作者：王永贤，贾敏，高景峰，霍涛，许立刚

单位：

1.河北省石家庄市人民医院导管室2.河北医科大学第二医院心内科3.河北省唐山市丰润区人民医院CT室4.河北省保定市第三中心医院心内科

Units：

1.Catheterization Room, Shijiazhuang People's Hospital, Shijiazhuang 050000, China 2.Department of Cardiology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, China 3.CT Room, Tangshan Fengrun People's Hospital, Tangshan 063000, China 4.Department of Cardiology, Baoding Third Central Hospital, Baoding 071000, China

关键词：

心肌再灌注损伤; 丹参多酚酸盐; 己糖激酶Ⅱ; 线粒体;

Keywords：

Myocardial reperfusion injury; Salvianolate; Hexokinase Ⅱ; Mitochondrial

CLC：

R 542.2

DOI：

10.12114/j.issn.1008-5971.2023.00.020

Funds：

河北省卫生健康委医学科学研究课题（20210865）

摘要：

目的 探讨丹参多酚酸盐对大鼠离体心脏心肌缺血再灌注损伤的保护作用及其机制。方法 本实验时间为2021年2—8月。选取50只SD雄性大鼠，采用简单随机抽样法分为假手术组、模型组、丹参多酚酸盐组、己糖激酶Ⅱ（HKⅡ）抑制剂组、HKⅡ抑制剂+丹参多酚酸盐组，每组10只。模型组、丹参多酚酸盐组、HKⅡ抑制剂组、HKⅡ抑制剂+丹参多酚酸盐组通过Langendorff离体心脏灌流技术建立离体心脏心肌缺血再灌注损伤模型，其中假手术组不关闭灌流液；模型组建模后给予含有0.9%氯化钠溶液（20 mg/kg）的灌流液灌流；丹参多酚酸盐组建模后给予含有丹参多酚酸盐（20 mg/kg）的灌流液灌流；HKⅡ抑制剂组建模后给予含有HKⅡ抑制剂（10 mg/kg）的灌流液灌流；HKⅡ抑制剂+丹参多酚酸盐组建模后给予含有丹参多酚酸盐（20 mg/kg）及HKⅡ抑制剂（10 mg/kg）的灌流液灌流。检测五组大鼠心脏组织细胞色素C含量、腺苷酸[三磷酸腺苷（ATP）、二磷酸腺苷（ADP）、单磷酸腺苷（AMP）]含量，线粒体通透性转换孔（mPTP）吸光度及线粒体中HKⅡ、Bax表达水平。结果 模型组心脏组织细胞色素C含量高于假手术组，丹参多酚酸盐组低于模型组，HKⅡ抑制剂组高于模型组，HKⅡ抑制剂+丹参多酚酸盐组高于丹参多酚酸盐组（P<0.05）。模型组心脏组织ATP、ADP含量低于假手术组，丹参多酚酸盐组心脏组织ATP、ADP含量高于模型组，HKⅡ抑制剂组心脏组织ATP含量低于模型组，HKⅡ抑制剂+丹参多酚酸盐组心脏组织ATP、ADP含量低于丹参多酚酸盐组（P<0.05）。模型组mPTP吸光度高于假手术组，丹参多酚酸盐组低于模型组，HKⅡ抑制剂组高于模型组，HKⅡ抑制剂+丹参多酚酸盐组高于丹参多酚酸盐组（P<0.05）。模型组线粒体中HKⅡ表达水平低于假手术组、Bax表达水平高于假手术组，丹参多酚酸盐组线粒体中HKⅡ表达水平高于模型组、Bax表达水平低于模型组，HKⅡ抑制剂组线粒体中HKⅡ表达水平低于模型组，HKⅡ抑制剂+丹参多酚酸盐组线粒体中HKⅡ表达水平低于丹参多酚酸盐组、Bax表达水平高于丹参多酚酸盐组（P<0.05）。结论 丹参多酚酸盐可降低大鼠离体心脏心肌缺血再灌注损伤模型心脏组织细胞色素C含量、mPTP吸光度、线粒体中Bax表达水平，升高心脏组织ATP、ADP含量及线粒体中HKⅡ表达水平，丹参多酚酸盐改善大鼠心肌缺血再灌注损伤线粒体能量供应障碍的机制可能与其激活线粒体HKⅡ信号通路相关。

