微小RNA-375对腰椎间盘突出大鼠JAK2/STAT3信号通路的影响

Effect of microRNA-375 on JAK2/STAT3 signaling pathway in lumbar disc herniation

目的分析微小RNA-375(miR-375)对腰椎间盘突出大鼠JAK激酶2(JAK2)/信号传导及转录激活因子3(STAT3)信号通路的影响。方法选取20只8周龄雄性SD大鼠开展研究,通过硬膜外自体髓核移植法成功建立腰椎间盘突出大鼠模型18例,采用随机数字表法分为A组、B组、C组,每组各6例。取各组大鼠髓核组织,A组和B组采用Lipofectamine 2000脂质体分别转染miR-375模拟物和miR-375阴性对照,C组不转染。3组标本均培养48 h,分别于3、6、12、24、48 h时检测并比较各组细胞凋亡率、迁移率、侵袭个数,并比较培养48 h后各组白细胞介素(IL)-1β、IL-1、IL-6、肿瘤坏死因子-α(TNF-α)水平和JAK2、磷酸化JAK2(p-JAK2)、STAT3、磷酸化STAT3(p-STAT3)水平。结果随着培养时间的延长,各组细胞凋亡率、细胞迁移率均呈现增长趋势,培养3~48 h内A组的细胞凋亡率、细胞迁移率均低于B组和C组;随着培养时间的延长,各组细胞侵袭个数均呈现增多趋势,培养3~48 h内A组的细胞侵袭个数少于B组和C组,差异均有统计学意义(P<0.05)。培养48 h后A组和B组的IL-1β、IL-1、IL-6、TNF-α水平均高于C组,但A组低于B组,差异均有统计学意义(P<0.05)。培养48 h后A组的JAK2、p-JAK2、STAT3、p-STAT3水平均低于B组和C组,且A组低于B组,差异均有统计学意义(P<0.05)。结论miR-375高表达可通过抑制JAK2/STAT3信号通路,抑制间盘细胞侵袭、迁移和凋亡,缓解炎症反应。

Objective To investigate the effect of microRNA-375(miR-375)on JAK kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in lumbar disc herniation.Methods Twenty 8-week-old male SD rats were selected for the study,and 18 cases of lumbar disc herniation rat models were successfully established through epidural autologous nucleus pulposus transplantation.They were divided into groups A,B,and C by random number table,6 cases in each group.Nucleus pulposum tissues of rats in each group were collected.MiR-375 mimics and miR-375 negative control were transfected with Lipofectamine 2000 liposome respectively in groups A and B,while no transfection was performed in group C.The three groups of specimens were cultured for 48 hours.The apoptosis rate,migration rate,and number of invasions in each group were detected and compared at 3,6,12,24,and 48 hours.The levels of interleukin(IL)-1β,IL-1,IL-6,tumor necrosis factor-α(TNF-α)and JAK2,phosphorylated JAK2(P-JAK2),STAT3,phosphorylation STAT3(p-STAT3)were compared after 48 hours of culture.Results With the extension of culture time,the apoptosis rate and cell migration rate of each group showed an increasing trend,and the apoptosis rate and cell migration rate of group A were lower than those in group B and group C within 3-48 h;with the extension of culture time,the number of cell invasion in each group showed an increasing trend,and the number of cell invasion in group A was less than those in group B and group C within 3-48 h,the differences were statistically significant(P<0.05).After 48 hours of culture,the levels of IL-1β,IL-1,IL-6,and TNF-αin group A and group B were higher than those in group C,but group A were lower than those in the group B,the differences were statistically significant(P<0.05).The levels of JAK2,p-JAK2,STAT3 and p-STAT3 in group A were lower than those in group B and group C,and group A were lower than those in the group B,the differences were statistically significant(P<0.05).Conclusion High expression of miR-375 can inhibit the invasion,migration and apoptosis of disc cells and alleviate the inflammatory response by inhibiting JAK2/STAT3 signaling pathway.