ICAM-1水平，抑制心肌纤维化，从而发挥对房颤的治疗作用。

关键词: 丹参多酚酸, 心房颤动, 血管细胞黏附分子1, 细胞间黏附分子1, 细胞外信号调节酶1/2-核转录因子-κB信号通路, 基质金属蛋白酶

Abstract: Objective: To study the effects of salvianolic acid on the levels of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) in myocardium tissue of atrial fibrillation rats and extracellular signal-regulating enzymes 1/2-nuclear transcription factor-κB (ERK1/2-NF-κB) signaling pathway, and to explore the possible mechanism of salvianolic acid in the preventing and treating atrial fibrillation. Methods: Forty-eight SD rats were randomly divided into control group, atrial fibrillation group and salvianolic acid group, with 16 rats in each group.The rat models of atrial fibrillation were established by intravenous injection of calcium chloride-acetylcholine mixture in the sublingual vein. The rats in salvianolic acid group were administrated with Salvianolic acid (4 mg·kg^-1·d^-1) by gavage and the rats in control group and atrial fibrillation group were intragastrically administered with an equal amount of normal saline, once a day, for 4 weeks. Hematoxylin-Eosin(HE) staining was used to observe the histopathological changes of myocardium tissue of the rats. Masson staining was used to observe the myocardial fibrosis. The expression levels of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9) proteins in myocardium tissue of the rats in various groups were measured by immunohistochemical staining. Western bloting method was used to determine the expression levels of VCAM-1, ICAM-1, ERK1/2, phosphorylated ERK1/2(p-ERK1/2), NF-κB and phosphorylated NF-κB (p-NF-κB) in myocardium tissue of the rats in various groups. Results: The HE staining results showed that there was no obvious abnormality in the myocardial cells of the rats in control group. In atrial fibrillation group, myocardial interstitial tissue was increased, and inflammatory cell infiltration was found. Compared with atrial fibrillation group,the myocardial interstitial tissue in salvianolic acid group was slightly more, and inflammatory cell infiltration was not obvious. The Masson staining results showed that the myocardial interstitial collagen fibers of the rats in control group were normal, and the myocardial interstitial collagen fibers of the rats in atrial fibrillation group were increased significantly; compared with atrial fibrillation group, the myocardial interstitial collagen fibers of the rats in salvianolic acid group were significantly reduced. Compared with control group, the expression levels of MMP-2, MMP-9,VCAM-1, ICAM-1, p-ERK1/2 and p-NF-κB proteins in myocardium tissue of the rats in atrial fibrillation group and salvianolic acid group were increased (P<0.05). Compared with atrial fibrillation group, the expression levels of MMP-2, MMP-9, VCAM-1, ICAM-1, p-ERK1/2 and p-NF-κB proteins in myocardium tissue of the rats in salvianolic acid group were reduced (P<0.05). The expression levels of MMP-2 and MMP-9 proteins in myocardium tissue of the rats in atrial fibrillation group were positively correlated with the expression levels of VCAM-1(r=0.435, P<0.01;r=0.512, P<0.05) and ICAM-1(r=0.486,P<0.01;r=0.579, P<0.01) proteins. Conclusion: Salvianolic acid can reduce the levels of VCAM-1 and ICAM-1, inhibit myocardial fibrosis, and exert the protective effect on atrial fibrillation by inhibiting the activation of ERK1/2-NF-κB signaling pathway in myocardium tissue of the atrial fibrillation rats.

Key words: salvianolic acid, atrial fibrillation, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, extracellular signal-regulating enzymes1/2-nuclear transcription factor-κB signaling pathway, matrix metalloproteinase

中图分类号:

R541.75

李小兵, 王旭, 吕瑛, 黄建成, 李红英, 王军, 张会军, 苏振宇. 丹参多酚酸对心房颤动大鼠心肌组织中VCAM-1和ICAM-1水平及ERK1/2-NF-κB信号通路的影响[J]. 吉林大学学报(医学版), 2020, 46(05): 1004-1010.

LI Xiaobing, WANG Xu, LYU Ying, HUANG Jiancheng, LI Hongying, WANG Jun, ZHANG Huijun, SU Zhenyu. Effects of salvianolic acid on levels of VCAM-1 and ICAM-1 in myocardium tissue and ERK1/2-NF-κB signaling pathway of atrial fibrillation rats[J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 1004-1010.

