摘要
目的探讨心舒宝对心肌梗死大鼠的保护作用及相关机制。方法 Wistar大鼠(250~300 g)随机分为6组:假手术组,模型组,心舒宝低、中、高剂量组和地奥心血康组。模型组结扎左冠状动脉前降支造成心肌梗死模型,假手术组除不结扎冠状动脉外,其余操作与模型组相同。术后1 d,心舒宝组分别给予37.5、75、150 mg/(kg·d)灌胃给药,地奥心血康组按照150 mg/(kg·d)灌胃给药,模型组和假手术组灌胃相同体积生理盐水,1次/d。连续给药4周后采用超声心动图系统和BL-420F生物机能实验系统测定心功能及血流动力学指标。然后取心脏切片,采用TTC染色观察心肌梗死面积以及HE染色法观察心肌细胞形态学变化。此外,利用试剂盒检测心脏组织中超氧化物歧化酶(Superoxide dismutase, SOD)、乳酸脱氢酶(lactate dehydrogenase, LDH)活性以及丙二醛(Malondialdehyde, MDA)、还原型谷胱甘肽(Glutathione, GSH)、白介素-18(interleukin-18,IL-18)、白介素1β(interleukin 1β,IL-1β)含量;采用Western Blot检测B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、Bax、Caspase 3的蛋白表达情况。结果与假手术组相比,模型组大鼠的心脏功能指标显著下降(P<0.01),伴随着梗死面积显著增大(P<0.01)。同时出现了大量的梗死灶,核消失,横纹消失,肌浆变成不规则颗粒状,肌纤维成条索状,梗死区炎症反应明显,大量炎症细胞浸润等症状。此外,SOD、LDH活性及还原型GSH含量显著降低(P<0.01),而MDA含量则显著升高(P<0.01),IL-18、IL-1β含量也显著升高(P<0.01);且Bcl-2表达显著下调,Bax、Caspase 3表达显著上调(P<0.01),表明大鼠心肌梗死模型制备成功。与模型组相比,心舒宝中、高剂量组及地奥心血康组大鼠的心脏功能指标显著上升(P<0.05,P<0.01),出现梗死面积显著减小(P<0.05,P<0.01),梗死灶明显变小,心肌坏死程度显著减轻,胞质呈现淡染,大部分心肌可见收缩带,炎症细胞浸润减小等恢复性症状。心舒宝中、高剂量组和地奥心血康组大鼠的SOD、LDH活性及还原型GSH含量显著升高,MDA含量显著降低(P<0.05,P<0.01)。心脏组织中IL-18、IL-1β含量显著降低(P<0.05,P<0.01),Bcl-2表达显著上调,Bax、Caspase 3的表达显著下调(P<0.05,P<0.01)。结论心舒宝对心肌梗死前降支结扎造成的心脏损伤具有保护作用,该保护作用与其抗炎和抗凋亡作用密切相关。
Objective To study the protective effect and mechanism of Xinshubao(心舒宝) on the heart of rats with myocardial infarction. Method Firstly, SD rats(250~300 g) were randomly divided into 6 groups: sham operation group, model group, Xinshubao low dose group, Xinshubao medium dose group, Xinshubao high dose group and Di’ao Xinxuekang group. In the model group, ligation of left anterior descending coronary artery resulted in the myocardial infarction model. In the sham operation group, the operation was identical with that of the model group except that no ligation of the coronary artery was performed. Three days after the operation, Xinshubao group was given 37.5, 75, 150 mg/(kg·d) respectively by intragastric administration and Di’ao Xinxuekang group was given at 150 mg/(kg·d) by intragastric administration for 4 weeks, once a day, while the model group and the sham operation group were fed normally. Next, echocardiography system and BL-420 F biological function experiment system were used to measure cardiac function and hemodynamics. Then, the heart was extracted, and the myocardial infarction area was observed with TTC. The morphological changes of myocardial cells were observed by HE staining. In addition, the activity of SOD and LDH as well as the content of MDA was reduced. GSH, IL-18 and IL-1β in the cardiac tissue were detected by the kit. Last, the expressions of Bcl-2, Bax and Caspase 3 were detected by Western blot. Result Compared with that of the sham operation group, the cardiac function index of the model group was significantly decreased(P<0.01), accompanied with a significant increase in infarct size(P<0.01). At the same time, a large number of infarcts appeared, while the nucleus and the horizontal stripes disappeared. The sarcoplasm became irregular granules, and the muscle fibers became cord-like. The inflammatory reaction in the infarct area was obvious, with a large number of neutrophil. In addition, the activities of SOD and LDH and the content of reduced GSH were significantly decreased(P<0.01), while the content of MDA and the levels of IL-18 and IL-1β were significantly increased(P<0.01). The expression of Bcl-2 was significantly down-regulated. Bax and Caspase 3 were significantly up-regulated(P<0.01). This indicated that the rat myocardial infarction model was successfully performed. On the other hand, compared with those of the model group, the cardiac function indexes of the Xinshubao medium and high dose groups and the Di’ao Xinxuekang group were significantly increased(P<0.05,P<0.01), while the infarct size was significantly decreased(P<0.05,P<0.01), and the infarct was significantly smaller. The degree of myocardial necrosis was significantly reduced, and the cytoplasm was lightly stained. At the same time, most of the myocardium showed signs of contraction with reduced neutrophil infiltration. The activities of SOD and LDH and the content of GSH were reduced in the Xinshubao medium, high groups and the Di’ao Xinxuekang group while the content of MDA was significantly decreased(P<0.05,P<0.01). The levels of IL-18 and IL-1β in cardiac tissue were significantly decreased(P<0.05,P<0.01), and the expression of Bcl-2 was significantly up-regulated and Bax and Caspase 3 were significantly down-regulated(P<0.05,P<0.01). Conclusion Xinshubao has a protective effect on cardiac injury caused by ligation of descending branches before myocardial infarction, and this protective effect is related to the anti-inflammatory and anti-apoptotic effects.
作者
关雪
程美佳
王鑫淼
鞠业涛
王东妮
田苗
张会永
贾连群
杨关林
闵冬雨
GUAN Xue;CHENG Meijia;WANG Xinmiao;JU Yetao;WANG Dongni;TIAN Miao;ZHANG Huiyong;JIA Lianqun;YANG Guanlin;MIN Dongyu(School of Basic Medicine,Daqing Campus,Harbin Medical University,Daqing 166513,Heilongjiang,China;Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)
出处
《中华中医药学刊》
CAS
北大核心
2021年第9期112-117,I0019-I0021,共9页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金(81974548)
辽宁省科学技术厅自然基金指导项目(2019-ZD-0965)
辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金(zyzx1705)。
关键词
心舒宝
心肌梗死
氧化应激
炎症
凋亡
Xinshubao
myocardial infarction
oxidative stress
inflammation
apoptosis
