玉郎伞MHBFC调控eNOS-NO信号通路逆转大鼠心室重构的机制研究
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摘要
目的观察玉郎伞单体17-甲氧基-7-羟基-苯并呋喃查尔酮(MHBFC)调控心肌微血管内皮细胞(MMVEC)eNOS-NO信号通路,逆转大鼠压力超负荷心室重构的机制。方法将缩窄腹主动脉的成活大鼠随机分为假手术组、模型组、MHBFC 6 mg·kg~(-1)组、MHBFC 12 mg·kg~(-1)组、MHBFC 12 mg·kg~(-1)+亚硝基左旋精氨酸甲酯(L-NAME)50 mg·kg~(-1)组、L-NAME 50 mg·kg~(-1)组。假手术组只游离腹主动脉,不缩窄。分组后,各组给予相应药物6周。化学消化法分离左心室组织获得MMVEC;Dil-Ac-LDL吞噬实验鉴定MMVEC;qPCR检测MMVEC eNOS、PI3K和Akt基因表达;Western blot检测MMVEC p-eNOS、p-PI3K和p-Akt蛋白表达。结果 MHBFC预处理组明显上调MMVEC p-eNOS、p-PI3K、p-Akt蛋白表达,以及MMVEC eNOS、PI3K、Akt基因表达;L-NAME组MMVEC p-eNOS蛋白及eNOS基因表达明显下调;合用MHBFC后明显逆转上述指标的下调。结论 MHBFC可能通过增加MMVEC PI3K、Akt磷酸化及PI3K、Akt基因表达,增加MMVEC eNOS蛋白磷酸化及eNOS基因表达,激活MMVEC eNOS-NO信号通路,从而逆转压力超负荷心室重构。
Aim To observe the mechanism of Yulangsan monomers 17-methoxyl-7-hydroxy-benzene-furanchalcone(MHBFC) regulating myocardial microvascular endothelial cell(MMVEC) eNOS-NO signaling pathway in reversing pressure overload-induced ventricular remodeling rats. Methods The survival rats with narrowed abdominal aorta were randomly divided into sham operated group, model group, MHBFC 6 mg·kg~(-1) group, MHBFC 12 mg·kg~(-1) group, MHBFC 12 mg·kg~(-1) + Nω-nitro-L-arginine methyl ester hydrochloride(L-NAME) 50 mg·kg~(-1) group and L-NAME 50 mg·kg~(-1) group. In the sham operated group, only the abdominal aorta was free and not narrowed. After grouping, each group was given the corresponding drug for six weeks. MMVEC was obtained by chemical digestion in left ventricular tissue, and MMVEC was identified by Dil-Ac-LDL phagocytosis. The gene expression of MMVEC eNOS, PI3 K and Akt was detected by qPCR. The protein expression of MMVEC p-eNOS, p-PI3 K and p-Akt was detected using Western blot. Results The protein expression of MMVEC p-eNOS, p-PI3 K and p-Akt and the gene expression of MMVEC eNOS, PI3 K and Akt significantly increased in MHBFC pretreatment group. The expression of MMVEC p-eNOS protein and the expression of eNOS gene were obviously down-regulated in L-NAME group, and the down-regulation of the above index was obviously reversed after the combination of MHBFC. Conclusions MHBFC may increase the phosphorylation of MMVEC eNOS protein and the expression of MMVEC eNOS gene by increasing the phosphorylation of PI3 K and Akt proteins and the expression of PI3 K and Akt genes in MMVEC, and activate the eNOS-NO signaling pathway in MMVEC, thereby reversing pressure overload-induced ventricular remodeling.
Aim To observe the mechanism of Yulangsan monomers 17-methoxyl-7-hydroxy-benzene-furanchalcone(MHBFC) regulating myocardial microvascular endothelial cell(MMVEC) eNOS-NO signaling pathway in reversing pressure overload-induced ventricular remodeling rats. Methods The survival rats with narrowed abdominal aorta were randomly divided into sham operated group, model group, MHBFC 6 mg·kg~(-1) group, MHBFC 12 mg·kg~(-1) group, MHBFC 12 mg·kg~(-1) + Nω-nitro-L-arginine methyl ester hydrochloride(L-NAME) 50 mg·kg~(-1) group and L-NAME 50 mg·kg~(-1) group. In the sham operated group, only the abdominal aorta was free and not narrowed. After grouping, each group was given the corresponding drug for six weeks. MMVEC was obtained by chemical digestion in left ventricular tissue, and MMVEC was identified by Dil-Ac-LDL phagocytosis. The gene expression of MMVEC eNOS, PI3 K and Akt was detected by qPCR. The protein expression of MMVEC p-eNOS, p-PI3 K and p-Akt was detected using Western blot. Results The protein expression of MMVEC p-eNOS, p-PI3 K and p-Akt and the gene expression of MMVEC eNOS, PI3 K and Akt significantly increased in MHBFC pretreatment group. The expression of MMVEC p-eNOS protein and the expression of eNOS gene were obviously down-regulated in L-NAME group, and the down-regulation of the above index was obviously reversed after the combination of MHBFC. Conclusions MHBFC may increase the phosphorylation of MMVEC eNOS protein and the expression of MMVEC eNOS gene by increasing the phosphorylation of PI3 K and Akt proteins and the expression of PI3 K and Akt genes in MMVEC, and activate the eNOS-NO signaling pathway in MMVEC, thereby reversing pressure overload-induced ventricular remodeling.
