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[目的]研究白细胞介素-37(interleukin-37,IL-37)在人肝肿瘤细胞系(HepG2)细胞中的表达,及在炎症因子刺激下,HepG2细胞中IL-37表达水平的改变。[方法]在37℃、5%CO2条件下培养人HepG2细胞,以脂多糖(lipopolysaccharide,LPS)刺激细胞,分别于刺激24h、48h、72h后提取细胞,分别提取细胞总RNA、总蛋白,利用逆转录酶聚合链反应技术(RT-PCR)检测细胞中IL-37-mRNA的表达,利用Western blot技术检测细胞中IL-37蛋白的表达水平。并且在LPS刺激细胞72h后,运用免疫荧光和激光共聚焦技术检测IL-37在细胞中的表达定位。[结果]RT-PCR结果证明HepG2中存在IL-37-mRNA的表达,且随着LPS刺激时间的延长IL-37-mRNA表达增多,0h(无刺激)、24h、48h、72h灰度值分别为0.10±0.008、0.14±0.014、0.22±0.010、0.28±0.014;Western blot结果显示细胞中存在IL-37蛋白的表达,且其表达趋势与IL-37mRNA表达趋势相同,即随着刺激时间延长,HepG2中IL-37蛋白表达增多,由24h、48h、72h分别为0.54±0.16、0.87±0.04、1.51±0.18;免疫荧光共聚焦结果显示IL-37在细胞核和细胞质中均有表达,以细胞核周围表达最为丰富。[结论]人HepG2细胞中存在IL-37表达,炎症因子刺激后,IL-37mRNA及IL-37蛋白表达水平均呈升高,趋势相同;IL-37在HepG2细胞质内和细胞核内均表达,以细胞核周围表达最为丰富。
[Objective] To investigate the expression of interleukin-37 (IL-37) in human hepatoma cell line (HepG2) and the change of IL-37 expression in HepG2 cells stimulated by inflammatory cytokines. [Method] Human HepG2 cells were cultured under the condition of 37 ℃ and 5% CO2, cells were stimulated by lipopolysaccharide (LPS), and the cells were extracted after 24h, 48h and 72h respectively. Total RNA and total protein were extracted, The expression of IL-37-mRNA in cells was detected by reverse transcriptase polymerase chain reaction (RT-PCR), and the expression of IL-37 protein was detected by Western blot. After LPS stimulated 72h, the expression of IL-37 in cells was detected by immunofluorescence and confocal laser scanning microscope. [Result] The results of RT-PCR showed that the expression of IL-37-mRNA existed in HepG2, and the expression of IL-37-mRNA increased with LPS stimulation time, 0h (no stimulation), 24h, 48h, Which was 0.10 ± 0.008,0.14 ± 0.014,0.22 ± 0.010,0.28 ± 0.014. The results of Western blot showed that the expression of IL-37 protein was present in the cells, and its expression tendency was the same as that of IL-37mRNA expression, that is, as the stimulation time prolonged, The expression of IL-37 protein in HepG2 cells increased from 0.54 ± 0.16,0.87 ± 0.04,1.51 ± 0.18 at 24h, 48h and 72h, respectively. Immunofluorescence confocal results showed that IL-37 was expressed in both nucleus and cytoplasm, The most abundant expression. [Conclusion] The expression of IL-37 is present in human HepG2 cells. The expressions of IL-37 mRNA and IL-37 protein are all increased with the same trend after stimulated by inflammatory cytokines. IL-37 is expressed both in cytoplasm and nucleus in HepG2 The most abundant expression around the nucleus.