目的 以油酸誘導(dǎo)脂代謝紊亂建立肝胰島素抵抗（IR）細(xì)胞模型，研究中藥單體成分小檗堿、黃芩苷、葛根素和甘草苷的體外降糖作用，為中藥降糖組方或成分配伍優(yōu)化提供研究基礎(chǔ)。方法 采用油酸不同濃度（0.1、0.2、0.5、1.0 mmol/L）及不同作用時(shí)間點(diǎn)（24、36、48 h）誘導(dǎo)HepG2細(xì)胞，建立IR-HepG2細(xì)胞模型，葡萄糖氧化酶法測(cè)定細(xì)胞葡萄糖消耗量，CCK-8法檢測(cè)細(xì)胞活力，確立模型的油酸最佳誘導(dǎo)濃度及最佳作用時(shí)間；蒽酮法和三酰甘油氧化酶法檢測(cè)IR模型細(xì)胞內(nèi)肝糖原和三酰甘油水平；油紅O染色檢測(cè)細(xì)胞形態(tài)的變化；觀察葡萄糖消耗量檢測(cè)IR模型的穩(wěn)定性。采用油酸最佳誘導(dǎo)濃度及最佳作用時(shí)間建立IR-HepG2細(xì)胞模型，研究不同劑量小檗堿、葛根素、黃芩苷和甘草苷對(duì)細(xì)胞葡萄糖消耗量、糖原含量、三酰甘油及細(xì)胞活力的影響。結(jié)果 油酸誘導(dǎo)IR-HepG2細(xì)胞模型最佳濃度1 mmol/L，最佳作用時(shí)間24 h，此時(shí)與對(duì)照組比較，HepG2細(xì)胞肝糖原含量明顯下降（P<0.001），三酰甘油顯著上升（P<0.01），模型建立后可至少持續(xù)穩(wěn)定36 h以上。與IR模型組比較，給藥干預(yù)24 h后，不同濃度小檗堿（5、10、20、50 μmol/L）、黃芩苷（1、5、10、20、50 μmol/L）、葛根素（20、40、80、160 μmol/L）和1 μmol/L甘草苷顯著增加葡萄糖消耗量（P<0.05、0.01、0.001）。與IR組比較，不同濃度小檗堿（10、20、50 μmol/L）、黃芩苷（20、50 μmol/L）、葛根素（10、20、80、160 μmol/L）顯著提高肝糖原含量（P<0.001）；甘草苷升高糖原含量但是差異不顯著。與對(duì)照組比較，除160 μmol/L葛根素和1 μmol/L黃芩苷顯著抑制細(xì)胞活力（P<0.05）外，其他組對(duì)細(xì)胞活力的影響均不顯著，結(jié)論 1 mmol/L油酸誘導(dǎo)24 h后能夠建立穩(wěn)定IR-HepG2細(xì)胞模型，適合作為高脂飲食誘導(dǎo)2型糖尿病的體外肝IR細(xì)胞模型。小檗堿、黃芩苷和葛根素顯著增加油酸誘導(dǎo)的IR-HepG2細(xì)胞葡萄糖消耗量和糖原合成，進(jìn)而改善脂代謝誘發(fā)的肝IR。

Objective To investigate the anti-hyperglycemic effect of the Chinese medicine ingredients such as berberine, baicalin, puerarin and liquiritin on the optimal insulin resistance (IR)-HepG2 cell model by oleic acid. It would provide the theoretical basis for the optimization of Chinese medicine prescription or anti-hyperglycemic components combination. Methods Different concentrations (0.1, 0.2, 0.5, and 1.0 mmol/L) of oleic acid were used to induce HepG2 cells for different time (24, 36 and 48 h), the glucose consumption was measured by glucose oxidase assay, and cell viability was detected by CCK-8 assay to define the optimal inducing concentration and time for IR-HepG2 cell model. Then the cell morphological changes were detected by oil red O staining. Finally, the stability of IR-HepG2 cell model was tested. After the IR-HepG2 model was optimally established, the glucose consumption, glycogen content and cell viability were detected after 24 h administration with different concentrations of berberine, baicalin, puerarin and liquiritin by anthrone method, glycerol phosphate oxidase assay and CCK-8 assay respectively. Results The optimal oleic acid-induced concentration was 1 mmol/L and the optimal induced time was 24 h for the IR-HepG2 cell model that could keep stable more than 36 h. Comparing with IR model group, berberine, puerarin and baicalin significantly increased the glucose consumption, whereas liquiritin did not show significant change in the glucose consumption except for 1 μmol/L. Only 160 μmol/L puerarin and 1 μmol/L baicalin significantly inhibited IR-HepG2 cell viability. Moreover, berberine, puerarin, and baicalin significantly elevated the glycogen content; Liquiritin did not change glycogen content significantly. Conclusion The IR-HepG2 cell model could be stably established with 24 h treatment of 1 mmol/L oleic acid. Berberine, puerarin, and baicalin significantly increased the glucose consumption and glycogen content in the IR-HepG2 cells. The results suggest that berberine, baicalin and puerarin maybe perform different pathways of anti-hyperglycemic effects due to different incentives of IR.

國(guó)家自然科學(xué)基金（81460621）；江西省自然科學(xué)基金重點(diǎn)項(xiàng)目（20143ACB20010）