CAJ | 학술논문

目的：研究p55 TNFR选择性淋巴毒素对心肌细胞氧化应激损伤的保护作用及其机制。方法取Wistar乳鼠制备原代心肌细胞，建立H2 O2致心肌细胞的氧化应激损伤模型，将细胞分为正常组、模型组、rhLTα处理组及rhLTα－Q107 E处理组，流式细胞术检测细胞凋亡，免疫印迹法检测细胞内NF－κB的活化；实时定量RT－PCR法检测抗凋亡分子的转录水平（ Bcl－2、Bcl－X、XIAP）；免疫印迹法检测MnSOD水平；WST法检测CuZn／Mn－SOD活性。结果在体外大鼠心肌细胞的氧化应激损伤模型中，细胞凋亡率明显增高，伴随着Caspase－3活性明显增高；抗凋亡分子Bcl－2、Bcl－xL和XIAP的表达水平均减少；进一步检测MnSOD的蛋白表达水平下降，CuZn／Mn－SOD活性减弱。当加入rhLTα－Q107 E处理后，可逆转上述改变情况，伴随NF－κB的激活。而野生型LTα作用不明显（P＜0．05）。结论 p55TNFR选择性淋巴毒素rhLTα－Q107 E可通过激活NF－κB，上调抗凋亡分子表达和诱导线粒体MnSOD活化，发挥对抗氧化应激保护心肌细胞的作用，且优于野生型的LTα。
목적：연구p55 TNFR선택성림파독소대심기세포양화응격손상적보호작용급기궤제。방법취Wistar유서제비원대심기세포，건립H2 O2치심기세포적양화응격손상모형，장세포분위정상조、모형조、rhLTα처리조급rhLTα－Q107 E처리조，류식세포술검측세포조망，면역인적법검측세포내NF－κB적활화；실시정량RT－PCR법검측항조망분자적전록수평（ Bcl－2、Bcl－X、XIAP）；면역인적법검측MnSOD수평；WST법검측CuZn／Mn－SOD활성。결과재체외대서심기세포적양화응격손상모형중，세포조망솔명현증고，반수착Caspase－3활성명현증고；항조망분자Bcl－2、Bcl－xL화XIAP적표체수평균감소；진일보검측MnSOD적단백표체수평하강，CuZn／Mn－SOD활성감약。당가입rhLTα－Q107 E처리후，가역전상술개변정황，반수NF－κB적격활。이야생형LTα작용불명현（P＜0．05）。결론 p55TNFR선택성림파독소rhLTα－Q107 E가통과격활NF－κB，상조항조망분자표체화유도선립체MnSOD활화，발휘대항양화응격보호심기세포적작용，차우우야생형적LTα。
Objective To investigate the effect of a novel lymphotoxin with selectively binding to p55 tumor necrosis factor receptor (p55TNFR) on myocardial oxidative stress injury , and explore the mechanism.Methods Primary cardiomyocytes were prepared from hearts of neonatalWistar rats , and myocardial oxidative stress injury model was established by hydrogen peroxide ( H2 O2 ) .Then cells were divided into normal group , H2 O2 treatment group, and treatment groups with rhLTαor rhLTα-Q107E. Cell apoptosis was detected by flow cytometry , and the activation of NF -κB was detected by Western blot analysis .Real -time quantitative RT -PCR assay was applied to detect the expression levels of anti-apoptotic molecules (Bcl-2, Bcl-X and XIAP).The expression and activities of MnSOD or CuZn-SOD were detected by Western blot or WST .Results Increased cell apoptosis and the activity of Caspase-3 was observed in the oxidative stress injury model of myocardial cells .The expression of anti-apoptotic protein Bcl -2, Bcl-xL and XIAP were decreased .The expression level and activity of MnSOD were decreased .However , the treatment of rhLTα-Q107 E, but not wild type LTα, reversed these changes , along with the activation of NF -κB.Conclusion This work demonstrates that rhLTα-Q107 E plays a role on the protection against myocardial oxidative stress injury , through the activation of NF-κB, up -regulating the expression of the anti -apoptotic molecules , activating mitochondrial MnSOD, which was better than the wild type LTα.