摘要
目的利用体外细胞模型研究尿毒症毒素对甲酚对单核细胞释放炎症因子的影响及其可能机制。方法采用CCK-8法检测THP-1单核细胞株经20、40、80、160mg/L对甲酚分别处理6、12、24h后细胞增殖能力的变化;根据细胞增殖实验结果,选取40mg/L和80mg/L两个剂量组作为低剂量组和高剂量组,提取细胞RNA和蛋白质,用RT-PCR法、Western blotting检测对甲酚处理24h后THP-1单核细胞株炎症因子TNF-α、抗炎因子IL-10以及炎症相关Toll样受体4(TLR-4)m RNA和蛋白表达的变化。结果对甲酚可抑制THP-1细胞增殖,其抑制作用呈浓度和时间依赖性(P<0.05);RT-PCR及Western blotting检测显示,对甲酚处理的THP-1细胞炎症因子TNF-α释放增加,IL-10释放减少,TLR-4表达增加(P<0.05)。结论对甲酚可抑制THP-1细胞增殖,抑制IL-10抑炎因子表达,并可能通过上调TLR-4的表达,促进促炎因子TNF-α的释放,从而导致微炎症状态。
Objective To evaluate the effect of uremic toxin p-cresol on the release of inflammatory cytokines from monocytes and explore its potential mechanism. Methods CCK-8 was employed to evaluate the proliferation status of monocytes cell-line THP-1 after being exposed to 20, 40, 80, 160mg/L of p-cresol for 6, 12 or 24h, while 40mg/L and 80mg/L were assigned as low-dose group and high-dose group. KNA and protein were extracted. RT-PCR and Western blotting were employed to evaluate the mRNA and protein expression of proinflammatory cytokine TNF-a anti-inflammatory cytokine IL-10, as well as Toll-like receptor-4 （TLK-4） of THP-1 cell exposed to p-cresol for 24h. Results P-cresol depressed the proliferation ability of THP-1 in a dose- and time-dependent manner （P〈0.05）. RT-PCR and Western blotting showed that both TNF-a and TLK-4 were over-expressed in THP-1 cell exposed to p-cresol, while the expression of IL-10 was reduced （P〈0.05）. Conclusion P-cresol may inhibit both the proliferation of TI--IP- 1 cell and the release of IL-10. By up-regulating the expression of TLR-4, p-cresol may stimulate the release of TNF- o~, resulting in microinflammation.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2015年第1期35-39,共5页
Medical Journal of Chinese People's Liberation Army
