Zhang Meiqi, Cheng Kang, Yu Limei, Wu Weihua, Wang Yakun, Chen Yun
Hangzhou Traditional Chinese Medicine Hospital Affiliated to Zhejiang Chinese Medical University, Hangzhou 310021, Zhejiang, China. Corresponding author: Chen Yun, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2022 Jun;34(6):640-645. doi: 10.3760/cma.j.cn121430-20220309-00232.
To explore the effect of tanshinone II A on myocardial remodeling in ischemia/reperfusion (I/R)-induced heart failure of rodent model.
(1) In vivo, 30 SD rats were randomly divided into sham operation, heart failure and tanshinone II A treatment group, with 10 rats in each group. The I/R model was established by ligating the left coronary artery until ST segment elevation for 30 minutes, then the ligation was removed for 2 hours as reperfusion. In the sham operation group, the rat chest was opened without artery ligation. Three days after model establishment, tanshinone II A (10 mg/kg) were given intraperitoneal injected in tanshinone II A group for 9 weeks. In the other two groups, normal saline was administrated in the same way. The behavioral manifestations of the rats in each group were observed; hemodynamic indexes were evaluated; Masson staining was performed to observe the degree of myocardial fibrosis; enzyme linked immunosorbent assay (ELISA) was used to detect the content of Galectin-3 in myocardial tissue; quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expressions of collagen III, collagen I, matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase (TIMP-1). (2) In vitro, rats primary cardiac fibroblasts were extracted and isolated, and divided into blank control group, angiotensin II group (7-10 mmol/L angiotensin II) and angiotensin II + tanshinone II A group (7-10 mmol/L angiotensin II + 5-10 mmol/L tanshinone II A). At 24 hours and 48 hours of culture, the cell proliferation in each group was detected by methyl thiazolyl tetrazolium (MTT); the expressions of collagen III, collagen I, MMP-2 and TIMP-1 were detected by qRT-PCR; the content of Galectin-3 in cardiac fibroblasts was detected by ELSIA.
(1) In vivo, the rats' activity status, hair conformity and food intake were ranked from good to bad in order of sham operation group, tanshinone II A group and heart failure model group. Compared with the sham-operated group, the heart rate (HR) of the rats in the heart failure model group was significantly decreased and the heart function was significantly impaired. The mRNA and protein expression of collagen I, collagen III, TIMP-1 and Galectin-3 content were significantly increased, while the mRNA and protein expression of MMP-2 were significantly decreased. Compared with the heart failure model group, rats in the tanshinone II A group showed significantly higher HR and improved cardiac function, significantly lower mRNA expression of collagen I and collagen III, significantly lower mRNA and protein expression of TIMP-1 and Galectin-3, and significantly higher mRNA and protein expression of MMP-2, and the most obvious changes were in the 9th weeks of modeling [collagen I mRNA (2): 4.70±1.19 vs. 10.21±1.62, collagen III mRNA (2): 3.03±0.46 vs. 13.84±1.93, TIMP-1 mRNA (2): 1.90±0.19 vs. 4.55±0.43, TIMP-1/GAPDH: 0.33±0.04 vs. 0.67±0.05, Galectin-3 (ng/L): 489.93±79.30 vs. 821.72±94.09, MMP-2 mRNA (2): 0.37±0.07 vs. 0.03±0.01, MMP-2/GAPDH: 0.69±0.09 vs. 0.21±0.04, all P < 0.05]. Masson staining showed that myocardial tissue fibrosis was obvious in the heart failure group, and the degree of fibrosis in the tanshinone II A group was reduced. (2) In vitro, compared with the blank control group, the proliferation rate, collagen I, collagen III and TIMP-1 expression and Galectin-3 content of myocardial fibroblasts were significantly increased, and MMP-2 expression was significantly decreased in the angiotensin group at 24 h and 48 h of culture. Compared with the angiotensin group, the proliferation rate of cardiac fibroblasts and the expression of collagen I, collagen III and TIMP-1 and the content of Galectin-3 were significantly decreased, and the expression of MMP-2 mRNA was significantly increased in the angiotensin + tanshinone II A group, and the most significant changes were at 48 hours of culture [proliferation rate: (57.0±3.7)% vs. (67.0±2.4)%, collagen I mRNA (2): 551.43±67.10 vs. 871.48±12.25, collagen III mRNA (2): 233.76±18.73 vs. 385.51±31.35, TIMP-1 mRNA (2): 238.69±17.37 vs. 351.84±26.17, Galectin-3 (ng/L): 283.76±28.73 vs. 415.51±31.35, MMP-2 mRNA (2): 108.54±12.10 vs. 51.47±6.25, all P < 0.05].
