目的 探討梔子苷對冠心病模型大鼠巨噬細胞極化及GATA結(jié)合蛋白1（GATA1）/β2腎上腺素受體（β2AR）信號通路的影響。方法 采用結(jié)扎左心室回旋分支末端的方法制備冠心病模型，模型大鼠分為模型組，梔子苷低、高劑量（25、100 mg·kg^-1）組，梔子苷（100 mg·kg^-1）＋克倫特羅（β2AR激動劑，0.5 mg·kg^-1）組，另設置假手術(shù)組，操作相同但不結(jié)扎。造模成功1周后開始給藥，每天1次，梔子苷ig給藥，克倫特羅ip給藥，假手術(shù)組和模型組均ig等量的0.9%氯化鈉溶液，連續(xù)給藥3周。超聲心動圖進行大鼠心功能檢測；ELISA檢測血清N端B型利鈉肽原（NT-proBNP）、白細胞介素（IL）-6、腫瘤壞死因子（TNF）-α、IL-10及精氨酸酶-1（Arg-1）水平；流式細胞術(shù)檢測心臟巨噬細胞相對含量及M1/M2極化狀態(tài)；免疫組化法進行心肌組織內(nèi)皮型一氧化氮合酶（eNOS）、內(nèi)皮素1（ET-1）蛋白表達水平檢測；采用Masson和TTC染色分別評估大鼠心肌病理形態(tài)學變化及心肌梗死情況；TUNEL染色檢測心肌細胞凋亡情況；Western blotting法檢測心肌組織Bcl-2相關(guān)X蛋白（Bax）、B細胞淋巴瘤-2蛋白（Bcl-2）、裂解型半胱天冬酶-3（cleaved Caspase-3）、GATA1、β2AR蛋白表達水平。結(jié)果 模型組心肌組織相較于假手術(shù)組呈大量藍色膠原纖維沉積，左室舒張末期容積（LVEDV）、左室收縮末期容積（LVESV）、血清NT-proBNP、TNF-α、IL-6含量、心臟組織中巨噬細胞數(shù)量、M1-F4/80⁺CD86⁺值（M1型巨噬細胞占比）、ET-1、心肌梗死率、細胞凋亡率、Bax、cleaved Caspase-3、β2AR表達水平顯著升高（P＜0.05、0.01、0.001），而左室射血分數(shù)（LVEF）、左室縮短分數(shù)（LVFS）、M2-F4/80⁺CD163⁺值（M2型巨噬細胞占比）、eNOS、GATA1、Bcl-2表達水平顯著降低（P＜0.01）；梔子苷低、高劑量給藥可顯著逆轉(zhuǎn)模型組上述指標的變化趨勢（P＜0.05、0.01），且顯著升高IL-10、Arg-1水平（P＜0.05、0.01）；給予克倫特羅則明顯抵消高劑量梔子苷對模型組大鼠上述指標的改善作用（P＜0.05）。結(jié)論 冠心病模型大鼠存在M1/M2型巨噬細胞極化失衡，以M1型巨噬細胞為主；梔子苷可通過調(diào)控GATA1/β2AR信號軸，促進巨噬細胞向M2極化，抑制M1型巨噬細胞活化，改善冠心病大鼠心肌損傷。

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

河南省醫(yī)學科技攻關(guān)項目計劃（20203A519）