南方医科大学学报

南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (01): 103-.

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IR-3T3-L1脂肪胰岛素抵抗细胞的建立

聂绪强，杨建文，史海霞，张玉金，张建永，卞卡   

  • 出版日期:2015-01-20 发布日期:2015-01-20

Establishment of a cell model of insulin-resistant 3T3-L1 adipocytes

  • Online:2015-01-20 Published:2015-01-20

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摘要： 目的探讨3T3-L1脂肪胰岛素抵抗细胞（insulin resistant 3T3-L1 adipocytes cell, IR-3T3-L1）最佳的建立条件。方法采用
地塞米松（Dexamethason，DEX）、1-甲基-3-异丁基黄嘌呤（3-isobutyl-methylxanthine, IBMX）和不同浓度的胰岛素（10-8、10-7、
10-6 mol·L-1）诱导3T3-L1前脂肪细胞分化为成熟的脂肪细胞，Oil red O染色确定最佳胰岛素诱导浓度，继而用1 μmol·L-1 DEX
诱导建立IR-3T3-L1 脂肪胰岛素抵抗细胞。采用葡萄糖氧化酶-过氧化物酶（Glucose oxidase-peroxidase, GOD-POD）法对
IR-3T3-L1细胞的最佳诱导时间（24~120 h）及其稳定时间（24~48 h）进行了考察。结果Oil red O染色发现3T3-L1前脂肪细胞
用DEX、IBMX和10-6 mol·L-1胰岛素诱导9 d，>90% 的前脂肪细胞分化为成熟的脂肪细胞；用1 μmol·L-1 DEX培养96 h后，脂肪
细胞葡萄糖消耗率达到最大（P=0.0003）；稳定时间考察发现，在36 h内葡萄糖消耗与对照组比较有统计学意义（P<0.001）。结
论3T3-L1前脂肪细胞在1 μmol·L-1 DEX、0.5 mmol·L-1 IBMX 和10-6 mol·L-1最佳胰岛素浓度作用下诱导分化为成熟的脂肪细
胞后，用1 μmol·L-1 DEX培养96 h即可成功诱导成IR-3T3-L1-IR脂肪胰岛素抵抗细胞，且在36 h内稳定。

Abstract: Objective To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes. Methods
Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10-8, 10-7, and 10-6 mol ·L-1)
were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulinresistant
3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1 μmol·L-1 DEX, and the changes of glucose concentration
in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay. Results Treatment of 3T3-L1 cells
with DEX, IBMX and 10-6 mol·L-1 insulin for 9 days resulted in the differentiation of >90% of the cells into mature adipocytes.
IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 μmol ·L-1 DEX showed significantly increased glucose
consumption (P=0.0003) as compared with the control group at 36 h (P<0.001). Conclusion 3T3-L1 cells can be induced into
mature adipocytes by exposure to 1 μmol·L-1 DEX, 0.5 mmol·L-1 IBMX and 10-6 mol·L-1 insulin. A 96 h exposure to 1 μmol·L-1
DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.