丹参酮ⅡA通过抑制NADPH氧化酶保护心衰大鼠心肌纤维化的机制
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摘要
目的探讨丹参酮ⅡA抑制还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶改善心肌纤维化的作用机制。方法利用Anversea方法制备心衰大鼠模型,将28只雌性大鼠随机分成4组,对照组、丹参酮ⅡA组、纤维化模型组、纤维化+丹参酮ⅡA组。对照组、丹参酮ⅡA组只分离腹主动脉,不造成动脉狭窄;丹参酮ⅡA组、纤维化+丹参酮ⅡA组每日上午经腹腔注射给予15 mg/kg的丹参酮ⅡA。检测各组大鼠心肌NADPH氧化酶活性、心肌超氧化物歧化酶(SOD)含量、心肌丙二醛(MDA)含量、Nox2及Nox4的mRNA水平表达、心肌胶原含量。结果心衰时大鼠心肌中NADPH氧化酶活性增强,达到(14.07±0.23) nmol/mg;丹参酮ⅡA注射后,NADPH氧化酶活性降到(8.72±0.28) nmol/mg;大鼠心衰时,心肌组织中SOD活力下降,MDA含量增高;纤维化+丹参酮ⅡA组的SOD活力优于纤维化模型组,纤维化模型组MDA含量高于纤维化+丹参酮ⅡA组;丹参酮ⅡA注射后,可提高SOD的活性;纤维化模型组和纤维化+丹参酮ⅡA组大鼠的Nox2 mRNA水平无明显变化,纤维化模型组和纤维化+丹参酮ⅡA组大鼠的Nox4 mRNA出现显著增加;纤维化模型组大鼠的心肌胶原容积百分比显著提高,心肌纤维化程度提高到32.44%,远高于对照组的5.13%,纤维化+丹参酮ⅡA组的胶原容积百分比为18.37%。结论丹参酮ⅡA注射抑制心衰大鼠心肌NADPH氧化酶中Nox4的表达,抑制了心肌氧化应激水平,同时丹参酮ⅡA升高SOD活力,从而抑制了心衰大鼠心肌纤维化水平。
Objective To investigate the mechanism of tanshinone ⅡA inhibiting NADPH oxidase in improving myocardial fibrosis. Methods The rat model of heart failure was prepared by Anversea, and 28 female rats were randomly divided into 4 groups, the control group, the tanshinone ⅡA group, the fibrosis model group, and the fibrosis + tanshinone ⅡA group. The control group and tanshinone ⅡA group only separated the abdominal aorta and did not cause arterial stenosis, the tanshinone ⅡA group, and the fibrosis + tanshinone ⅡA group were given 15 mg/kg tanshinone ⅡA every morning by intraperitoneal injection. The mechanism of tanshinone ⅡA in improving myocardial fibrosis was comprehensively assessed by macroscopic characterization combined with physical and chemical indicators. The myocardial NADPH oxidase activity, myocardial SOD content, myocardial malondialdehyde(MDA) content, Nox2, Nox4 mRNA level and myocardial collagen content were detected. Results The activity of NADPH oxidase in myocardium of heart failure rats increased to(14.07±0.23) nmol/mg. After injection of tanshinone ⅡA, the activity of NADPH oxidase decreased to(8.72±0.28) nmol/mg. In heart failure(HF) rats, SOD activity in myocardium decreased and MDA content increased, SOD activity in fibrosis + tanshinone ⅡA group was better than that in fibrosis model group, and MDA content in fibrosis model group was higher than that in fibrosis + tanshinone ⅡA group. Tanshinone ⅡA injection can increase the activity of SOD. There was no significant change in the level of Nox2 mRNA in the fibrosis model group and the fibrosis + tanshinone ⅡA group, but the Nox4 mRNA in the fibrosis model group and the fibrosis + tanshinone ⅡA group increased significantly. The percentage of myocardial collagen volume in fibrosis model group was significantly increased to 32.44%, which was much higher than that in control group(5.13%). The percentage of collagen volume in fibrosis + tanshinone ⅡA group was 18.37%. Conclusion Tanshinone ⅡA injection inhibits the expression of Nox4 in myocardial NADPH oxidase in heart failure rats and inhibits the level of myocardial oxidative stress, while tanshinone ⅡA increases SOD activity, thus inhibiting the level of myocardial fibrosis in rats with heart failure.
