夏文燕++许荣++方向南++秦永章++欧阳龙强

[摘要]目的 探讨吡格列酮和高糖对3T3-L1细胞中p-Perilipin 1A蛋白表达的影响。方法 将以3T3-L1前脂肪细胞诱导分化成3T3-L1脂肪细胞，进行分组，设对照组（5 mmol/L葡萄糖）、高糖组（25 mmol/L葡萄糖）、高糖加吡格列酮组（25 mmol/L葡萄糖+100 μmol/L吡格列酮），通过测定培养基中甘油含量作为评价脂肪分解的指标，同时用Western blot检测p-Perilipin 1A蛋白表达。结果 通过甘油试剂盒监测甘油释放量，高糖组较对照组明显增加，差异有统计学意义（P<0.01），而高糖加吡格列酮组的甘油释放量较高糖组明显减少，差异有统计学意义（P<0.01）；同时Western blot检测结果显示，与对照组比较，高糖组p-Perilipin 1A蛋白明显上调，差异有统计学意义（P<0.01），高糖组与高糖加吡格列酮组比较，显著抑制了p-Perilipin 1A蛋白的上调，差异有统计学意义（P<0.01）。结论 吡格列酮可能通过阻止p-Perilipin 1A上调来抑制高糖刺激下的脂肪分解，减少游离脂肪酸，从而改善胰岛素抵抗。

[关键词]吡格列酮；高糖；3T3-L1细胞；p-Perilipin 1A

[中图分类号] R589.2 [文献标识码] A [文章编号] 1674-4721（2017）04（c）-0009-04

[Abstract]Objective To investigate the effects of pioglitazone and high glucose on the expression of p-Perilipin 1A protein in 3T3-L1 cells.Methods 3T3-L1 preadipocytes were induced and differentiated into 3T3-L1 adipocytes and then were classified into three groups：control group （5 mmol/L glucose），high glucose group （25 mmol/L glucose）and high glucose plus Pioglitazone group （25 mmol/L glucose + 100 μmol/L Pioglitazone）.The glycerol content of culture medium was determined and taken as the evaluation index of lipolysis；at the same time，Western blot was used to detect the expression of p-Perilipin 1A protein.Results According to the release amount of glycerol monitored by using the glycerol kit，a more obvious increase was seen in high glucose group than in control group，with statistically significant difference （P<0.01）；while，a more obvious decrease of the release amount of glycerol occurred in high glucose plus Pioglitazone group than that in high glucose group，with statistically significant difference （P<0.01）；at the same time，the p-Perilipin 1A proteinof high glucose group detected by Western blot was significantly increased in comparison with control group，with statistically significant difference（P<0.01）；and the expression of p-Perilipin 1A protein was significantly inhibited in high glucose group in contrast with high glucose plus pioglitazone group，with statistically significant difference （P<0.01）.Conclusion Pioglitazone may prevent the up-regulation of p-Perilipin 1A to inhibit glucose-stimulated lipolysis and decrease free fatty acids，thereby improving the insulin resistance.

[Key words]Pioglitazone；High glucose；3T3-L1 cells；p-Perilipin 1A

目前代谢综合征的发病率正以惊人的速度上升[1]，脂代谢紊乱是代谢综合征的病理特点，因为在血循环中增加了源自脂肪組织的非酯化游离脂肪酸（free fatty acid，FFA）[2]。这些FFA可以引发肌肉、肝脏处胰岛素抵抗[3]。FFA的浓度主要由脂肪细胞的脂肪分解控制。周脂素（PLIN1，Perilipin 1A）能双重调控脂类代谢[4]，Perilipin 1A表达下降和磷酸化上调都能促进脂解，诱发胰岛素抵抗[5]。目前，研究已明确噻唑烷二酮类诱导调节糖、脂代谢的相关蛋白表达而减轻胰岛素抵抗[6]，但是如何调节脂代谢的相关蛋白表达以改善胰岛素抵抗作用的研究尚不多。而本文旨在研究吡格列酮在3T3-L1细胞是否可通过抑制高糖刺激下的脂滴包被蛋白p-Perilipin1A蛋白的表达上调来减少脂肪分解进一步减少FFA，以助于减轻胰岛素抵抗。

1材料与方法

1.1主要试剂

3T3-L1前脂肪细胞株（美国模式培养物集存库提供）、葡萄糖（北京益利精细化学品有限公司）、地塞米松（D4902）、1-甲基-3-异丁基-黃嘌（I5879）、无酚红DMEM培养基……