目的 探討生白術(shù)發(fā)揮健脾作用的有效部位及化學(xué)成分。方法 采用“ig番瀉葉＋疲勞游泳＋饑飽失常”的方法復(fù)制脾虛大鼠模型，給予生白術(shù)水提物、多糖及除糖部位治療，通過一般行為學(xué)特征、小腸推進率及胃內(nèi)殘留率、消化吸收功能、水液代謝功能、組織病理結(jié)構(gòu)和神經(jīng)遞質(zhì)等指標(biāo)，評價生白術(shù)治療脾虛的有效部位；進一步采用HP-20型大孔樹脂柱色譜制備除糖部位不同體積分?jǐn)?shù)乙醇洗脫組分，通過α-二氟甲基鳥氨酸（difluoromethylornithine，DFMO）誘導(dǎo)的大鼠小腸上皮細胞（intestinal epithelial cell-6，IEC-6）生長抑制模型，評價不同濃度乙醇洗脫組分對細胞活力的影響；最后采用超高效液相色譜-串聯(lián)四極桿飛行時間質(zhì)譜（UPLC-Q-TOF/MS）技術(shù)對有效組分進行化學(xué)成分表征，并定向分離主要化學(xué)成分，評估其對DFMO誘導(dǎo)的IEC-6細胞活力的影響。結(jié)果 生白術(shù)水提物及其除糖部位主要通過調(diào)節(jié)神經(jīng)遞質(zhì)水平、促進胃腸動力、修復(fù)組織損傷和提高消化吸收能力、炎癥因子水平顯著改善脾虛大鼠的癥狀；除糖部位中90%乙醇洗脫組分（CT-90E）在0.1～0.15 mg/L可逆轉(zhuǎn)DFMO所致的IEC-6細胞生長抑制；CT-90E組分中的8-表白術(shù)內(nèi)酯（12.5～100 μmol/L）、白術(shù)內(nèi)酯Ⅱ（50～100 μmol/L）、白術(shù)內(nèi)酯Ⅰ（25～50 μmol/L）及鄰苯二甲酸（12.5～50 μmol/L）可逆轉(zhuǎn)DFMO誘導(dǎo)的IEC-6細胞生長抑制。結(jié)論 生白術(shù)水提物治療脾虛藥效顯著，除糖部位是其治療脾虛的活性部位，其中CT-90E組分可促進腸黏膜的修復(fù)，而8-表白術(shù)內(nèi)酯、白術(shù)內(nèi)酯Ⅱ、白術(shù)內(nèi)酯Ⅰ及鄰苯二甲酸為CT-90E組分中修復(fù)腸黏膜的主要活性成分。

Objective To explore the active fractions and chemical components with spleen-invigorating effect from raw rhizomes of Atractylodes macrocephala. Methods The rat model of spleen-deficiency was replicated by the method of “ig senna leaves + fatigued swimming + abnormal hunger and satiety”, Which was treated with water extract from raw rhizomes of A. macrocephala, polysaccharides and non-polysaccharide parts. The effective fractions of raw rhizomes of A. macrocephala in treating spleen deficiency were evaluated using various indicators, including general behavioral characteristics, gastrointestinal (GI) transit rate, gastric residual rate, digestive and absorption functions, water and fluid metabolism, tissue pathological structure, and neurotransmitters of rats with spleen-deficiency. The non-polysaccharide part was subjected to chromatography on a HP-20 macroporous resin column eluted with different concentrations of aqueous ethanol. The intestine epithelial cell (IEC-6) growth inhibition model induced by α-difluoromethylornithine (DFMO) was applied to assay the cell viability of the obtained fractions. The chemical constituents in the active fraction were analyzed by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The main constituents were isolated and evaluated for their impact on IEC-6 cell viability induced by DFMO. Results The water extract and non-polysaccharide part from raw rhizomes of A. macrocephala significantly improved the symptoms of spleen-deficiency rats, mainly by regulating neurotransmitter levels, promoting gastrointestinal motility, repairing tissue damage, enhancing digestive and absorption capabilities, and modulating inflammatory factors. The 90% ethanol-elution fraction from non-polysaccharide part (CT-90E) at concentrations of 0.1—0.15 mg/L could reverse the growth inhibition of IEC-6 cells induced by DFMO. The main components in CT-90E, including 8-epiatylenolide (12.5—100 μmol/L), atractylenolide II (50—100 μmol/L), atractylenolide I (25—50 μmol/L), and phthalic acid (12.5—50 μmol/L) could reverse the growth inhibition of IEC-6 cells induced by DFMO. Conclusion The water extract of raw rhizomes of A. macrocephala exhibits significant effect in treating spleen deficiency, with the non-polysaccharide part identified as the active part responsible for treating spleen deficiency. Among the fractions of non-polysaccharide part, CT-90E promotes the repair of intestinal mucosa, with 8-epiatylenolide, atractylenolide II, atractylenolide I, and phthalic acid identified as the main active components responsible for mucosal repair.

R284.1;R285

國家自然科學(xué)基金資助項目（82073992）；中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)與健康科技創(chuàng)新工程項目（2023-I2M-2-006）