目的 基于非靶向代谢组学探讨血脂康胶囊干预ApoE^-/-小鼠冠状动脉微循环障碍（coronary microvascular dysfunction，CMD）的作用机制。方法 SPF级ApoE^-/-小鼠高脂喂养7周后，尾iv月桂酸钠复制CMD模型，随机分为模型组、阿托伐他汀（6 mg/kg）组及血脂康低、中、高剂量（90、180、360 mg/kg）组，另取12只C57BL/6J小鼠作为对照组。给予药物干预8周后，采用苏木素-伊红（hematoxylin-eosin，HE）、Carstairs、Heidenhain染色检测小鼠心肌组织与心肌微血管形态与病理学变化；超声心动分析评估心脏功能性损伤；油红O染色评估主动脉脂质蓄积程度；生化法、ELISA法、免疫组化及免疫荧光检测血脂、心肌损伤及内皮功能相关指标；非靶向代谢组学技术分析小鼠粪便代谢物的变化，并分析差异代谢物与显著富集代谢通路；Western blotting检测心肌组织沉默信息调节因子1（silent information regulator 1，SIRT1）、过氧化物酶体增殖物激活受体γ共激活因子-1α（peroxisome proliferator activated receptor γ coactivator-1α，PGC-1α）、p-p65和p65蛋白表达。结果 与模型组比较，血脂康胶囊可显著降低小鼠血清中三酰甘油、总胆固醇、低密度脂蛋白胆固醇水平以及心肌损伤和血管内皮损伤程度（P＜0.05、0.01），显著改善小鼠微血栓、心肌缺血和心功能下降程度（P＜0.01）。非靶向代谢组学鉴定出14种差异代谢物和15条富集通路（P＜0.05）。Western blotting结果显示，与模型组比较，血脂康胶囊可显著上调心肌组织SIRT1、PGC-1α的蛋白表达（P＜0.01），并下调p-p65/p65的蛋白表达（P＜0.01）。结论 血脂康胶囊可发挥调血脂、改善内皮功能障碍和炎症反应作用，从而有效改善CMD模型小鼠心功能并减轻心肌结构损伤，其作用机制可能与调控精氨酸-脯氨酸-α-酮戊二酸代谢轴和eNOS/SIRT1/PGC-1α信号通路有关。

Objective To investigate the mechanism of Xuezhikang Capsules (血脂康胶囊) in intervening coronary microvascular dysfunction (CMD) in ApoE^−/− mice based on non-targeted metabolomics. Methods After seven weeks of high-fat diet feeding, SPF-grade ApoE^−/− mice were injected with sodium laurate via tail vein to establish CMD model. The mice were then randomly divided into model group, atorvastatin (6 mg/kg) group, Xuezhikang low-, medium-, and high-dose (90, 180, 360 mg/kg) groups, another 12 C57BL/6J mice were selected as the control group. After eight weeks of drug intervention, the morphology and pathological changes of myocardial tissue and myocardial microvessels in mice were detected using hematoxylin-eosin (HE), Carstairs and Heidenhain staining. Cardiac functional injury was evaluated by echocardiography. Oil red O staining was used to evaluate the degree of lipid accumulation in the aorta. Lipid levels, myocardial injury and endothelial function-related indicators were detected by biochemical assays, ELISA, immunohistochemistry and immunofluorescence. The changes in fecal metabolites were analyzed using non-targeted metabolomics, and differential metabolites and significantly enriched metabolic pathways were identified. Western blotting was used to detect silent information regulator 1 (SIRT1), peroxisome proliferator activated receptor γ coactivator-1α (PGC-1α), p-p65 and p65 protein expressions in myocardial tissue. Results Compared with model group, Xuezhikang Capsules significantly reduced the levels of triglycerides, total cholesterol, low-density lipoprotein cholesterol, as well as the degree of myocardial and endothelial injury in serum of mice (P < 0.05, 0.01), and significantly improved the degree of microthrombus, myocardial ischemia and heart function decline in mice (P < 0.01). Non-targeted metabolomics identified 14 differentially expressed metabolites and 15 enriched pathways (P < 0.05). Western blotting results showed that compared with model group, Xuezhikang Capsules significantly up-regulated the protein expressions of SIRT1 and PGC-1α in myocardial tissue (P < 0.01), and down-regulated the protein expression of p-p65/p65 (P < 0.01). Conclusion Xuezhikang Capsules could regulate blood lipids, improve endothelial dysfunction and inflammatory response, thereby effectively improving cardiac function and reducing myocardial structural damage in CMD model mice. Its mechanism may be related to the regulation of arginine-proline-α-ketoglutarate metabolic axis and eNOS/SIRT1/PGC-1α signaling pathway.

R285.5

国家自然科学基金资助项目（81503287）；北京市自然科学基金资助项目（7232287）