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目的探讨苦瓜总皂苷对3T3-L1胰岛素抵抗脂肪细胞葡萄糖摄取量、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的影响。方法采用MTT法确定苦瓜总皂苷浓度,利用油红“O”染色法鉴定脂肪细胞的形成,用地塞米松处理脂肪细胞复制胰岛素抵抗细胞模型,添加苦瓜总皂苷作用24 h后,运用实时荧光定量逆转录多聚酶联反应(RT-q PCR)检测脂肪细胞中TNF-α和IL-6的m RNA表达,酶标记免疫吸附测定法(ELISA)检测培养液中TNF-α和IL-6蛋白含量变化。结果选定用于实验的苦瓜总皂苷的作用时间为24 h,浓度为苦瓜高剂量组(5 mg·m L~(-1))、苦瓜低剂量组(2.5 mg·m L~(-1));诱导分化3T3-L1细胞12 d形成成熟细胞;地塞米松处理3T3-L1脂肪细胞48 h胰岛素抵抗脂肪细胞模型复制成功;苦瓜总皂苷干预3T3-L1胰岛素抵抗脂肪细胞后,葡萄糖摄取量显著增高(P<0.01),TNF-α和IL-6的表达和蛋白分泌显著降低(P<0.01),并呈浓度依赖性。结论苦瓜可以改善胰岛素抵抗细胞模型的胰岛素抵抗状态,与降低炎症因子表达相关。
Objective To investigate the effect of total saponin of Momordica charantia on glucose uptake, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in 3T3-L1 insulin resistance adipocytes. Methods The total saponin of Momordica charantia L. was determined by MTT method. The formation of adipocytes was identified by the method of oil red “O” staining. The cells were treated with dexamethasone to replicate the model of insulin resistance. After the total saponin was added for 24 h, The mRNA expression of TNF-α and IL-6 in adipocytes was detected by quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), and the levels of TNF-α and IL-6 in culture medium were detected by enzyme-linked immunosorbent assay Variety. Results The total saponin of Momordica charantia was selected for experiment for 24 hours, with high concentration of bitter gourd (5 mg · m L -1) and low dose of bitter gourd (2.5 mg · m L -1 ); 3T3-L1 cells were induced to differentiate into mature cells 12 days later; 3T3-L1 adipocytes were treated with dexamethasone for 48 h and adipocytes were replicated successfully. After intervention of 3T3-L1 insulin-resistant adipocytes, total glucose uptake (P <0.01). The expressions of TNF-α and IL-6 and protein secretion were significantly decreased (P <0.01) in a concentration-dependent manner. Conclusion Bitter melon can improve the insulin resistance of insulin resistance cell model, and reduce the expression of inflammatory cytokines.