摘要
目的观察游离脂肪酸（FFA）对人成熟脂肪细胞中miR-335表达的调节作用。方法体外培养人前体脂肪细胞，待生长融合后，采用1一甲基-3异丁基黄嘌呤（MIX）、胰岛素、地塞米松、罗格列酮方案诱导人前体脂肪细胞分化为成熟的脂肪细胞，使用含1mmol／L混合FFA的前脂肪细胞培养基（PAM）进行干预，分别培养4、8、24h后收集细胞，同时设未干预的人成熟脂肪细胞为对照组，采用实时荧光定量PCR技术检测成熟脂肪细胞中miR-335的相对表达量。结果对照组细胞中4、8、24hmiR-335的相对表达量与0h相比差异均无统计学意义（P均〉0．05）。使用snRU6作为内参时，FFA干预人成熟脂肪细胞4、8、24h后miR-335的相对表达量分别为9．03-t-O．31、9．85±2．41和11．23±0．62，与对照组0h相比，差异均有统计学意义（P均〈0．05）；使用miR-103作为内参时，同等时间点miR-335的相对表达量分别为4．73±0．60、5．38±1．25和4．57±0．52，与对照组0h相比，差异均有统计学意义（P均〈0．05）。结论FFA可明显上调人成熟脂肪细胞中miR-335的表达，为进一步探索miR-335在脂肪细胞中的功能提供了一定的理论依据。
Objective To explore the role of free fatty acids （FFA） on expression of miR-335 in human ma- tured adipocytes. Methods in order to induce differentiation, confluent human pre-adipocytes （ day 0） were subse- quently cultured in serum-free PAM containing 50 nmol/L insulin, 100 nmol/L dexamethasone,0.5 mmol/L 3-isobutyl- 1-methylxanthine, and 100 μmol/L rosiglitazone. Then human matured adipocytes（ day 16） were treated with 1 mmol/L FFA cocktail composed of lauric acid, myristic acid, linoleic acid, oleic acid, and arachidonic acids for 4,8 and 24 hours. Meanwhile, untreated cells were collected as control group. Total RNA from these adipocytes were extracted and the levels of miR-335 expression were evaluated by real-time PCR. Results The expression of miR-335 at 4,8 and 24 hours showed no statistical significance when compared to 0 hour in untreated matured adipocytes （ all P 〉 0.05 ）. The relative expression of miR-335 after the intervention of FFA in human matured adipocytes were 9.03 ± 0.31,9.85 ± 2.41 and 11.23±0. 62, respectively at 4,8 and 24 hours when used with snRU6 for normalization, and there was statistical signi-ficance compared with 0 hour in control group （ all P 〈0. 05 ）. The levels of miR-335 were 4.73 ± 0.60, 5.38±1.25 and 4.57±0.52 at the same time point when used with miR-103 for normalization, and there was statisti- cal significance compared with 0 hour in control group （ all P 〈 0.05）. Conclusions FFA exert a positive effect on the miR-335 expression in human matured adipocytes ,which provide the basis for the further study about the role of miR- 335 in human adipocytes.
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2013年第19期1464-1466,共3页
Chinese Journal of Applied Clinical Pediatrics
基金
国家自然科学基金（81100618）
江苏省医学创新团队项目（LJ201108）
南京市科技发展计划（201104013）
江苏省普通高校研究生科研创新计划（CXLX12-0565）
