CAJ | 학술논문

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1μmol· 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1μmol· 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1μmol· 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1μmol· 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。
목적：탐토3T3-L1지방이도소저항세포（insulin resistant 3T3-L1 adipocytes cell, IR-3T3-L1）최가적건립조건。방법채용지새미송（Dexamethason，DEX）、1-갑기-3-이정기황표령（3-isobutyl-methylxanthine, IBMX）화불동농도적이도소（10-8、10-7、10-6 mol· L-1）유도3T3-L1전지방세포분화위성숙적지방세포，Oil red O염색학정최가이도소유도농도，계이용1μmol· L-1 DEX유도건립IR-3T3-L1지방이도소저항세포。채용포도당양화매-과양화물매（Glucose oxidase-peroxidase, GOD-POD）법대IR-3T3-L1세포적최가유도시간（24~120 h）급기은정시간（24~48 h）진행료고찰。결과 Oil red O염색발현3T3-L1전지방세포용DEX、IBMX화10-6 mol· L-1이도소유도9 d，>90%적전지방세포분화위성숙적지방세포；용1μmol· L-1 DEX배양96 h후，지방세포포도당소모솔체도최대（P=0.0003）；은정시간고찰발현，재36 h내포도당소모여대조조비교유통계학의의（P<0.001）。결론3T3-L1전지방세포재1μmol· L-1 DEX、0.5 mmol· L-1 IBMX화10-6 mol· L-1최가이도소농도작용하유도분화위성숙적지방세포후，용1μmol· L-1 DEX배양96 h즉가성공유도성IR-3T3-L1-IR지방이도소저항세포，차재36 h내은정。
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 insulin-resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1μmol· L-1 DEX, and the changes of glucose concen-tration 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 1μmol· L-1 DEX, 0.5 mmol· L-1 IBMX and 10-6 mol· L-1 insulin. A 96 h exposure to 1μmol· L-1 DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.