细胞因子信号转导抑制因子3(SOCS3)对视神经损伤大鼠视网膜神经节细胞存活的影响  

Suppressors of cytokine signaling 3 inhibits cell viability of retinal ganglion cells after optic nerve injury

作　　者：党晓洁 任梅[1] 朱江[1] 许治国[1] DANG Xiao-Jie REN Mei ZHU Jiang XU Zhi-Guo

机构地区：[1]西安交通大学医学院附属广仁医院眼科,西安市第四医院眼科,陕西省眼科诊疗中心,陕西省西安市710004

出　　处：《眼科新进展》2017年第3期225-229,共5页Recent Advances in Ophthalmology

摘　　要：目的研究细胞因子信号转导抑制因子3(suppressor of cytokine signaling 3,SOCS3)对视神经损伤大鼠视网膜神经节细胞(retinal ganglion cells,RGCs)存活的影响,并探讨其潜在的分子机制。方法采用视神经横断手术构建大鼠视神经损伤模型,术后分离RGCs。实验分为视神经损伤组(optic nerve transection,ONT组)和假手术组。采用Western blot和RT-PCR检测SOCS3在两组细胞中的表达。随后将SOCS3 siRNA分别转染假手术组和ONT组RGCs,实验进一步分为空白对照组、阴性对照组和SOCS3沉默组。CCK8和MTT法检测细胞存活,Hoechst 33342荧光染色法和流式细胞技术检测细胞凋亡。进一步将mTOR siRNA和SOCS3 siRNA共转染RGCs,检测细胞存活和细胞凋亡。结果视神经损伤3d后,ONT组SOCS3表达水平显著高于假手术组(P=0.049),且随损伤时间延长而升高。与空白对照组相比,SOCS3沉默可显著提高视神经损伤后RGCs的存活率[空白对照组与SOCS3沉默组分别为(49.47±7.35)%和(73.24±8.70)%],降低细胞凋亡率[2组分别为(27.25±0.75)%和(10.96±1.07)%]和细胞生长抑制率[2组分别为(23.06±1.43)%和(10.65±1.77)%]。Hoechst染色也表明SOCS3沉默可改善视神经损伤诱导的细胞凋亡。同时,SOCS3沉默可显著提高视神经损伤后2周mTOR活性标志蛋白pS6的表达;且与单独沉默SOCS3相比,共同沉默mTOR和SOCS3可降低细胞存活率,提高细胞生长抑制率和细胞凋亡率。结论SOCS3沉默可以通过上调损伤后期mTOR的活性而促进损伤RGCs的存活并抑制其凋亡。Objective To explore the effect and potential mechanism of suppressors of cytokine signaling 3 （SOCS3） knockout on cell viability and apoptosis of retinal ganglion cells （RGCs） after optic nerve injury.Methods The optic nerve transection was used to construct optic nerve injury model of rats,and RGCs were isolated after optic nerve injury.The experimental animals were divided into optic nerve transection injury （ONT） group and sham-operation （Sham） group.The expression of SOCS3 in RGCs was detected by Western blot and RT-PCR in each group.Subsequently,SOCS3 siRNA was transfected into RGCs of Sham group and ONT group,and the experimental were further subdivided into blank control group,negative control group and SOCS3 silence groups.Cell viability was measured by CCK8 and MTT methods.Apoptosis was detected by Hoechst 33342 staining and flow cytometry.Furthermore,the mTOR siRNA and SOCS3 siRNA were co-transfected into RGCs,and cell viability and apoptosis were detected.Results The expression of SOCS3 was dramatically increased at 3 days after injury in the ONT group when compared with Sham group （P=0.049）,and it showed an increased tendency gradually along with the extension of injury time.Compared with the blank control in the ONT group,SOCS3 silence markedly promoted cell viability [（49.47±7.35）% vs.（73.24±8.70）%],reduced cell growth inhibition [（27.25±0.75）% vs.（10.96±1.07）%] and apoptosis [（23.06±1.43）% vs.（10.65±1.77）%].The result of Hoechst 33342 staining indicated that SOCS3 silence ameliorated the cell apoptosis induced by ONT.In addition,SOCS3 silence significantly improved pS6 expression at 2 weeks after injury,and mTOR and SOCS3 co-silence reduced cell viability,increased cell growth inhibition and apoptosis compared with SOCS3 silence group after injury.Conclusion SOCS3 silence promotes injury-induced cell viability of RGCs and suppresses injury-induced apoptosis of RGCs via up-regulating mTOR activity in the later period of injury.

关 键 词：细胞因子信号转导抑制因子 MTOR 视神经损伤 视网膜神经节细胞 增殖 

分 类 号：R774[医药卫生—眼科]