目的 探究黄芪甲苷对糖尿病心肌病（diabetic cardiomyopathy，DCM）小鼠的心肌保护作用及作用机制。方法 随机选取10只db/m小鼠作为对照组，选取30只db/db小鼠将其随机分为模型组、黄芪甲苷（40 mg/kg）组和恩格列净（10 mg/kg）组，每组10只；采用酶联免疫吸附分析（enzyme linked immunosorbent assay，ELISA）法检测各组小鼠血清中肌酸激酶同工酶（creatine kinase-MB，CK-MB）、糖化血白蛋白（glycated albumin，GA）、三酰甘油（triglycerides，TG）、低密度脂蛋白胆固醇（low density lipoprotein cholesterol，LDL-C）、高密度脂蛋白胆固醇（high density lipoprotein cholesterol，HDL-C）、超氧化物歧化酶（super oxide dismutase，SOD）和白细胞介素-1β（interleukin-1β，IL-1β）水平；苏木素-伊红（hematoxylin-eosin，HE）、Masson和天狼星红染色观察各组小鼠心肌组织病理改变，免疫组织化学法和Western blotting法检测小鼠心肌组织中IL-1β、IL-6、CXC趋化因子配体-10（C-X-C motif chemokine ligand 10，CXCL-10）和IL-17信号通路相关蛋白表达量，qRT-PCR法检测小鼠心肌组织中IL-1β、IL-6、IL-17和IL-17受体（interleukin-17 receptor，IL-17R）mRNA表达量。结果 与对照组比较，模型组小鼠血清中CK-MB活性、GA含量、TG、LDL-C、SOD和IL-1β水平显著升高（P＜0.05、0.01）；心肌组织病理损伤明显，心肌纤维染色不均程度加重，心肌组织中呈紫红色心肌组织减少，蓝染的胶原纤维增多，心肌组织中呈红染的胶原纤维增多；免疫组织化学检测结果显示，心肌组织中IL-1β、IL-6和CXCL-10蛋白表达显著升高（P＜0.05、0.01）；Western blotting结果显示，心肌组织中IL-1β、IL-17、IL-17R、肿瘤坏死因子受体相关因子6（tumor necrosis factor receptor associated factor 6，TRAF6）、丝裂原活化蛋白激酶（mitogen-activated protein kinase，MAPKs）、c-Jun、核因子-κB（nuclear factor-kappa B，NF-κB）p65蛋白表达量显著增加（P＜0.01）；qRT-PCR结果显示，心肌组织中IL-1β、IL-6、IL-17和IL-17R mRNA表达水平显著降低（P＜0.05、0.01）。与模型组比较，给予黄芪甲苷治疗后，DCM小鼠血清中CK-MB活性、GA含量、TG、LDL-C、SOD和IL-1β水平显著降低（P＜0.05、0.01），HDL-C水平显著升高（P＜0.05）；心肌组织病理损伤减轻，心肌纤维染色不均程度减轻，心肌组织中以紫红色心肌组织为主，蓝色胶原纤维减少，心肌组织中红色胶原纤维减少；免疫组织化学检测结果显示，心肌组织中IL-1β、IL-6和CXCL-10蛋白表达显著降低（P＜0.05）；Western blotting结果显示，心肌组织中IL-1β、IL-17、IL-17R、TRAF6、MAPKs、c-Jun、NF-κB p65蛋白表达水平显著降低（P＜0.05、0.01）；qRT-PCR结果显示，心肌组织中IL-1β、IL-6、IL-17和IL-17R mRNA表达水平降低（P＜0.01）。结论 黄芪甲苷可抑制炎性因子的表达、改善DCM小鼠心脏功能，作用机制可能与抑制IL-17信号通路有关。

Objective To investigate the cardioprotective effects of astragaloside Ⅳ, an active component of the traditional Chinese medicine Astragalus membranaceus, on the myocardium of diabetic cardiomyopathy (DCM) mice, and to elucidate its underlying mechanisms. Methods A total of ten db/m mice were randomly selected as the control group, and 30 db/db mice were randomly divided into model group, astragaloside (40 mg/kg) group and engaglipzin (10 mg/kg) group, with ten mice in each group.Enzyme linked immunosorbent assay (ELISA) was used to detect creatine kinase-MB (CK-MB), glycated albumin (GA), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), super oxide dismutase (SOD) and interleukin-1β (IL-1β) levels in serum of mice in each group. Hematoxylin-eosin (HE), Masson and Sirius red staining were used to observe pathological changes in myocardial tissue of mice in each group. Immunohistochemical and Western blotting methods were used to detect the expression levels of IL-1β, IL-6, CXC motif chemokine ligand 10 (CXCL-10) and IL-17 signaling pathway-related proteins in mouse myocardial tissue. The mRNA expression of IL-1β, IL-6, IL-17 and IL-17 receptor (IL-17R) in mice myocardial tissue was detected by qRT-PCR. Results Compared with control group, CK-MB activity, GA content, TG, LDL-C, SOD and IL-1β levels in serum of mice in model group were significantly increased (P < 0.05, 0.01). Pathological damage to myocardial tissue was obvious, uneven degree of myocardial fiber staining was increased, purplish red myocardial tissue was reduced, blue-dyed collagen fibers were increased, and red-dyed collagen fibers were increased. Immunohistochemical test results showed that the expression of IL-1β, IL-6 and CXCL-10 protein in myocardial tissue was significantly increased (P < 0.05, 0.01). Western blotting results showed that expression of IL-1β, IL-17, IL-17R, tumor necrosis factor receptor associated factor 6 (TRAF6), mitogen activated protein kinase (MAPKs), c-Jun and nuclear factor-kappa B (NF-κB) p65 proteins in myocardial tissue were significantly increased (P < 0.01). Results of qRT-PCR showed that the mRNA expression levels of IL-1β, IL-6, IL-17 and IL-17R in myocardial tissue were significantly decreased (P < 0.05, 0.01). Compared with model group, after treatment with astragaloside Ⅳ, CK-MB activity, GA content, TG, LDL-C, SOD and IL-1β levels in serum of DCM mice were significantly decreased (P < 0.05, 0.01), while HDL-C level was significantly increased (P < 0.05). Pathological damage of myocardial tissue was alleviated, the degree of uneven staining of myocardial fibers was alleviated, the purplish red myocardial tissue was dominant, the blue collagen fibers were decreased, and the red collagen fibers were decreased. Immunohistochemical results showed that the expressions of IL-1β, IL-6 and CXCL-10 in myocardial tissue were significantly decreased (P < 0.05). Results of Western blotting showed that the expression levels of IL-1β, IL-17, IL-17R, TRAF6, MAPKs, c-Jun, NF-κB p65 protein in myocardial tissue were significantly decreased (P < 0.05, 0.01). The mRNA expression levels of IL-1β, IL-6, IL-17 and IL-17R in myocardial tissue were decreased by qRT-PCR (P < 0.01). Conclusion Astragaloside IV inhibited inflammatory factor expression and improved cardiac function in DMC mice. This protective effect may be achieved by inhibiting IL-17 signaling pathway.

R285.5

国家自然科学基金面上项目(82274504);河北省重点研发计划项目(20377711D,192777104D);河北省自然科学基金项目(H2023206203);河北省中医药管理局科研计划项目(2024038);河北省医学科学研究课题计划(20230474)