振动对体外血管内皮细胞中炎症因子及IncRNA MEG3表达的影响

Effects of vibration on expressions of vascular endothelial inflammatory factors and IncRNA MEG3 in vitro

[背景]长时间暴露于振动可导致血管内皮损伤,炎症反应在血管内皮损伤中起着重要作用。研究表明长链非编码RNA(lncRNA)母系表达基因3(MEG3)参与内皮细胞的炎症损伤。[目的]探究振动对体外血管内皮细胞分泌炎症因子及lncRNA MEG3表达的影响。[方法]将人脐静脉内皮细胞(HUVEC)分为振动组和对照组:振动组分别施加63 Hz、6.76 m·s^(-2),200 Hz、5.08 m·s^(-2),250 Hz、4.56 m·s^(-2)频段的振动,每日接振1~4 h,分别暴露1、2 d;对照组除不接触振动外,其他均与实验组相同。采用CCK-8检测不同振动频率和时间对HUVEC活力的影响。采用酶联免疫吸附试验检测细胞和上清液中肿瘤坏死因子-α(TNF-α)、白介素(IL)-8、IL-4、IL-10的浓度。通过实时荧光定量PCR检测lncRNA MEG3的表达水平。[结果]与对照组相比,63 Hz振动1 d组,1.5、3 h后细胞活力上升,2、2.5 h后下降;2 d组,振动1.5 h后细胞活力上升,2、2.5 h后下降。200 Hz振动1 d组,振动2、4 h后细胞活力上升,2.5、3 h后下降;2 d组,振动1.5 h后细胞活力上升,2.5 h后下降。250 Hz振动1 d组,1.5、2.5 h后细胞活力上升,3 h后下降;2 d组,振动1.5 h后细胞活力上升,3 h后下降。在63、200 Hz振动2.5 h,250 Hz振动3 h条件下,与对照组相比:1 d组和2 d组TNF-α、IL-8、IL-4、IL-10在细胞和上清液中浓度均升高;1 d组lncRNA MEG3的表达水平下降,2 d组仅63 Hz振动后lncRNA MEG3的表达水平下降。以上结果均具有统计学意义(P<0.05)。[结论]振动可以诱导体外血管内皮细胞中炎症因子TNF-α、IL-8、IL-4、IL-10水平升高以及lncRNA MEG3表达水平下降。

[Background]Prolonged exposure to vibration can cause vascular endothelial injury,and inflammatory response plays an important role in vascular endothelial injury.Studies have shown that long non-coding RNA(IncRNA)maternally expressed gene 3(MEG3)is involved in regulating the expression of inflammatory injury of endothelial cells.[Objective]To investigate the effects of vibration on the secretion of inflammatory factors and the expression of IncRNA MEG3 by vascular endothelial cells in vitro.[Methods]Human umbilical vein endothelial cells(HUVEC)were divided into two categories:vibration and control.The vibration exposure included 63 Hz(6.76 m·s^(-2)),200 Hz(5.08 m·s^(-2)),and250 Hz(4.56 m·s^(-2))frequency bands,and 1 and 2 d exposure time with 1 to 4 h of daily vibration.The control treatment was the same as the vibration category except that they were not exposed to vibration.CCK-8 was used to detect the effects of different vibration frequencies and time on the viability of HUVEC.The expression levels of tumor necrosis factor-α(TNF-α),inte rleukin-8(IL-8),interleukin-4(IL-4),and interleukin-10(IL-10)in the cells and supernatants were detected by enzyme-linked immunosorbent assay.The expression levels of IncRNA MEG3 were detected by real-time fluorescence quantitative PCR.[Results]Compared with the cells with the control treatment,the cell viability of the 1-day exposure group increased after 1.5 h and 3 h of vibration at 63 H z,while decreased after 2 h and 2.5 h;the cell viability of the 2-day exposure group increased at the frequency of 63 Hz for 1.5 h,but decreased at 2 h and 2.5 h.At the frequency of 200 Hz,the cell viability of the 1-day exposure group increased at 2 h and 4 h,but decreased at 2.5 h and 3 h;the cell viability of the 2-day exposure group increased at 1.5 h and decreased at 2.5 h.For the vibration exposure at frequency of 250 Hz,the cell viability of the 1-day exposure group increased at 1.5 h and 2.5 h,but decreased at 3 h;of the 2-day exposure group,the cell viability increased at 1.5 h and decreased at 3 h.For the exposure settings of 63 and 200 Hz vibration for 2.5 h and 250 Hz vibration for 3 h,and with the control treatment as reference,the expression levels of TNF-α,IL-8,IL-4,and IL-10 in cells and supernatants were increased in the 1 d and 2 d exposures;the expression level of lncRNA MEG3 decreased in the 1 d exposure group;however,for the 2 d exposure,the expression level of lncRNA MEG3 decreased only in the 63 Hz vibration exposure.All of these results were statistically significant(P<0.05).[Conclusion]Vibration could induce an increase in the levels of inflammatory factors TNF-α,IL-8,IL-4,and IL-10 and a decrease in the expression level of lncRNA MEG3 in vascular endothelial cells in vitro.