目的 探讨芪参益气滴丸对糖尿病心肌缺血损伤小鼠血管内皮功能及血管新生的作用机制。方法 102只雄性C57BL/6J小鼠随机分为对照组、模型组、尼可地尔片（4.55 mg/kg）组及芪参益气滴丸低、高剂量（227.5、455.0 mg/kg）组和芪参益气滴丸（227.5 mg/kg）＋EX-527（10 mg/kg）组。第2周开始对照组以正常饲料饲养，其余各组以高脂饲料饲养，然后持续7 d ip链脲霉素（50 mg/kg），建立2型糖尿病模型，对照组ip等体积生理盐水。第4周模型组和各给药组小鼠连续5 d颈背部sc异丙肾上腺素盐酸盐（100 mg/kg），对照组sc等体积生理盐水。第5周开始给予药物干预14 d，取血清测定心肌肌钙蛋白I（cardiac troponin I，cTn I）、肌酸激酶MB同工酶（creatine kinase-MB，CK-MB）、一氧化氮（nitric oxide，NO）、内皮素-1（endothelin-1，ET-1）和血管性血友病因子（von willebrand factor，vWF）含量；病理染色检测心脏形态结构、胶原纤维比值、心肌凋亡数量和血管新生情况；Western blotting检测心脏组织沉默信息调节因子1（silence information regulator 1，Sirt1）、内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）、磷酸化苏氨酸495位点eNOS（phosphorylation of Thr495-eNOS，pT495-eNOS）、磷酸化丝氨酸1177位点eNOS（phosphorylation of Ser1177-eNOS，pS1177-eNOS）蛋白表达。免疫荧光双标染色检测心脏Sirt1与eNOS共定位。结果 与对照组比较，模型组小鼠血清cTnI水平及CK-MB活性均明显增加（P<0.01），ET-1、vWF和NO水平均显著减少（P<0.01）；心肌细胞排列紊乱，肌纤维缺失，肌间隙增大，心肌纤维化面积和心肌凋亡数目显著增加（P<0.01），血小板-内皮细胞黏附分子（platelet endothelial cell adhesion molecule-1，CD31）标记的新生血管数量明显减少（P<0.01），心脏Sirt1、eNOS表达降低（P<0.01），pT495-eNOS水平升高（P<0.01），Sirt1和eNOS共定位减少。芪参益气滴丸干预后均有显著的保护效应，加入EX-527能部分逆转芪参益气滴丸的作用。结论 芪参益气滴丸可通过上调Sirt1和eNOS水平，增加Sirt1与eNOS相互作用，提高NO生物利用度，保护内皮功能，改善血管新生，抑制心肌纤维化和心肌凋亡，从而促进糖尿病心肌缺血损伤修复。

Objective To explore the mechanism of Qishen Yiqi Droplet (芪参益气滴丸) on vascular endothelial function and angiogenesis in diabetes mice with myocardial ischemia injury. Methods A total of 102 male C57BL/6J mice were randomly divided into control group, model group, Nicorandil Tablet (尼可地尔片, 4.55 mg/kg) group, Qishen Yiqi Droplet low-, high-dose (227.5, 455.0 mg/kg) groups and Qishen Yiqi Droplet (227.5 mg/kg) + EX-527 (10 mg/kg) group. In the second week, the control group was fed with normal diet, and the other groups were fed with high-fat diet. In the third week, streptozotocin (50 mg/kg) was intraperitoneally injected for 7 d to establish the model of type 2 diabetes. The control group was injected with equal dose of normal saline. At the fourth week, the model group and each administration group mice were subcutaneously injected with isoproterenol hydrochloride (100 mg/kg) on the nape of the neck for five consecutive days, while the control group mice were subcutaneously injected with the same volume of normal saline. After 14 d of drug intervention in the fifth week, serum was taken to measure cardiac troponin I (cTn I), creatine kinase MB isoenzyme (CK-MB), nitric oxide (NO), endothelin 1 (ET-1) and von willebrand factor (vWF) contents; Cardiac morphology, collagen fiber ratio, myocardial apoptosis and angiogenesis were detected by pathological staining; Western blotting was used to detect the expressions of silence information regulator 1 (Sirt1), endothelial nitric oxide synthase (eNOS), phosphorylation of Thr495-eNOS (pT495-eNOS) and phosphorylation of Ser1177-eNOS (pS1177-eNOS) proteins in cardiac tissue. Immunofluorescence double staining was used to detect the co-localization of Sirt1 and eNOS in the heart. Results Compared with control group, cTnI level and CK-MB activity in serum of mice in model group were significantly increased (P < 0.01), while the levels of ET-1, vWF, and NO were significantly reduced (P < 0.01); Myocardial cells were disordered, muscle fibers were missing, muscle space was enlarged, myocardial fibrosis area and myocardial apoptosis number were significantly increased (P < 0.01), the number of new blood vessels labeled by platelet endothelial cell adhesion molecule-1 (CD31) was significantly reduced (P < 0.01), the expressions of Sirt1 and eNOS in heart were reduced (P < 0.01), level of pT495-eNOS was increased (P < 0.01), the co-localization of Sirt1 and eNOS was reduced. After intervention, Qishen Yiqi Droplet showed significant protective effects, and the addition of EX-527 could partially reverse the effect of Qishen Yiqi Dropping Pills. Conclusion Qishen Yiqi Droplet can promote the repair of myocardial ischemia injury in diabetes by up-regulating the levels of Sirt1 and eNOS, increasing the interaction between Sirt1 and eNOS, improving the bioavailability of NO, protecting endothelial function, improving angiogenesis, inhibiting myocardial fibrosis and myocardial apoptosis.

R285.5

国家自然科学基金青年科学基金项目（82104627）；中央高校基本科研业务费专项（2022-JYB-XJSJJ-065）