低氧诱导因子对SD大鼠髓核细胞凋亡路径中相关蛋白表达的影响  

Effect of hypoxia inducible factor on the expression of related proteins in apoptotic pathway of nucleus pulposus cells in SD rats

作　　者：李旭[1] 叶放[1] 刘一秀[1] 赵宇[1] 谢鑫[2] 杨立群 屠冠军[4] LI Xu;YE Fang;LIU Yi-xiu;ZHAO Yu;XIE Xin;YANG Li-qun;TU Guan-jun(Department of Foot and Ankle Surgery,Shenyang Medical College Affiliated Central Hospital,Liaoning Province,Shenyang 110024,China;Functional Experiment Center,Liaoning University of Traditional Chinese Medicine,Liaoning Province,Shenyang 110847,China;Liaoning Research Institute of Family Planning,China Medical University,Liaoning Province,Shenyang 110031,China;Department of Orthopedics,the First Hospital of China Medical University,Liaoning Province,Shenyang 110001,China)

机构地区：[1]沈阳医学院附属中心医院足踝外科,辽宁沈阳110024 [2]辽宁中医药大学机能实验中心,辽宁沈阳110847 [3]中国医科大学辽宁省计划生育研究所,辽宁沈阳110031 [4]中国医科大学第一临床医院骨科,辽宁沈阳110001

出　　处：《中国当代医药》2021年第1期10-15,共6页China Modern Medicine

基　　金：沈阳医学院科技基金项目(20195067)。

摘　　要：目的探讨低氧诱导因子-1α(HIF-1α)对SD大鼠髓核细胞凋亡路径中相关蛋白表达的影响。方法原代培养SD大鼠髓核细胞,施加不同条件诱导髓核细胞不同程度表达HIF-1α,A组(低氧条件组):O25%,CO25%,N290%;B组(过度低氧条件组):O22%,CO25%,N293%;C组(低氧并经CoCl2处理组):O25%,CO25%,N290%;D组(低氧条件并经CAY10585处理组):O25%,CO25%,N290%。同时检测各组细胞凋亡情况;Western Blot检测HIF-1α、p53、Bax、caspase3、Bcl-2蛋白的表达情况;分析HIF-1α、p53、Bax、Bcl-2及caspase3变化与凋亡率的相关性。结果各组中均能见到HIF-1α蛋白有所表达,B组和C组中HIF-1α蛋白表达高于A组,C组中HIF-1α蛋白表达最高,D组最低,差异有统计学意义(P<0.05);p53、Bax、caspase3蛋白的表达趋势是一致的,在C组中最高,高于A及B组,其中B组蛋白的表达高于A组,而D组中的蛋白表达最低(P<0.05);Bcl-2与Bax表达趋势相反,C组中最低,低于A组,D组最高,高于A组,差异有统计学意义(P<0.05)。流式细胞仪所得的结果显示,C组中细胞的凋亡率最高,而D组最低,组间两两比较,差异有统计学意义(P<0.05)。HIF-1α、p53、Bax、Bcl-2及caspase3变化与凋亡率均呈正相关性(P<0.05)。结论HIF-1α表达程度的增加可以诱发髓核细胞的凋亡程度也相应增加,通过HIF-1α在不同程度表达下检测凋亡相关因子p53、Bax、Bcl-2、caspase3的表达情况,说明了HIF-1α表达与凋亡相关蛋白表达的相关性,从而揭示了HIF-1α对SD大鼠髓核细胞线粒体凋亡路径中相关凋亡因子表达的影响情况。Objective To explore the effect of hypoxia inducible factor-1α(HIF-1α)on the expression of related proteins in apoptotic pathway of nucleus pulposus cells in SD rats.Methods The nucleus pulposus cells of SD rats were primary cultured.The expression of HIF-1αwas induced in different degrees in different conditions.Group A(hypoxia condition group):O25%,CO25%,N290%;group B(over hypoxia condition group):O22%,CO25%,N293%;group C(hypoxia and CoCl2 treatment group):O25%,CO25%,N290%;group D(hypoxia and CAY10585 treatment group):O25%,CO25%,N290%.The apoptosis of each group was detected at the same time.Western Blot was used to detect the expression of HIF-1α,p53,Bax,caspase3 and Bcl-2 protein.The relationship between the changes of HIF-1α,p53,Bax,Bcl-2 and caspase3 and apoptosis rate was analyzed.Results HIF-1αprotein expression was observed in all groups.The expression of HIF-1αprotein in group B and group C was higher than that in group A,the expression of HIF-1αprotein in group C was the highest,and the lowest in group D,the differences were statistically significant(P<0.05).The expression trends of p53,Bax,and caspase3 proteins were the same,the highest in group C and higher than those in group A and B,the protein expression in group B was higher than that in group A,while the protein expression in group D was the lowest(P<0.05).The expression trend of Bcl-2 and Bax was opposite,the lowest in group C,lower than that in group A,the highest in group D,higher than that in group A,the differences were statistically significant(P<0.05).The results obtained by flow cytometry showed that the apoptotic rate of cells in group C was the highest,while that of group D was the lowest,there was a statistically significant difference between the two groups(P<0.05).The changes of HIF-1α,p53,Bax,Bcl-2 and caspase3 were positively correlated with the apoptosis rate(P<0.05).Conclusion The increase of HIF-1αexpression can induce the apoptosis of nucleus pulposus cells.The expression of apoptosis related factors p53,Bax,Bcl-2 an

关 键 词：髓核细胞 低氧诱导因子 凋亡 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]