Abstract：

Objective To investigate the protective effect and mechanism of salvianolate on myocardial ischemia reperfusion injury in isolated rat heart. Methods This experiment lasted from February to August 2021. Fifty SD male rats were randomly divided into sham operation group, model group, salvianolate group, hexokinase Ⅱ (HKⅡ) inhibitor group, HKⅡ inhibitor+salvianolate group, with 10 rats in each group. The model group, salvianolate group, HKⅡ inhibitor group, HKⅡ inhibitor+salvianolate group were used to establish myocardial ischemia-reperfusion injury model by Langendorff isolated heart perfusion technique. In the sham operation group, the perfusion fluid was not closed; the model group was given perfusion solution containing 0.9% sodium chloride solution (20 mg/kg) after modeling; the salvianolate group was given perfusion solution containing salvianolate (20 mg/kg) after modeling; the HKⅡ inhibitor group was given perfusion solution containing HKⅡ inhibitor (10 mg/kg) after modeling; the HKⅡ inhibitor+salvianolate group was given perfusion solution containing salvianolate (20 mg/kg) and HKⅡ inhibitor (10 mg/kg) after modeling. The cytochrome C content and adenylate [adenosine triphosphate (ATP) , adenosine diphosphate (ADP) , adenosine monophosphate (AMP) ] content in heart tissues, mitochondrial permeability transition pore (mPTP ) absorbance, HKⅡ and Bax expression levels in mitochondria of the five groups were detected. Results The cytochrome C content in heart tissues of the model group was higher than that of the sham operation group, cytochrome C content in heart tissues of the salvianolate group was lower than that of the model group, cytochrome C content in heart tissues of the HKⅡ inhibitor group was higher than that of the model group, cytochrome C content in heart tissues of the HKⅡ inhibitor+salvianolate group was higher than that of the pretreatment group (P < 0.05) . The ATP and ADP contents in heart tissues of the model group were lower than those of the sham operation group, ATP and ADP contents in heart tissues of the salvianolate group were higher than those of the model group, ATP contents in heart tissues of the HKⅡ inhibitor group was lower than that of the model group, ATP and ADP contents in heart tissues of the HKⅡ inhibitor+salvianolate group were lower than those of the pretreatment group (P < 0.05) . The mPTP absorbance of the model group was higher than that of the sham operation group, mPTP absorbance of the salvianolate group was lower than that of the model group, mPTP absorbance of the HKⅡ inhibitor group was higher than that of the model group, mPTP absorbance of the HKⅡ inhibitor+salvianolate group was higher than that of the pretreatment group (P < 0.05) . The HKⅡ expression level in mitochondrial of the model group was lower than that of the sham operation group, Bax expression level in mitochondrial of the model group was higher than that of the sham operation group, HKⅡ expression level in mitochondrial of the salvianolate group was higher than that of the model group, Bax expression level in mitochondrial of the salvianolate group was lower than that of the model group, HKⅡ expression level in mitochondrial of the HKⅡ inhibitor group was lower than that of the model group, HKⅡ expression level in mitochondrial of the HKⅡ inhibitor+salvianolate group was lower than that of the pretreatment group, Bax expression level in mitochondrial of the HK Ⅱ inhibitor+salvianolate group was higher than that of the pretreatment group (P < 0.05) . Conclusion Salvianolate can reduce the cytochrome C content in heart tissues, mPTP absorbance, and Bax expression level in mitochondrial, and increase the ATP and ADP contents in heart tissues, and HKⅡ expression level in mitochondrial of isolated rat heart with myocardial ischemia-reperfusion injury. The protective mechanism of salvianolate on myocardial ischemia-reperfusion injury in isolated rat heart may be related to its activation of mitochondrial HKⅡ signal pathway.

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