CAJ | 학술논문

目的观察促红细胞生成素(EPO)对人视网膜色素上皮(hRPE)细胞抗过氧化氢(H2O2)损伤的影响.方法以传代培养hRPE细胞为研究对象,采用800μmol/L H2O2作用3 h建立hRPE细胞损伤模型,分为正常对照组、单纯损伤组、重组人EPO(rhEPO)治疗组.治疗组又根据加入rhEPO浓度不同分为10、20、40、60 IU/ml亚组.免疫组织化学方法检测细胞中核因子kappa B(NF-kB)的活化情况,比色法测定细胞脂质过氧化产物丙二醛(MDA)含量及乳酸脱氢酶(LDH)细胞释放率.结果 H2O2可使培养液中MDA及LDH释放率上升,单纯损伤组与正常对照组相比,差异有统计学意义(tLDH=29.746,tMDA=20.426,P＜0.05);各治疗组与单纯损伤组比较,MDA及LDH释放率均有显著降低,差异有统计学意义(t10IU=5.770,t 20IU=12.774.t40IU=19.818,t60IU=24.833,P＜0.05;MDA t10IU=5.345,t10IU=10.278,t40IU=18.571,t60IU=20.247,P＜0.05);各治疗组相关分析显示,rhEPO浓度与LDH细胞释放率及MDA含量呈负相关(r=0.976,P=0.024;r=0.968,P=0.032);rhEPO浓度与NF-kB核移位率呈正相关(r=0.998,P=0.002);NF-kB核移位率与MDA含量呈负相关(r=-0.954,P=0.046).结论 EPO能有效地拮抗H2O2对hRPE细胞的脂质过氧化损害,其机制可能与活化NF-KB有关.
목적 관찰촉홍세포생성소(EPO)대인시망막색소상피(hRPE)세포항과양화경(H2O2)손상적영향.방법 이전대배양hRPE세포위연구대상,채용800μmol/L H2O2작용3 h건립hRPE세포손상모형,분위정상대조조、단순손상조、중조인EPO(rhEPO)치료조.치료조우근거가입rhEPO농도불동분위10、20、40、60 IU/ml아조.면역조직화학방법검측세포중핵인자kappa B(NF-kB)적활화정황,비색법측정세포지질과양화산물병이철(MDA)함량급유산탈경매(LDH)세포석방솔.결과 H2O2가사배양액중MDA급LDH석방솔상승,단순손상조여정상대조조상비,차이유통계학의의(tLDH=29.746,tMDA=20.426,P＜0.05);각치료조여단순손상조비교,MDA급LDH석방솔균유현저강저,차이유통계학의의(t10IU=5.770,t 20IU=12.774.t40IU=19.818,t60IU=24.833,P＜0.05;MDA t10IU=5.345,t10IU=10.278,t40IU=18.571,t60IU=20.247,P＜0.05);각치료조상관분석현시,rhEPO농도여LDH세포석방솔급MDA함량정부상관(r=0.976,P=0.024;r=0.968,P=0.032);rhEPO농도여NF-kB핵이위솔정정상관(r=0.998,P=0.002);NF-kB핵이위솔여MDA함량정부상관(r=-0.954,P=0.046).결론 EPO능유효지길항H2O2대hRPE세포적지질과양화손해,기궤제가능여활화NF-KB유관.
Objective To investigate the protective effect and mechanism of erythropoietin(EPO)on injury of human retinal pigment epithelial(hRPE)cell induced by hydrogen peroxide(H2O2).Methods Take subcultured hFRPE cells as study target.They were treated with 800μmol/L of H2O2 for3 hours to establish the eell injury model.The cultured cells were divided into three groups:control group,simply injury group and therapeutic group which again divided into 10 IU/ml,20 IU/ml,40 IU/ml,60IU/ml subgroups according to the concentration of recombinant human erythropoietin(rhEPO).NF-kBwas measured by immunohistochemistry.The content of Malondialdehyde(MDA)which was the productof cellular lipid peroxidation and the releasing rate of lactate dehydrogenase(LDH)were estimated bychromatometry. Results H2O2 could elevate the level of MDA and the releasing rate of LDH,comparedsimply injury group with control group,the differences were significant.(tLDH=29.746 tMDA=20.426,P＜0.05);Compared all of therapeutics groups with simply injury group,the releasing rate of MAD and LDHwere decreased obviously,the differences were significant.(LDH t10IU=5.770,t20IU=12.774,t40IU=19.818,t60IU=24.833,P＜0.05;MDA t10IU=5.345,t20IU=10.278,t40It/=18.571,t60IU=20.247,P＜0.05);negative correlation with the relation rate of LDH and the content of MDA(r=-0.976,P=0.024;r=the content of MDA(r=-0.954,P=0.046). Conclusion EPO can protect hFRPE ceils from the injuryof H2O2,the mechanism may be related to the activation of NF-KB.