目的 探讨栀子苷对冠心病模型大鼠巨噬细胞极化及GATA结合蛋白1（GATA1）/β2肾上腺素受体（β2AR）信号通路的影响。方法 采用结扎左心室回旋分支末端的方法制备冠心病模型，模型大鼠分为模型组，栀子苷低、高剂量（25、100 mg·kg^-1）组，栀子苷（100 mg·kg^-1）＋克伦特罗（β2AR激动剂，0.5 mg·kg^-1）组，另设置假手术组，操作相同但不结扎。造模成功1周后开始给药，每天1次，栀子苷ig给药，克伦特罗ip给药，假手术组和模型组均ig等量的0.9%氯化钠溶液，连续给药3周。超声心动图进行大鼠心功能检测；ELISA检测血清N端B型利钠肽原（NT-proBNP）、白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α、IL-10及精氨酸酶-1（Arg-1）水平；流式细胞术检测心脏巨噬细胞相对含量及M1/M2极化状态；免疫组化法进行心肌组织内皮型一氧化氮合酶（eNOS）、内皮素1（ET-1）蛋白表达水平检测；采用Masson和TTC染色分别评估大鼠心肌病理形态学变化及心肌梗死情况；TUNEL染色检测心肌细胞凋亡情况；Western blotting法检测心肌组织Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤-2蛋白（Bcl-2）、裂解型半胱天冬酶-3（cleaved Caspase-3）、GATA1、β2AR蛋白表达水平。结果 模型组心肌组织相较于假手术组呈大量蓝色胶原纤维沉积，左室舒张末期容积（LVEDV）、左室收缩末期容积（LVESV）、血清NT-proBNP、TNF-α、IL-6含量、心脏组织中巨噬细胞数量、M1-F4/80⁺CD86⁺值（M1型巨噬细胞占比）、ET-1、心肌梗死率、细胞凋亡率、Bax、cleaved Caspase-3、β2AR表达水平显著升高（P＜0.05、0.01、0.001），而左室射血分数（LVEF）、左室缩短分数（LVFS）、M2-F4/80⁺CD163⁺值（M2型巨噬细胞占比）、eNOS、GATA1、Bcl-2表达水平显著降低（P＜0.01）；栀子苷低、高剂量给药可显著逆转模型组上述指标的变化趋势（P＜0.05、0.01），且显著升高IL-10、Arg-1水平（P＜0.05、0.01）；给予克伦特罗则明显抵消高剂量栀子苷对模型组大鼠上述指标的改善作用（P＜0.05）。结论 冠心病模型大鼠存在M1/M2型巨噬细胞极化失衡，以M1型巨噬细胞为主；栀子苷可通过调控GATA1/β2AR信号轴，促进巨噬细胞向M2极化，抑制M1型巨噬细胞活化，改善冠心病大鼠心肌损伤。

Objective The effects of geniposide on macrophage polarization and the GATA binding protein 1 (GATA1)/β2 adrenergic receptor (β2AR) signaling pathway in a rat model of coronary heart disease were investigated.Methods A rat model of coronary heart disease was established by ligating the end of the left circumflex branch of the left ventricle. The successfully modeled rats were divided into the model group, low-dose (25 mg·kg^-1) and high-dose (100 mg·kg^-1) geniposide groups, and the high-dose geniposide (100 mg·kg^-1) + clenbuterol (β2AR agonist, 0.5 mg·kg^-1) group. A sham operation group was also set up, with the same operation but without ligation. Drug administration began one week after successful modeling, once a day. Geniposide was ig administered, and clenbuterol was ip administered. The sham operation group and the model group were ig given the same volume of 0.9% sodium chloride solution. The treatment lasted for three weeks. Echocardiography was used to detect cardiac function in rats; ELISA was used to detect the levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-10, and arginase-1 (Arg-1) in serum; flow cytometry was used to detect the relative content of cardiac macrophages and M1/M2 polarization status; immunohistochemistry was used to detect the expression levels of endothelial nitric oxide synthase (eNOS) and endothelin-1 (ET-1) in myocardial tissue; Masson and TTC staining were used to evaluate the pathological morphology of myocardial tissue and myocardial infarction in rats; TUNEL staining was used to detect myocardial cell apoptosis; Western blotting was used to detect the expression levels of Bcl-2 associated X protein (Bax), B-cell lymphoma-2 protein (Bcl-2), cleaved caspase-3, GATA1, and β2AR in myocardial tissue.Results Compared with the sham operation group, the myocardial tissue of the model group showed a large amount of blue collagen fiber deposition. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), serum NT-proBNP, TNF-α, IL-6 content, the number of macrophages in cardiac tissue, M1-F4/80+CD86+ value (proportion of M1-type macrophages), ET-1, myocardial infarction rate, apoptosis rate, Bax, cleaved Caspase-3, and β2AR expression levels were significantly increased (P<0.05, 0.01, 0.001), while the left ventricular ejection fraction (LVEF), left ventricular fractional shortening (LVFS), M2-F4/80⁺CD163⁺ value (proportion of M2-type macrophages), eNOS, GATA1, and Bcl-2 expression levels were significantly decreased (P<0.01). Low-dose and high-dose geniposide administration could significantly reverse the changes in the above indicators in the model group (P<0.05, 0.01), and significantly increase the levels of IL-10 and Arg-1 (P<0.05, 0.01). Administration of clenbuterol significantly canceled the improvement effect of high-dose geniposide on the above indicators in the model group (P<0.05).Conclusion In the rat model of coronary heart disease, there is polarization imbalance of M1/M2 type macrophages, with M1 type macrophages being dominant. Gardenia glycoside can promote the polarization of macrophages towards M2 and inhibit the activation of M1-type macrophages by regulating the GATA1/β2AR signaling axis, thereby improving myocardial injury in rats with coronary heart disease.

R285.5

河南省医学科技攻关项目计划（20203A519）