目的 探索哈巴苷对高脂饲料诱导的动脉粥样硬化（atherosclerosis，AS）小鼠脂代谢和骨代谢的影响及其可能的作用机制。方法 高脂饲料喂养载脂蛋白E敲除（apolipoprotein E knock-out，ApoE^−/−）小鼠8周构建AS模型，随机分为模型组、阿托伐他汀（2.6 mg/kg）组和哈巴苷（20 mg/kg）组，每组8只。另取8只C57BL/6N野生型小鼠作为对照组，给予普通饲料喂养。药物干预9周后，利用micro-CT检测小鼠骨微结构的变化；利用红外光谱检测骨材料构成；利用苏木素-伊红（hematoxylin-eosin，HE）染色观察小鼠股骨骨小梁的形态，抗酒石酸酸性磷酸酶（tartrate resistant acid phosphatase，TRAP）染色观察小鼠股骨破骨细胞的数量；利用生化法检测小鼠血清总胆固醇（total cholesterol，TC）、高密度脂蛋白胆固醇（high density lipoprotein cholesterol，HDL-C）、低密度脂蛋白胆固醇（low density lipoprotein cholesterol，LDL-C）、TRAP、1型胶原交联C端末端肽（C-terminal telopeptide of type I collagen，CTX-1）水平；利用Western blotting检测骨组织活化T细胞核因子c1（nuclear factor-activated T cell 1，NFATc1）、原癌基因Fos（fos proto-oncogene，c-Fos）、组织蛋白酶K（cathepsin K）、过氧化物酶体增殖物激活受体γ共激活因子-1β（peroxisome proliferator-activated receptor γ coactivator-1β，PGC-1β）、雌激素相关受体α（estrogen-related receptor α，ERRα）和过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor γ，PPARγ）的蛋白表达。结果 哈巴苷显著改善AS小鼠骨微结构和骨材料构成（P＜0.05、0.01），降低血清中脂代谢TC、LDL-C和HDL-C水平以及AS指数（P＜0.05、0.01），降低血清中骨吸收特异性指标CTX-1和TRAP的水平（P＜0.01），降低骨组织中骨吸收相关蛋白c-Fos、NFATc1和cathepsin K的表达水平（P＜0.01）。此外，哈巴苷能抑制AS小鼠骨组织中PGC-1β、ERRα和PPARγ蛋白的表达（P＜0.05、0.01）。结论 哈巴苷可以改善AS小鼠的血脂代谢，抑制骨吸收，从而发挥改善骨质量作用。其作用机制可能与抑制PPARγ/ERRα/PGC-1β信号通路有关。

Objective To explore the effect and potential mechanism of harpagide on lipid metabolism and bone metabolism in atherosclerosis (AS) mice fed a high-fat diet. Methods AS model was constructed by feeding apolipoprotein E knock-out (ApoE^-/-) mice with high-fat diet for eight weeks, mice were randomly divided into model group, atorvastatin (2.6 mg/kg) group, and harpagide (20 mg/kg) group, with eight mice in each group. Additionally, eight C57BL/6N wild-type mice were included as control group and fed a standard diet. After nine weeks of drug intervention, micro-CT was used to analyze changes in bone microarchitecture. Infrared spectroscopy was employed to examine bone material composition. Hematoxylin-eosin (HE) staining was applied to observe the morphology of femoral trabeculae, and tartrate resistant acid phosphatase (TRAP) staining was used to quantify osteoclast numbers in the femur. Levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), TRAP and C-terminal telopeptide of type I collagen (CTX-1) in serum were measured using biochemical assays. Western blotting was performed to detect protein expression levels of nuclear factor of activated T cells 1 (NFATc1), Fos proto-oncogene (c-Fos), cathepsin K, peroxisome proliferator-activated receptor gamma coactivator-1β (PGC-1β), estrogen-related receptor α (ERRα) and peroxisome proliferator-activated receptor γ (PPARγ) in bone tissue. Results Harpagide significantly improved bone microarchitecture and bone material composition in AS mice (P < 0.05, 0.01), reduced levels of TC, LDL-C, HDL-C in serum and AS index (P < 0.05, 0.01), decreased levels of bone resorption markers CTX-1 and TRAP in serum (P < 0.01), down-regulated the expressions of bone resorption-related proteins c-Fos, NFATc1 and cathepsin K in bone tissue (P < 0.01). Additionally, harpagide suppressed the protein expression levels of PGC-1β, ERRα and PPARγ in bone tissue of AS mice (P < 0.05, 0.01). Conclusion Harpagide ameliorates dyslipidemia and suppresses bone resorption in AS mice, thereby exerting anti-osteoporotic effects. The mechanism may involve the inhibition of PPARγ/ERRα/PGC-1β signaling pathway.

R285.5

国家自然科学基金资助项目（82261138556）；2024年国家中医药管理局中医药国际合作专项（XDZYJZC-002）