Objective To investigate the mechanism of tanshinone ⅡA inhibiting NADPH oxidase in improving myocardial fibrosis. Methods The rat model of heart failure was prepared by Anversea, and 28 female rats were randomly divided into 4 groups, the control group, the tanshinone ⅡA group, the fibrosis model group, and the fibrosis + tanshinone ⅡA group. The control group and tanshinone ⅡA group only separated the abdominal aorta and did not cause arterial stenosis, the tanshinone ⅡA group, and the fibrosis + tanshinone ⅡA group were given 15 mg/kg tanshinone ⅡA every morning by intraperitoneal injection. The mechanism of tanshinone ⅡA in improving myocardial fibrosis was comprehensively assessed by macroscopic characterization combined with physical and chemical indicators. The myocardial NADPH oxidase activity, myocardial SOD content, myocardial malondialdehyde(MDA) content, Nox2, Nox4 mRNA level and myocardial collagen content were detected. Results The activity of NADPH oxidase in myocardium of heart failure rats increased to(14.07±0.23) nmol/mg. After injection of tanshinone ⅡA, the activity of NADPH oxidase decreased to(8.72±0.28) nmol/mg. In heart failure(HF) rats, SOD activity in myocardium decreased and MDA content increased, SOD activity in fibrosis + tanshinone ⅡA group was better than that in fibrosis model group, and MDA content in fibrosis model group was higher than that in fibrosis + tanshinone ⅡA group. Tanshinone ⅡA injection can increase the activity of SOD. There was no significant change in the level of Nox2 mRNA in the fibrosis model group and the fibrosis + tanshinone ⅡA group, but the Nox4 mRNA in the fibrosis model group and the fibrosis + tanshinone ⅡA group increased significantly. The percentage of myocardial collagen volume in fibrosis model group was significantly increased to 32.44%, which was much higher than that in control group(5.13%). The percentage of collagen volume in fibrosis + tanshinone ⅡA group was 18.37%. Conclusion Tanshinone ⅡA injection inhibits the expression of Nox4 in myocardial NADPH oxidase in heart failure rats and inhibits the level of myocardial oxidative stress, while tanshinone ⅡA increases SOD activity, thus inhibiting the level of myocardial fibrosis in rats with heart failure.
引文
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[8] 戴新新, 宿树兰, 郭盛,等. 丹参酮类成分的生物活性与应用开发研究进展[J]. 中草药, 2017, 48(7):1442-8.
[9] Maione F, Piccolo M, De Vita S, et al. Down regulation of pro-inflammatory pathways by tanshinone IIA and cryptotanshinone in a non-genetic mouse model of Alzheimer?s disease[J]. Pharmacol Res, 2012, 1299(3):482-90.
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[11] Wang S, Jing H, Yang H, et al. Tanshinone I selectively suppresses pro-inflammatory genes expression in activated microglia and prevents nigrostriatal dopaminergic neurodegeneration in a mouse model of Parkinson?s disease[J]. J Ethnopharmacol, 2015, 164(8):247-55.
[12] 王汉章. ERK1/2信号通路介导PDGF-CC诱导的鼠心肌成纤维细胞增殖和胶原合成的分子机制[D].合肥:安徽医科大学, 2017.
[13] 于婷, 王小梅. 通心络对高糖培养下心肌成纤维细胞增殖和凋亡的影响[J]. 中国医科大学学报, 2018, 47(2):132-6.
[14] 程琪,李勇文,李丽, 等.甲基阿魏酸对乙醇诱导HSC-LX2 细胞NOX4 和α-SMA 表达的影响[J].安徽医科大学学报, 2018, 53(4):509-15.
[2] Porwol T, Ehleben W, Brand V, et al. Tissue oxygen sensor function of NADPH oxidase isoforms, an unusual cytochrome aa3 and reactive oxygen species[J].Respir Physiol, 2001, 128(3):331-48.
[3] Huo Y, Rangarajan P, Ling E A, et al. Dexamethasone inhibits the Nox-dependent ROS production via suppression of MKP-1-dependent MAPK pathways in activated microglia[J]. Bmc Neuroscience, 2011, 12(1):49.
[4] 贺环宇. 丹参酮ⅡA对大鼠肺间质纤维化的保护作用及机制研究[D].大连:大连医科大学, 2014.
[5] Burgoyne J R, Monguedin H, Eaton P, et al. Redox signaling in cardiac physiology and pathology[J]. Circ Res, 2012, 111(8):1091-106.
[6] 解华. 基于NADPH氧化酶活性调控的益心解毒方治疗心力衰竭的分子机制研究[D]. 北京:北京中医药大学, 2015.
[7] 房晓楠, 张荣成, 张健. T细胞免疫调节机制在心力衰竭心肌纤维化中的作用[J]. 中国循环杂志, 2015, 30(5):508-10.
[8] 戴新新, 宿树兰, 郭盛,等. 丹参酮类成分的生物活性与应用开发研究进展[J]. 中草药, 2017, 48(7):1442-8.
[9] Maione F, Piccolo M, De Vita S, et al. Down regulation of pro-inflammatory pathways by tanshinone IIA and cryptotanshinone in a non-genetic mouse model of Alzheimer?s disease[J]. Pharmacol Res, 2012, 1299(3):482-90.
[10] Yu Z L, Wang J N, Wu X H, et al. Tanshinone IIA prevents rat basilar artery smooth muscle cells proliferation by inactivation of PDK1 during the development of hypertension[J]. J Cardiovasc Pharmacol Ther, 2018, 20(6):563-71.
[11] Wang S, Jing H, Yang H, et al. Tanshinone I selectively suppresses pro-inflammatory genes expression in activated microglia and prevents nigrostriatal dopaminergic neurodegeneration in a mouse model of Parkinson?s disease[J]. J Ethnopharmacol, 2015, 164(8):247-55.
[12] 王汉章. ERK1/2信号通路介导PDGF-CC诱导的鼠心肌成纤维细胞增殖和胶原合成的分子机制[D].合肥:安徽医科大学, 2017.
[13] 于婷, 王小梅. 通心络对高糖培养下心肌成纤维细胞增殖和凋亡的影响[J]. 中国医科大学学报, 2018, 47(2):132-6.
[14] 程琪,李勇文,李丽, 等.甲基阿魏酸对乙醇诱导HSC-LX2 细胞NOX4 和α-SMA 表达的影响[J].安徽医科大学学报, 2018, 53(4):509-15.