目的 探究原花青素B2（procyanidins B2，PCB2）治疗肥胖型多囊卵巢综合征（polycystic ovary syndrome，PCOS）的作用机制。方法 建立肥胖型PCOS大鼠模型，使用不同剂量的PCB2治疗肥胖型PCOS大鼠。苏木素-伊红（hematoxylin-eosin，HE）染色观察卵巢组织形态；酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）法检测激素水平、乳酸、丙酮酸水平以及腺嘌呤核苷三磷酸（adenosine triphosphate，ATP）含量；免疫组化法检测增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）、半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、肝激酶B1（liver kinase B1，LKB1）和腺苷酸活化蛋白激酶（adenosine 5'-monophosphate-activated protein kinase，AMPK）的蛋白表达；蛋白质免疫印迹（Western blotting，WB）检测凋亡标志物、糖酵解关键限速酶以及LKB1/AMPK通路相关蛋白的表达。结果 与对照组比较，模型组卵泡闭锁，囊肿性卵泡数量增加，颗粒细胞层减薄，黄体减少；卵泡刺激素（follicle-stimulating hormone，FSH）、雌二醇（estradiol，‌‌E2）降低（P＜0.01），黄体生成素（luteinising hormone，LH）、睾酮（testosterone，T）、LH/FSH值升高（P＜0.01）；乳酸（lactic acid，LD）、ATP含量降低（P＜0.001）、丙酮酸升高（P＜0.001）；PCNA、AMPK、LKB1表达下调（P＜0.01），Caspase-3表达上调（P＜0.01）；B淋巴细胞瘤-2（B-cell lymphoma-2，‌Bcl-2）蛋白表达降低（P＜0.01）；Bcl-2相关X蛋白（Bcl-2-associated X protein，Bax）蛋白表达增加（P＜0.01），糖酵解关键限速酶乳酸脱氢酶A（lactate dehydrogenase A，LDHA）和肌肉丙酮酸激酶同工酶2（pyruvate kinase isozyme type M2，PKM2）以及LKB1、AMPKα、磷酸化-AMPKα（phosphorylated-AMPKα，p-AMPKα）蛋白表达显著降低（P＜0.01）。与模型组比较，各剂量PCB2均逆转了以上变化（P＜0.05、0.01、0.001），缓解了PCOS。结论 PCB2可能通过LKB1/AMPK轴调控糖酵解代谢途径治疗肥胖型PCOS。

Objective To investigate the specific mechanism of procyanidins B2 (PCB2) in treating obesity-related polycystic ovary syndrome (PCOS). Methods An obesity-induced PCOS rat model was established and treated with different doses of PCB2. Hematoxylin-eosin (HE) staining was used to observe the morphology of the ovarian tissue, enzyme linked immunosorbent assay (ELISA) was used to detect hormone level, actic acid and pyruvic acid levels and adenosine triphosphate (ATP) content, immunohistochemistry (IHC) was used to detect proliferating cell nuclear antigen (PCNA), cystein-asparate protease-3 (Caspase-3), liver kinase B1 (LKB1) and adenosine 5'-monophosphate-activated protein kinase (AMPK) protein expression and Western blotting (WB) was used to detect apoptotic markers, key rate-limiting enzymes of glucose metabolism, and LKB1/AMPK pathway-related proteins.Results Compared with the control group, the model group showed increased follicular atresia, increased cystic follicles, thinner granular cell layers, and fewer corpora lutea, decreased follicle-stimulating hormone (FSH) and estradiol (E2) (P < 0.01), increased luteinising hormone (LH), testosterone (T), and increased LH/FSH (P < 0.01), decreased lactic acid (LD) and adenosine triphosphate (ATP) (P<0.001), increased pyruvic acid (P < 0.001), downregulated PCNA, AMPK, and LKB1 expression (P < 0.01), and upregulated Caspase-3 expression (P < 0.01), decreased B-cell lymphoma-2 (Bcl-2) protein expression (P < 0.01) and increased Bcl-2-associated X protein(Bax) expression (P < 0.01), significantly decreased lactate dehydrogenase (LDHA) and pyruvate kinase isozyme type M2 (PKM2), and LKB1, AMPKα, and phosphorylated-AMPKα (p-AMPKα) protein expression (P < 0.01). Compared with the model group, all doses of PCB2 reversed these changes and alleviated the severity of PCOS (P < 0.05, 0.01, 0.001). Conclusion PCB2 may treat obesity-related PCOS by regulating the glycolytic metabolic pathway through the LKB1/AMPK axis.

R285.5

2022年重庆市科研机构绩效激励引导专项项目（jxyn2022-6）;重庆市科卫联合中医药科研项目（2023MXSM161）;中医“肾”藏象理论诊疗实践多学科交叉创新团队[渝中医（2022）33号];传承创新中心学科分层分类建设-中医妇科学（培育）[（2024）48号];第七批全国老中医药专家学术经验继承项目[国中医药人教函（2022）76号]