【摘要】 目的 探究长链非编码RNA (long non-coding RNA, lncRNA) LINC02863协同微管亲和调节激酶2 (microtubule affinity-regulating kinase 2, MARK2)诱发炎症反应在促进慢性肾衰竭(chronic renal failure, CRF)中的作用机制。方法 应用HK-2细胞建立CRF细胞模型，细胞转染后分成阳性对照组(CRF模型细胞)、LINC02863过表达组(CRF模型细胞+LINC02863过表达基因)、LINC02863沉默组(CRF模型细胞+LINC02863沉默基因);阴性对照组为常氧培养的HK-2细胞。逆转录聚合酶链式反应(reverse transcription polymerase chain reaction, RT-PCR)测定LINC02863和MARK2表达；细胞计数试剂盒(cell counting kit 8,CCK-8)法测定各组细胞活力；流式细胞术检测各组细胞凋亡率，酶联免疫吸附分析(enzyme-linked immunosorbent assay, ELISA)测定各组细胞肌酐、尿素氮表达，RT-PCR测定肿瘤坏死因子α(tumor necrosis factor alpha, TNF-α)、白细胞介素6(interleukin 6, IL-6)等炎性因子表达，Western blot法测定MARK2、核因子κB(nuclear factor kappa B,NF-κB)、白细胞介素1β(interleukin 1 beta, IL-1β)和p65蛋白表达。结果 与阴性对照组比较，阳性对照组细胞LINC02863基因表达、MARK2基因表达、细胞凋亡率、肌酐、尿素氮、TNF-α、IL-6水平及MARK2、NF-κB、IL-1β和p65蛋白表达显著升高，细胞活力显著降低(P均<0.05);过表达LINC02863基因后，LINC02863过表达组细胞LINC02863基因表达、MARK2基因表达、细胞凋亡率、肌酐、尿素氮、TNF-α、IL-6水平及MARK2、NF-κB、IL-1β和p65蛋白表达较阳性对照组显著升高，细胞活力较阳性对照组显著降低(P均<0.05);沉默LINC02863基因后，LINC02863沉默组细胞LINC02863基因表达、MARK2基因表达、细胞凋亡率、肌酐、尿素氮、TNF-α、IL-6水平及MARK2、NF-κB、IL-1β和p65蛋白表达较阳性对照组显著降低，细胞活力较阳性对照组显著升高(P均<0.05)。结论 lncRNAs LINC02863和MARK2在CRF中异常高表达，过表达LINC02863基因可协同促进MARK2表达，但沉默LINC02863基因表达可协同拮抗MARK2表达，减轻炎症反应，改善CRF症状，值得临床进一步研究并推广。更多还原

【Abstract】 Objective To investigate the mechanism of long non-coding RNA(lncRNA) LINC02863 cooperates with microtubule affinity-regulating kinase 2(MARK2) inducing inflammatory responses in the progression of chronic renal failure(CRF).Methods A cell model of CRF was established using HK-2 cells. After cell transfection, the cells were divided into the following groups: positive control group(CRF model cells), LINC02863 overexpression group(CRF model cells+LINC02863 overexpression gene) and LINC02863 silencing group(CRF model cells+LINC02863 silencing gene), the negative control group consisted of HK-2 cells cultured under normal oxygen conditions. The expressions of LINC02863 and MARK2 were measured by reverse transcription polymerase chain reaction(RT-PCR); cell viability was assessed using the cell counting kit 8(CCK-8) assay; apoptosis rates were detected by flow cytometry; creatinine and urea nitrogen expressions were measured by enzyme-linked immunosorbent assay(ELISA). The expressions of inflammatory factors such as tumor necrosis factor alpha(TNF-α) and interleukin 6(IL-6) were determined by RT-PCR, the expressions of MARK2, nuclear factor kappa B(NF-κB), interleukin 1 beta(IL-1β) and p65 proteins were assessed by Western blot.Results Compared with the negative control group, the positive control group showed significantly higher expression of LINC02863 and MARK2, increased cell apoptosis rate, elevated levels of creatinine, urea nitrogen, TNF-α, IL-6 as well as higher protein expressions of MARK2, NF-κB, IL-1β and p65, cell viability was significantly reduced(all P<0.05); after overexpressing LINC02863, the LINC02863 overexpression group exhibited significantly higher levels of LINC02863, MARK2 gene expression, cell apoptosis, creatinine, urea nitrogen, TNF-α, IL-6, and protein levels of MARK2, NF-κB, IL-1β and p65 compared to the positive control group, while cell viability significantly decreased(all P<0.05). After silencing LINC02863, the LINC02863 silencing group showed significantly lower levels of LINC02863 and MARK2 gene expression, reduced apoptosis rate, and lower levels of creatinine, urea nitrogen, TNF-α, IL-6 and protein expressions of MARK2, NF-κB, IL-1β and p65, along with a significant increase in cell viability compared to the positive control group(all P<0.05).Conclusion lncRNAs LINC02863 and MARK2 are aberrantly upregulated in CRF. Overexpression of LINC02863 enhances the expression of MARK2 and promotes inflammation, while silencing LINC02863 reduces MARK2 expression and alleviates the inflammatory response, improving CRF symptoms. These findings warrant further clinical investigation and potential application.更多还原