CD36在棕榈酸诱导的HepG2细胞炎症反应中的作用

发布时间：2018-10-12 09:38

【摘要】：目的:探讨白细胞分化抗原36(cluster of differentiation 36,CD36)是否参与了棕榈酸诱导的HepG2细胞炎症反应。方法:给予HepG2细胞不同浓度的棕榈酸(0.00、0.08、0.16、0.32、0.64 mmol/L)处理24 h,使用实时荧光定量PCR方法检测CD36和炎症因子的mRNA表达水平,Western blot检测CD36蛋白表达水平。然后利用小RNA干扰技术,构建低表达CD36的HepG2细胞(CD36RNAi HepG2)和对照细胞(NCi HepG2)模型。通过实时荧光定量PCR检测棕榈酸负荷的NCi HepG2和CD36RNAi HepG2中炎症因子的表达情况,荧光探针DCFH DA法检测细胞内活性氧含量,Western blot检测细胞核内的核转录因子-κB的蛋白表达水平。结果:棕榈酸能够上调HepG2细胞中CD36及炎症因子——肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-6(interleukin-6,IL-6)的表达。棕榈酸浓度分别为0.00、0.08、0.16、0.32及0.64 mmol/L时,CD36的mRNA表达相对值分别为(1.001±0.078)、(1.510±0.341)(q=4.763,P=0.007)、(1.780±0.070)(q=7.301,P=0.000)、(2.510±0.141)(q=14.16,P=0.000)及(2.103±0.150)(q=10.34,P=0.000)。TNF-α的mRNA表达相对值分别为(1.000±0.098)、(1.571±0.177)(q=1.598,P=0.141)、(1.725±0.274)(q=2.016,P=0.071)、(2.113±0.341)(q=3.102,P=0.011)及(5.282±0.855)(q=11.940,P=0.000)。IL-6的mRNA表达相对值分别为(0.999±0.047)、(1.791±0.596)(q=1.486,P=0.168)、(2.119±0.294)(q=2.107,P=0.061)、(2.808±0.147)(q=3.398,P=0.007)及(6.916±1.284)(q=11.130,P=0.000)。抑制HepG2细胞中CD36表达,可以显著降低棕榈酸所诱导的细胞内活性氧(reactive oxygen species,ROS)增加和核转录因子-κB在核内分布的增强,降低HepG2细胞内炎症因子表达水平。NCi HepG2、NCi HepG2+棕榈酸组及CD36 RNAi HepG2+棕榈酸组的ROS相对含量分别为(1.000±0.113)、(1.968±0.293)(与NCi组相比,t=6.888,P=0.000)、(0.519±0.129)(与NCi+棕榈酸组相比,t=10.106,P=0.000)。NCi HepG2、NCi HepG2+棕榈酸组及CD36 RNAi HepG2+棕榈酸组的核转录因子-κB的蛋白相对值分别为(0.997±0.093)、(2.443±0.085)(与NC相比,t=19.892,P=0.000)、(1.607±0.107)(与NCi+棕榈酸组相比,t=10.607,P=0.000)。抑制HepG2细胞中CD36表达后,棕榈酸浓度分别为0.08、0.16、0.32及0.64 mmol/L时,NCi HepG2及CD36 RNAi HepG2组的TNF-α的表达水平分别为(1.004±0.113)和(0.185±0.071)(t=10.716,P=0.000);(1.009±0.160)和(0.221±0.028)(t=8.389,P=0.001);(1.008±0.163)和(0.173±0.011)(t=8.780,P=0.001);(1.009±0.163)和(0.147±0.013)(t=9.013,P=0.000)。抑制HepG2细胞中CD36表达后,棕榈酸浓度分别为0.08、0.16、0.32及0.64 mmol/L时,NCi HepG2及CD36 RNAi HepG2组的IL-6的表达水平分别为(1.044±0.347)和(0.207±0.033)(t=4.121,P=0.015);(1.006±0.139)和(0.198±0.007)(t=10.110,P=0.001);(1.001±0.052)和(0.125±0.006)(t=28.443,P=0.000);(1.012±0.188)和(0.114±0.015)(t=8.367,P=0.001)。结论:棕榈酸能够上调HepG2细胞CD36表达,促进HepG2炎症反应;抑制CD36可能通过减少氧化应激、抑制核转录因子-κB信号通路,进而改善棕榈酸诱导的HepG2细胞炎症。
[Abstract]:Aim: to investigate the role of leukocyte differentiation antigen 36 (cluster of differentiation 36 (CD36) in palmitic acid-induced inflammation of HepG2 cells. Methods: HepG2 cells were treated with different concentrations of palmitic acid (0.000. 08 ~ 0.16 ~ 0. 32 ~ 0. 64 mmol/L) for 24 h. The expression level of CD36 and mRNA of inflammatory factors was detected by real-time fluorescence quantitative PCR. CD36 protein expression was detected by, Western blot. Then HepG2 cells (CD36RNAi HepG2) and control cells (NCi HepG2) with low expression of CD36 were constructed by using small RNA interference technique. The expression of inflammatory factors in palmitic acid-loaded NCi HepG2 and CD36RNAi HepG2 was detected by real-time fluorescence quantitative PCR, and the expression of nuclear transcription factor- 魏 B was detected by fluorescence probe DCFH DA method. The active oxygen species (Ros) content in the cells was detected by, Western blot. Results: palmitic acid could up-regulate the expression of CD36, tumor necrosis factor- 伪 (TNF- 伪) and interleukin-6 (interleukin-6,IL-6) in HepG2 cells. 妫曟�堥吀娴撳害鍒嗗埆涓�0.00,0.08,0.16,0.32鍙，

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