参考文献

[1] VAN WAGONER D R, CHUNG M K. Inflammation, inflammasome activation, and atrial fibrillation[J]. Circulation, 2018, 138(20):2243-2246.
[2] YAO C X, VELEVA T, SCOTT L, et al. Enhanced cardiomyocyte NLRP3 inflammasome signaling promotes atrial fibrillation[J]. Circulation, 2018, 138(20):2227-2242.
[3] KORANTZOPOULOS P, LETSAS K P, TSE G, et al. Inflammation and atrial fibrillation:A comprehensive review[J]. J Arrhythm, 2018, 34(4):394-401.
[4] WANG W, HU W. Salvianolic acid B recovers cognitive deficits and angiogenesis in a cerebral small vessel disease rat model via the STAT3/VEGF signaling pathway[J].Mol Med Rep, 2018,17(2):3146-3151.
[5] SONG Y, LIU W H, DING Y, et al. Salvianolic acid A ameliorates renal ischemia/reperfusion injury by activating Akt/mTOR/4EBP1 signaling pathway[J]. Am J Physiol Ren Physiol, 2018, 315(2):F254-F262.
[6] 刘维琴, 李卫松, 马清华. 丹酚酸对房颤大鼠TNF-α,MMP-2/TIMP-2表达影响随机平行对照研究[J]. 实用中医内科杂志, 2013,27(6):83-85.
[7] 何婉霞, 朱冠男. 高通量血液透析联合丹参多酚酸盐注射液对糖尿病肾病患者氧化应激及微炎症状态的影响[J]. 现代中西医结合杂志, 2019, 28(11):1207-1210.
[8] 李馨, 王文斌, 曹树军, 等. 丹参多酚酸盐辅助对不稳定型心绞痛患者TNF-α、hs-CRP、IL-1和心功能的影响[J]. 临床和实验医学杂志, 2018, 17(5):499-503.
[9] 陈春林, 巩甜甜, 汤依群, 等. SD大鼠房颤模型的建立[J]. 实验动物科学, 2009, 26(3):1-4.
[10] 刘维琴, 刘维蓉, 牟霞. 丹酚酸对房颤大鼠TNF-α,LTCCα1c蛋白表达影响随机平行对照研究[J]. 实用中医内科杂志, 2013, 27(2):65-68.
[11] SONG F C, JI B, CHEN T. Cilostazol on the expression of ICAM-1, VCAM-1 and inflammatory factors in plasma in patients with thromboangiitis obliterans[J]. Exp Ther Med, 2018, 16(3):2349-2354.
[12] HSU S F, LEE Y B, LEE Y C, et al. Dual specificity phosphatase DUSP6 promotes endothelial inflammation through inducible expression of ICAM-1[J]. FEBS J, 2018, 285(9):1593-1610.
[13] RUEDIGER C D, JOHN B, KUMAR S, et al. Influence of ethnic background on left atrial markers of inflammation, endothelial function and tissue remodelling[J]. Indian Pacing Electrophysiol J, 2018, 18(1):1-5.
[14] WILLEIT K, PECHLANER R, WILLEIT P, et al. Association between vascular cell adhesion molecule 1 and atrial fibrillation[J]. JAMA Cardiol, 2017, 2(5):516-523.
[15] CHIN J P, CHEN C M, LEE T H, et al. Angiostrongylus cantonensis-conditioned culture medium induces myelin basic protein alterations via Erk1/2 and NF-κB activation in rat RSC96 schwann cells[J]. Chin J Physiol, 2018, 61(3):137-143.
[16] CHENG L J, CHEN Z H, WANG L H, et al. Propofol partially attenuates complete freund's adjuvant-induced neuroinflammation through inhibition of the ERK1/2/NF-κB pathway[J]. J Cell Biochem, 2019, 120(6):9400-9408.
[17] ZHONG L, SIMARD M J, HUOT J. Endothelial microRNAs regulating the NF-κB pathway and cell adhesion molecules during inflammation[J]. FASEB J, 2018, 32(8):4070-4084.
[18] OLIVARES S F, LANDAET R, ARANGUIZ P, et al. Heparan sulfate potentiates leukocyte adhesion on cardiac fibroblast by enhancing VCAM-1 and ICAM-1 expression[J]. Biochim Biophys Acta Mol Basis Dis, 2018, 1864(3):831-842.
[19] SUN Y, ZHAO D L, LIU Z X, et al. Inhibitory effect of salvianolic acid on inflammatory mediators of rats with collagen-induced rheumatoid arthritis[J]. Exp Ther Med, 2018, 16(5):4037-4041.
[20] ZHANG H F, WANG Y L, GAO C, et al. Salvianolic acid A attenuates kidney injury and inflammation by inhibiting NF-κB and p38 MAPK signaling pathways in 5/6 nephrectomized rats[J]. Acta Pharmacol Sin, 2018, 39(12):1855-1864.
[21] ABE I, TESHIMA Y, KONDO H, et al. Association of fibrotic remodeling and cytokines/chemokines content in epicardial adipose tissue with atrial myocardial fibrosis in patients with atrial fibrillation[J]. Heart Rhythm,2018, 15(11):1717-1727.
[22] XUE Y M, DENG C Y, WEI W, et al. Macrophage migration inhibitory factor promotes cardiac fibroblast proliferation through the Src kinase signaling pathway[J]. Mol Med Rep, 2018, 17(2):3425-3431.
[23] 王栋, 白雪蕾, 吴环立. 急性期脑出血患者血清MMP-2 ICAM-1与IL-6 TNF-α水平和脑水肿的关系[J]. 中国实用神经疾病杂志, 2019, 22(12):1336-1341.
[24] AHMED S, RIEGSECKER S, BEAMER M, et al. Largazole, a class I histone deacetylase inhibitor, enhances TNF-α-induced ICAM-1 and VCAM-1 expression in rheumatoid arthritis synovial fibroblasts[J]. Toxicol Appl Pharmacol, 2013, 270(2):87-96.