目的 系統(tǒng)解析分級(jí)醇沉玉竹多糖（Polygonatum odoratum polysaccharides，POPs）各組分的精細(xì)結(jié)構(gòu)，通過(guò)網(wǎng)絡(luò)藥理學(xué)預(yù)測(cè)其治療糖尿病的作用機(jī)制，并對(duì)其抗氧化與α-葡萄糖苷酶抑制活性進(jìn)行評(píng)價(jià)。方法 采用梯度調(diào)節(jié)乙醇終體積分?jǐn)?shù)（35%、55%、70%、85%）分離獲得4種POPs組分（POP-35、POP-55、POP-70、POP-85），通過(guò)傅里葉變換紅外光譜（fourier transform infrared spectroscopy，F(xiàn)T-IR）、核磁共振波譜（nuclear magnetic resonance，NMR）、氣相色譜-質(zhì)譜聯(lián)用（gas chromatography-mass spectrometry，GC-MS）、高效凝膠滲透色譜（high performance gel permeation chromatography，HPGPC）和高效陰離子交換色譜（high performance anion exchange chromatography，HPAEC）聯(lián)合表征其理化性質(zhì)，解析分子結(jié)構(gòu)；基于網(wǎng)絡(luò)藥理學(xué)和分子對(duì)接技術(shù)，預(yù)測(cè)POPs治療糖尿病的作用機(jī)制；采用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picryl-hydrazyl，DPPH）自由基清除法、2,2'-聯(lián)氮-雙-3-乙基苯并噻唑啉-6-磺酸 [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)，ABTS]自由基清除法、Fe³⁺還原力測(cè)定法和α-葡萄糖苷酶抑制試驗(yàn)對(duì)POPs抗糖尿病活性進(jìn)行評(píng)價(jià)；結(jié)果 4種POPs均為低相對(duì)分子質(zhì)量（1.8×10³～2.4×10³）中性雜多糖，由果糖（84%～86%）與葡萄糖（14%～16%）構(gòu)成。其中均一多糖POP-55的結(jié)構(gòu)特征通過(guò)GC-MS與NMR波譜得以明確，其主鏈β-(2→1)-果糖殘基和α-(1→6)-葡萄糖殘基構(gòu)成，側(cè)鏈含β-(2→6)-果糖殘基構(gòu)成分支。網(wǎng)絡(luò)藥理學(xué)分析發(fā)現(xiàn)POP-55抗糖尿病的相關(guān)基因顯著富集于碳水化合物消化吸收、胰島素抵抗等信號(hào)通路，在相關(guān)基因中通過(guò)拓?fù)鋵W(xué)分析篩選出信號(hào)轉(zhuǎn)導(dǎo)與轉(zhuǎn)錄激活因子3（signal transducer and activator of transcription 3，STAT3）、半胱氨酸天冬氨酸蛋白酶3（Caspase 3，CASP3）、白細(xì)胞介素-2（interleukin-2，IL-2）等6個(gè)核心基因，分子對(duì)接證實(shí)其都與POP-55有較好的結(jié)合能力。體外實(shí)驗(yàn)表明，分級(jí)醇沉POPs具有顯著的抗氧化和α-葡萄糖苷酶抑制活性，并且具有濃度相關(guān)性。結(jié)論 分級(jí)醇沉POPs為菊粉型果聚糖，其降糖效應(yīng)可能通過(guò)競(jìng)爭(zhēng)性抑制α-葡萄糖苷酶活性延緩碳水化合物分解和調(diào)控IL-2介導(dǎo)的炎癥信號(hào)減輕胰島β細(xì)胞氧化損傷雙重途徑實(shí)現(xiàn)。

Objective This study systematically elucidates the fine structural characteristics of graded ethanol-precipitated Polygonatum odoratum polysaccharides (POPs), predicts their therapeutic mechanisms against diabetes through network pharmacology, and evaluates their antioxidant and α-glucosidase inhibitory activities. Methods Four POPs fractions (POP-35, POP-55, POP-70, POP-85) were isolated by gradient ethanol precipitation (35%, 55%, 70%, 85%). Their physicochemical properties and molecular structures were characterized using Fourier-transform infrared spectroscopy (FT-IR), gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), high-performance gel permeation chromatography (HPGPC), and high-performance anion-exchange chromatography (HPAEC). Network pharmacology and molecular docking were employed to predict antidiabetic mechanisms. Antidiabetic activities were assessed via DPPH/ABTS radical scavenging assays, ferric ion reducing power assay, and α-glucosidase inhibition assay. Results All four POPs fractions were low-molecular-weight (1.8×10³—2.4×10³) neutral heteropolysaccharides composed of fructose (84%—86%) and glucose (14%—16%). The homogeneous polysaccharide POP-55 exhibited a backbone of →2)-β-D-Fruf-(1→ glycosidic linkages with minor α-D-Glcp-(1→6) branched side chains, as confirmed by GC-MS and NMR. Network pharmacology analysis revealed that POP-55-related antidiabetic genes were significantly enriched in pathways such as carbohydrate digestion and absorption, insulin resistance and so on. Topological analysis identified six core targets (STAT3, CASP3, IL2, etc.) among these genes, which were further validated by molecular docking to exhibit strong binding affinities with POP-55. Conclusion Graded ethanol-precipitated POPs are inulin-type fructans exerting hypoglycemic effects through dual mechanisms: competitively inhibiting α-glucosidase to delay carbohydrate hydrolysis, and regulating IL2-mediated inflammatory signaling to alleviate oxidative damage in pancreatic β-cells.

R284.1

國(guó)家自然科學(xué)基金青年項(xiàng)目（82304133）；江蘇省自然科學(xué)基金青年項(xiàng)目（BK20230590）