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

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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)：O2 5%，CO2 5%，N2 90%；B 组(过度低氧条件组)：O2 2%，CO2 5%，N2 93%；C 组(低氧并经CoCl2 处理组)：O2 5%，CO2 5%，N2 90%；D 组(低氧条件并经CAY10585 处理组)：O2 5%，CO2 5%，N2 90%。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。

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	杨立群
	屠冠军

关键词 ：髓核细胞, 低氧诱导因子, 凋亡

Abstract：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-1α was induced in different degrees in different conditions.Group A (hypoxia condition group): O2 5%,CO2 5%,N2 90%;group B (over hypoxia condition group): O2 2%,CO2 5%,N2 93%;group C(hypoxia and CoCl2 treatment group): O2 5%,CO2 5%,N2 90%;group D (hypoxia and CAY10585 treatment group): O2 5%,CO2 5%,N2 90%.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-1α protein expression was observed in all groups.The expression of HIF-1α protein in group B and group C was higher than that in group A,the expression of HIF-1α protein 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-1α expression can induce the apoptosis of nucleus pulposus cells.The expression of apoptosis related factors p53,Bax,Bcl-2 and caspase-3 is detected by HIF-1α expression in different degrees,which indicates the correlation between the expression of HIF-1α and the expression of apoptosis related proteins,thus reveals the effect of HIF-1α on the expression of apoptosis related factors in the mitochondrial apoptosis pathway of SD rat nucleus pulposus cell.

Key words： Nucleus pulposus cells Hypoxia inducible factor Apoptosis

基金资助:沈阳医学院科技基金项目（20195067）

引用本文:

李旭；叶放；刘一秀；赵宇；谢鑫；杨立群；屠冠军. 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[J]. 中国当代医药, 2021, 28(1): 10-15.
LI Xu；YE Fang；LIU Yi-xiu；ZHAO Yu； XIE Xin；YANG Li-qun；TU Guan-jun. Effect of hypoxia inducible factor on the expression of related proteins in apoptotic pathway of nucleus pulposus cells in SD rats. 中国当代医药, 2021, 28(1): 10-15.

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