目的 探討黑樹(shù)莓花色苷（ROAs）對(duì)HepG2細(xì)胞胰島素抵抗（IR）的影響。方法 MTT法檢測(cè)3.75、7.50、15.00、30.00、60.00 μg·mL^-1的ROAs作用24、48 h對(duì)HepG2細(xì)胞存活率的影響，確定ROAs的給藥條件；含高糖（4.5 g·L^-1）、高胰島素（1.0×10^-7、1.0×10^-6、1.0×10^-5、1.0×10^-4 mol·L^-1）的DMEM培養(yǎng)基培養(yǎng)HepG2細(xì)胞24或48 h，采用葡萄糖氧化試劑盒檢測(cè)每孔上清液中葡萄糖水平，計(jì)算每孔細(xì)胞葡萄糖消耗量，確定IR細(xì)胞模型的建立條件；HepG2細(xì)胞分為對(duì)照組、模型組、二甲雙胍（陽(yáng)性藥，0.01 mol·L^-1）組和ROAs（7.5、15.0、25.0、30.0 μg·mL^-1）組，除對(duì)照組外，各組細(xì)胞培養(yǎng)24 h后建立IR模型，模型建立后藥物處理24 h，試劑盒法檢測(cè)葡萄糖消耗量，HE染色評(píng)價(jià)細(xì)胞形態(tài)學(xué)改變。結(jié)果 選擇7.5、15.0、25.0、30.0 μg·mL^-1作為ROAs后續(xù)給藥質(zhì)量濃度，給藥時(shí)間為24 h；HepG2細(xì)胞IR模型建立條件為：含高糖（4.5 g·L^-1）、高胰島素（1.0×10^-4 mol·L^-1）的DMEM培養(yǎng)基培養(yǎng)24 h。與對(duì)照組比較，模型組細(xì)胞葡萄糖的消耗量明顯降低，具有統(tǒng)計(jì)學(xué)差異（P＜0.001）；與模型組比較，質(zhì)量濃度為15.0、25.0、30.0 μg·mL^-1的ROAs組細(xì)胞葡萄糖消耗量均顯著增加，具有統(tǒng)計(jì)學(xué)意義（P＜0.05、0.001）。與對(duì)照組比較，模型組細(xì)胞的形態(tài)不固定，細(xì)胞核圓形，胞漿不成型，脂滴明顯增多；ROAs對(duì)HepG2細(xì)胞的胞漿形態(tài)有改善作用并且使脂滴明顯減少。結(jié)論 ROAs能夠改善高糖、高胰島素誘導(dǎo)的HepG2細(xì)胞IR。

Objective To investigate the effect of Rubus occidentalis anthocyanins (ROAs) on insulin resistance (IR) in HepG2 cells. Methods MTT assay was used to detect the effects of ROAs of 3.75, 7.50, 15.00, 30.00, and 60.00 μg·mL^-1 on the survival rate of HepG2 cells for 24 and 48 h, and to determine the administration conditions of ROAs. HepG2 cells were cultured in DMEM medium containing high glucose (4.5 g·L^-1) and high insulin (1.0×10^-7, 1.0×10^-6, 1.0×10^-5, 1.0×10^-4 mol·L^-1) for 24 or 48 h. The glucose level in the supernatant of each well was detected by glucose oxidation kit, and the glucose consumption of cells in each well was calculated, to establish the conditions of IR cell model. HepG2 cells were divided into control group, model group, metformin (positive drug, 0.01 mol·L^-1) group and ROAs (7.5, 15.0, 25.0, 30.0 μg·mL^-1) group. Except for the control group, the IR model was established after 24 h of cell culture. After the establishment of the model, cells were treated with drugs for 24 h, Glucose consumption was detected by kit method, and morphological changes were evaluated by HE staining. Results 7.5, 15.0, 25.0, 30.0 μg·mL^-1 were selected as the following concentrations of ROAs, and the administration time was 24 h. The IR model of HepG2 cells was established under the following conditions:DMEM medium containing high glucose (4.5 g·L^-1) and high insulin (1.0×10-4 mol·L^-1) for 24 h. Compared with the control group, the consumption of glucose in IR model group was significantly decreased (P < 0.001). Compared with the model group, the glucose consumption of cells in the ROAs groups with concentrations of 15.0, 25.0, and 30.0 μg ·mL^-1 was significantly increased (P < 0.05, 0.001). Compared with control group, the morphology of IR model cells was not fixed, the nucleus was round, the cytoplasm was not formed, and the lipid droplets increased significantly. ROAs could improve the cytoplasmic morphology of HepG2 cells and significantly reduce lipid droplets. Conclusion ROAs can improve the insulin resistance of HepG2 cells induced by high glucose and high insulin.

R282.75

貴州省科技計(jì)劃項(xiàng)目［黔科合基礎(chǔ)-ZK（2021）一般535］；貴州醫(yī)科大學(xué)國(guó)家自然基金培育項(xiàng)目（19NSP074）