Tanshinone II A can improve cardiac function, inhibit myocardial fibrosis and improve myocardial remodeling in rats with I/R-induced heart failure.
探讨丹参酮II A对缺血/再灌注（I/R）诱导的啮齿动物模型心力衰竭心肌重构的影响。
（1）体内实验，将30只SD大鼠随机分为假手术组、心力衰竭组和丹参酮II A治疗组，每组10只。通过结扎左冠状动脉直至ST段抬高30分钟建立I/R模型，然后松开结扎进行2小时再灌注。假手术组大鼠打开胸腔但不结扎动脉。造模3天后，丹参酮II A组腹腔注射丹参酮II A（10 mg/kg），持续9周。其他两组以同样方式给予生理盐水。观察每组大鼠的行为表现；评估血流动力学指标；进行Masson染色观察心肌纤维化程度；采用酶联免疫吸附测定（ELISA）检测心肌组织中半乳糖凝集素-3的含量；采用定量逆转录-聚合酶链反应（qRT-PCR）检测III型胶原、I型胶原、基质金属蛋白酶2（MMP-2）和金属蛋白酶组织抑制剂（TIMP-1）的表达。（2）体外实验，提取并分离大鼠原代心肌成纤维细胞，分为空白对照组、血管紧张素II组（7 - 10 mmol/L血管紧张素II）和血管紧张素II + 丹参酮II A组（7 - 10 mmol/L血管紧张素II + 5 - 10 mmol/L丹参酮II A）。培养24小时和48小时时，采用噻唑蓝（MTT）法检测每组细胞增殖情况；采用qRT-PCR检测III型胶原、I型胶原、MMP-2和TIMP-1的表达；采用ELISA检测心肌成纤维细胞中半乳糖凝集素-3的含量。
（1）体内实验，大鼠的活动状态、毛发状态和进食量按假手术组、丹参酮II A组和心力衰竭模型组顺序由好到差。与假手术组相比，心力衰竭模型组大鼠心率（HR）显著降低，心功能显著受损。I型胶原、III型胶原、TIMP-1的mRNA和蛋白表达及半乳糖凝集素-3含量显著增加，而MMP-2的mRNA和蛋白表达显著降低。与心力衰竭模型组相比，丹参酮II A组大鼠HR显著升高，心功能改善，I型胶原和III型胶原的mRNA表达显著降低，TIMP-1和半乳糖凝集素-3的mRNA和蛋白表达显著降低，MMP-2的mRNA和蛋白表达显著升高，建模第9周变化最明显[I型胶原mRNA（2）：4.70±1.19 vs. 10.21±1.62，III型胶原mRNA（2）：3.03±0.46 vs. 13.84±1.93，TIMP-1 mRNA（2）：1.90±0.19 vs. 4.55±0.43，TIMP-1/GAPDH：0.33±0.04 vs. 0.67±0.05，半乳糖凝集素-3（ng/L）：489.93±79.30 vs. 821.72±94.09，MMP-2 mRNA（2）：0.37±0.07 vs. 0.03±0.01，MMP-2/GAPDH：0.69±0.09 vs. 0.21±0.04，均P < 0.05]。Masson染色显示心力衰竭组心肌组织纤维化明显，丹参酮II A组纤维化程度减轻。（2）体外实验，与空白对照组相比，血管紧张素组培养24小时和48小时时心肌成纤维细胞的增殖率、I型胶原、III型胶原和TIMP-1表达及半乳糖凝集素-3含量显著增加，MMP-2表达显著降低。与血管紧张素组相比，血管紧张素 + 丹参酮II A组心肌成纤维细胞的增殖率及I型胶原、III型胶原和TIMP-1表达和半乳糖凝集素-3含量显著降低，MMP-2 mRNA表达显著增加，培养48小时时变化最显著[增殖率：（57.0±3.7）% vs. （67.0±2.4）%，I型胶原mRNA（2）：551.43±67.10 vs. 871.48±12.25，III型胶原mRNA（2）：233.76±18.73 vs. 385.51±31.35，TIMP-1 mRNA（2）：238.69±17.37 vs. 351.84±26.17，半乳糖凝集素-3（ng/L）：283.76±28.73 vs. 415.51±31.35，MMP-2 mRNA（2）：108.54±12.10 vs. 51.47±6.25，均P < 0.05]。
丹参酮II A可改善I/R诱导的心力衰竭大鼠的心功能，抑制心肌纤维化，改善心肌重构。
