LncRNA SNHG14通过调控miR-372-3p对OGD/R诱导的星形胶质细胞增殖、凋亡和炎性反应的影响

Effects of long non-coding RNA-SNHG14 on the proliferation,apoptosis and inflammation of oxygen glucose deprivation/reoxygenation-induced astrocytes in rats by regulating miR-372-3p

目的 探讨长链非编码RNA(LncRNA) SNHG14通过调控miR-372-3p对氧糖剥夺/复氧(OGD/R)诱导的星形胶质细胞增殖、凋亡和炎性反应的影响。方法 体外分离大鼠星形胶质细胞并建立OGD/R损伤细胞模型,分为Control组、OGD/R组、OGD/R+si-NC组、OGD/R+si-SNHG14组、OGD/R+miR-NC组、OGD/R+miR-372-3p组、OGD/R+si-SNHG14+anti-miR-NC组和OGD/R+si-SNHG14+anti-miR-372-3p组。qRT-PCR检测SNHG14和miR-372-3p的表达量;MTT检测细胞活力;流式细胞术检测细胞凋亡;Western blot检测Bcl-2和Bax蛋白表达;ELISA检测TNF-α、IL-1β的表达;双荧光素酶报告实验验证SNHG14和miR-372-3p的关系。结果 与Control组比较,OGD/R组SNHG14表达量、细胞凋亡率、Bax蛋白表达、TNF-α、IL-10显著升高(P<0.05),miR-372-3p表达量、细胞活力、Bcl-2蛋白表达显著降低(P<0.05)。与OGD/R+si-NC组比较,OGD/R+si-SNHG14组的内细胞凋亡率、Bax蛋白表达、TNF-α、IL-10显著降低(P<0.05);细胞活力、Bcl-2蛋白表达显著升高(P<0.05)。与OGD/R+miR-NC组比较,OGD/R+miR-372-3p组内细胞活力、Bcl-2蛋白表达显著升高(P<0.05),细胞凋亡率、Bax蛋白表达、TNF-α、IL-10显著降低(P<0.05)。与OGD/R+si-SNHG14+anti-miR-NC组比较,OGD/R+si-SNHG14+anti-miR-372-3p组内细胞活力、Bcl-2蛋白表达显著降低(P<0.05),细胞凋亡率、Bax蛋白表达、TNF-α、IL-10显著升高(P<0.05)。结论 敲低SNHG14通过上调miR-372-3p对OGD/R诱导的星形胶质细胞损伤具有一定的保护作用。

Objective To investigate the effects of long non-coding RNA (LncRNA) SNHG14 on the proliferation,apoptosis and inflammation of oxygen glucose deprivation/reoxygenation (OGD/R)-induced astrocytes in rats by regulating miR-372-3p.Methods The astrocytes of rats were isolated in vitro and the OGD/R injury cell models were established,and the cells were divided into control group,OGD/R group, OGD/R+si-NC group,OGD/R+si-SNHG14 group,OGD/R+miR-NC group,OGD/R+miR-372-3p group,OGD/R+si-SNHG14+anti-miR-NC group and OGD/R+si-SNHG14+anti-miR-372-3p group.The qRT-PCR was used to detect the expression levels of SNHG14 and miR-372-3p;MTT detected cell viability;flow cytometry detected cell apoptosis;Western Blot detected the expression levels of Bcl-2 and Bax protein;ELISA detected the expression levels of TNF-α and IL-1β;the dual luciferase report experiment was performed to analyze the correlation between SNHG14 and miR-372-3p.Results Compared with those in control group,the expression levels of SNHG14,the rate of apoptosis,the expression levels of Bax protein,TNF-α,and IL-10 in OGD/R group were significantly increased,however,the expression levels of miR-372-3p,cell viability,and Bcl-2 protein expression levels were significantly decreased (P<0.05).Compared with those in OGD/R+si-NC group,the internal cell apoptosis rate,and expression levels of Bax protein,TNF-α,and IL-10 in OGD/R+si-SNHG14 group were significantly decreased,however,the cell viability and Bcl-2 protein expression levels were increased significantly (P<0.05).Compared with those in OGD/R+miR-NC group,the cell viability and Bcl-2 protein expression levels in OGD/R+miR-372-3p group were significantly increased,but,the apoptosis rate,and expression levels of Bax protein,TNF-α,IL-10 were significantly decre3ased (P<0.05).Compared with those in OGD/R+si-SNHG14+anti-miR-NC group,the cell viability and Bcl-2 protein expression in OGD/R+si-SNHG14+anti-miR-372-3p group were significantly decreased,however,the apoptosis rate,and expression levels of Bax protein,TNF-α,IL-10 were increased significantly (P<0.05).Conclusion The knockdown of SNHG14 has a certain protective effects on astrocyte damage induced by OGD/R through up-regulating the espression of miR-372-3p.