引文
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[13] Yu Y, Wang H Y, Yi Z P, et al. Improvement effects of parecoxib on ventricular remodeling after acute myocardial infarction in rats and its influence in PI3K/Akt signaling pathway [J]. Chin Pharmacol Bull, 2017, 33(6): 863-8.
[14] Chang W, Wu Q Q, Xiao Y, et al. Acacetin protects against cardiac remodeling after myocardial infarction by mediating MAPK and PI3K/Akt signal pathway [J]. J Pharmacol Sci, 2017, 135(4): 156-63.
[2] Byrne N J, Levasseur J, Sung M M, et al. Normalization of cardiac substrate utilization and left ventricular hypertrophy precede functional recovery in heart failure regression[J]. Cardiovasc Res, 2016, 110(2): 249-57.
[3] 简 洁, 黄建春, 焦 杨, 等. 制备液相色谱分离制备玉郎伞查尔酮类化合物[J]. 中草药, 2011, 42(7): 1313-6.
[3] Jian J, Huang J C, Jiao Y, et al. Isolation and preparation of chalcone compounds from tuber of Millettia pulchra var. laxior by pre-HPLC [J]. Chin Tradit Herb Drugs, 2011, 42(7): 1313-6.
[4] Huang J, Tang X, Liang X, et al. The effects of 17-methoxyl-7-hydroxy-benzene-furanchalcone on the pressure overload-induced progression of cardiac hypertrophy to cardiac failure[J]. PLoS One, 2014, 9(3): e91834.
[5] Feng B, Zhang Q, Wang X, et al. Effect of andrographolide on gene expression profile and intracellular calcium in primary rat myocardium microvascular endothelial cells [J]. J Cardiovasc Pharmacol, 2017, 70(6): 369-81.
[6] 辛 毅, 许秀芳, 黄益民, 等. 小鼠心肌微血管内皮细胞的原代培养及生物学特性[J]. 中国病理生理杂志, 2013, 29(3): 565-70.
[6] Xin Y, Xu X F, Huang Y M, et al. Biological characteristics of primarily cultured microvascular endothelial cells in mouse myocardium [J]. Chin J Pathophysiol, 2013, 29(3): 565-70.
[7] Lewalle A, Land S, Carruth E, et al. Decreasing compensatory ability of concentric ventricular hypertrophy in aortic-banded rat hearts [J]. Front Physiol, 2018, 9: 37.
[8] 张 伟, 刘晓丹, 李 菲, 等. 血府逐瘀汤调控内皮祖细胞修复损伤血管内皮的研究[J]. 中国药理学通报, 2016, 32(3): 427-33.
[8] Zhang W, Liu X D, Li F, et al. Influence of Xuefuzhuyu decoction on EPC repairing injured vascular endothelium [J]. Chin Pharmacol Bull, 2016, 32(3): 427-33.
[9] Zhou Q, Meng G, Teng F, et al. Effects of astragalus polysaccharide on apoptosis of myocardial microvascular endothelial cells in rats undergoing hypoxia/reoxygenation by mediation of the PI3K/Akt/eNOS signaling pathway [J]. J Cell Biochem, 2018, 119(1): 806-16.
[10] Bunbupha S, Prachaney P, Kukongviriyapan U, et al. Asiatic acid alleviates cardiovascular remodelling in rats with L-NAME-induced hypertension [J]. Clin Exp Pharmacol Physiol, 2015, 42(11): 1189-97.
[11] 付剑江, 王曦聆, 吕 红, 等. 罗布麻叶提取物的抗高血压作用及其机制研究[J]. 中国实验方剂学杂志, 2013, 19(7): 159-64.
[11] Fu J J, Wang X L, Lyu H, et al. Anti-hypertensive effects of extracts from leaves of Apocynum venetum through activating PI3K/Akt pathway on human endothelial cells [J]. Chin J Exp Tradit Med Form, 2013, 19(7): 159-64.
[12] Ou H C, Lee W J, Lee S D, et al. Ellagic acid protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/eNOS pathway[J]. Toxicol Appl Pharmacol, 2010, 248(2): 134-43.
[13] 余 娅, 王虹又, 易宗平, 等. 帕瑞昔布对大鼠心梗后心室重构的改善作用及对PI3K/Akt信号通路的影响 [J]. 中国药理学通报, 2017, 33(6): 863-8.
[13] Yu Y, Wang H Y, Yi Z P, et al. Improvement effects of parecoxib on ventricular remodeling after acute myocardial infarction in rats and its influence in PI3K/Akt signaling pathway [J]. Chin Pharmacol Bull, 2017, 33(6): 863-8.
[14] Chang W, Wu Q Q, Xiao Y, et al. Acacetin protects against cardiac remodeling after myocardial infarction by mediating MAPK and PI3K/Akt signal pathway [J]. J Pharmacol Sci, 2017, 135(4): 156-63.