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补体C1q/肿瘤坏死因子相关蛋白3对3T3-L1脂肪细胞脂肪因子表达的影响

Effects of C1q/tumor necrosis factor related protein-3 on the expression of adipokines in 3T3-L1 adipocytes

摘要:

目的:观察脂肪因子补体C1q/肿瘤坏死因子(TNF)相关蛋白3(CTRP3)对3T3-L1脂肪细胞脂联素(APN)、瘦素(LPT)、内脏脂肪素(VFT)及爱帕琳肽(APL)等脂肪因子表达的调节效应,以及胰岛素抵抗对该调节效应的影响。方法以软脂酸诱导胰岛素抵抗的3 T3-L1脂肪细胞模型,分别以10、50、250μg/L CTRP3干预正常3T3-L1脂肪细胞12以及250μg/L CTRP3干预胰岛素抵抗的3T3-L1脂肪细胞12 h。分别通过酶联免疫吸附法( ELISA)及实时定量-聚合酶链反应( RT-PCR)法检测脂肪因子蛋白分泌量及基因表达水平。组间差异采用方差分析,两组间进一步比较采用SNK-q检验。结果250μg/L CTRP3干预正常组APN、LPT、VFT及APL蛋白分泌量较正常对照组分别增加了63.3%、42.9%、57.1%及56.0%( q=8.605、8.526、8.284、8.573,均 P<0.05);10及50μg/L干预组上述脂肪因子蛋白分泌量呈增加趋势,但除50μg/L CTRP3干预组APL蛋白分泌量较对照组显著增加外[(6.2±1.1)比(5.0±0.9)μg/L, q=4.593,P<0.05],其余均差异无统计学意义(均P>0.05);其基因表达变化趋势与此类似,并且在干预浓度为50μg/L时APN、LPT、VFT及APL mRNA表达水平较正常对照组分别增加22.0%、13.0%、20.0%及33.0%( q=6.150、3.987、5.653、9.031,均P<0.05)。与CTRP3(250μg/L)干预正常脂肪细胞相比,CTRP3(250μg/L)干预胰岛素抵抗脂肪细胞APN、LPT、VFT及APL蛋白分泌量分别降低了28.6%、21.0%、24.5%及17.9%( q=6.341、5.969、5.592、4.287,均 P <0.05),基因表达降低了21.6%、17.2%、15.6%及18.9%(q =9.225、7.668、7.066、8.210,均P<0.05)。结论 CTRP3浓度依赖性地增加3T3-L1脂肪细胞APN、LPT、VFT及APL的表达,胰岛素抵抗降低该调节效应。

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abstracts:

Objective To investigate the impacts of C1q/tumor necrosis factor (TNF) related protein-3 ( CTRP3 ) on the expression of adiponectin ( APN ) , leptin ( LPT ) , visfatin ( VFT ) and apelin (APL) in 3T3-L1 adipocytes.The effect of insulin resistance on the impacts was also investigated.Methods The insulin resistant 3T3-L1 adipocytes were induced by palmic acid.There were six groups:normal control group, insulin resistance group, CTRP3(10,50,250 μg/L) treated normal group and CTRP3 (250 μg/L) treated insulin resistance group.The secretion and gene expression of the adipokines were detected by enzyme-linked immunosorbent assay ( ELISA ) and real-time polymerase chain reaction ( RT-PCR ) respectively.One-way ANOVA was used for statistical analysis of the differences among groups and SNK-q test was used for the further comparison between two groups.Results Compared with normal control group , the secretion of APN , LPT, VFT and APL in 250 μg/ml CTRP3 intervention group was increased by 63.3%, 42.9%, 57.1%and 56.0%( q=8.605,8.526,8.284,8.573,all P<0.05) respectively.The secretion of such adipokines had increased trend in 10 and 50 μg/L CTRP3 intervention groups , while only the increase of APL in 50 μg/L CTRP3 intervention group was significant ( ( 6.2 ±1.1 ) vs ( 5.0 ±0.9 )μg/L, q=4.593, P<0.05).The gene expression had the same trend after treatment , and the mRNA relative expression of APN , LPT, VFT and APL at the intervention concentration of 50 μg/L was increased&nbsp;by 22.0%, 13.0%, 20.0%and 33.0%respectively in comparison with that in normal control group ( q=6.150, 3.987, 5.653, 9.031, all P<0.05).Compared with the CTRP3 treated normal group, the protein release of APN , LPT, VFT and APL in CTRP3 treated insulin resistant group were decreased by 28.6%, 21.0%, 24.5%and 17.9%respectively (q=6.341, 5.969, 5.592, 4.287, all P<0.05), and gene expression were decreased by 21.6%, 17.2%, 15.6% and 18.9% respectively ( q =9.225, 7.668, 7.066, 8.210, all P<0.05).Conclusion CTRP3 may dose-dependently increase the secretion and gene expression of APN, LPT, VFT and APL in 3T3-L1 adipocytes.Insulin resistance may inhibit this effect.

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作者: 李新 [1] 杨杪 [1] 吴玉文 [1] 孙苏欣 [1] 孙家忠 [1]
期刊: 《中华糖尿病杂志》2014年7期 538-542页 ISTICPKUCSCD
栏目名称: 论著
DOI: 10.3760/cma.j.issn.1674-5809.2014.07.013
发布时间: 2014-08-28
基金项目:
中央高校基本科研业务专项资金
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