目的 探究原花青素B2（procyanidins B2，PCB2）治療肥胖型多囊卵巢綜合征（polycystic ovary syndrome，PCOS）的作用機制。方法 建立肥胖型PCOS大鼠模型，使用不同劑量的PCB2治療肥胖型PCOS大鼠。蘇木素-伊紅（hematoxylin-eosin，HE）染色觀察卵巢組織形態(tài)；酶聯(lián)免疫吸附試驗（enzyme linked immunosorbent assay，ELISA）法檢測激素水平、乳酸、丙酮酸水平以及腺嘌呤核苷三磷酸（adenosine triphosphate，ATP）含量；免疫組化法檢測增殖細(xì)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）的蛋白表達(dá)；蛋白質(zhì)免疫印跡（Western blotting，WB）檢測凋亡標(biāo)志物、糖酵解關(guān)鍵限速酶以及LKB1/AMPK通路相關(guān)蛋白的表達(dá)。結(jié)果 與對照組比較，模型組卵泡閉鎖，囊腫性卵泡數(shù)量增加，顆粒細(xì)胞層減薄，黃體減少；卵泡刺激素（follicle-stimulating hormone，F(xiàn)SH）、雌二醇（estradiol，‌‌E2）降低（P＜0.01），黃體生成素（luteinising hormone，LH）、睪酮（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表達(dá)下調(diào)（P＜0.01），Caspase-3表達(dá)上調(diào)（P＜0.01）；B淋巴細(xì)胞瘤-2（B-cell lymphoma-2，‌Bcl-2）蛋白表達(dá)降低（P＜0.01）；Bcl-2相關(guān)X蛋白（Bcl-2-associated X protein，Bax）蛋白表達(dá)增加（P＜0.01），糖酵解關(guān)鍵限速酶乳酸脫氫酶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）蛋白表達(dá)顯著降低（P＜0.01）。與模型組比較，各劑量PCB2均逆轉(zhuǎn)了以上變化（P＜0.05、0.01、0.001），緩解了PCOS。結(jié)論 PCB2可能通過LKB1/AMPK軸調(diào)控糖酵解代謝途徑治療肥胖型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年重慶市科研機構(gòu)績效激勵引導(dǎo)專項項目（jxyn2022-6）;重慶市科衛(wèi)聯(lián)合中醫(yī)藥科研項目（2023MXSM161）;中醫(yī)“腎”藏象理論診療實踐多學(xué)科交叉創(chuàng)新團隊[渝中醫(yī)（2022）33號];傳承創(chuàng)新中心學(xué)科分層分類建設(shè)-中醫(yī)婦科學(xué)（培育）[（2024）48號];第七批全國老中醫(yī)藥專家學(xué)術(shù)經(jīng)驗繼承項目[國中醫(yī)藥人教函（2022）76號]