HO-1高表达对脂肪干细胞在低氧无血清条件下的保护作用及机制  被引量:2

Up-regulation of HO-1 protects ADSCs under serum-free and hypoxic conditions

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作  者:周中新[1] 程轶[2] 代传忠 武维恒[3] 

机构地区:[1]徐州医学院附属医院胸心外科,江苏徐州221002 [2]徐州医学院心血管研究所,江苏徐州221002 [3]徐州医学院第二附属医院心内科,江苏徐州221006

出  处:《中国病理生理杂志》2014年第11期2054-2058,共5页Chinese Journal of Pathophysiology

基  金:江苏省徐州市科技局计划项目(No.XF10C046)

摘  要:目的:探讨诱导内源性血红素氧合酶-1(HO-1)表达对脂肪干细胞(ADSCs)在低氧无血清条件下的保护作用及其可能的分子机制。方法:分离培养SD大鼠脂肪干细胞进行低氧无血清处理,DAPI染色检测细胞凋亡率;Western blotting法分析HO-1、NLRP3、凋亡相关斑点样蛋白(ASC)和cleaved caspase-1的蛋白表达;ELISA法测定培养上清液中白细胞介素-1β(IL-1β)的水平;采用DCFH-DA荧光探针检测细胞内活性氧(ROS)的水平。结果:钴原卟啉诱导ADSCs HO-1蛋白呈剂量依赖性表达,以20μmol/L最为显著;诱导内源性HO-1的表达明显抑制ADSCs在低氧无血清条件下的细胞凋亡率和NLRP3、ASC、cleaved caspase-1的蛋白表达,并减少细胞内ROS和上清液IL-1β的水平;而这种保护效应可被同时给予的HO-1抑制剂锌原卟啉所逆转。结论:诱导内源性HO-1的表达可能通过抑制NLRP3炎症小体的激活以及减少IL-1β的分泌,对低氧无血清条件下ADSCs发挥保护作用。AIM: To investigate the protective effects of endogenous heme oxygenase 1( HO-1) induced by cobalt protoporphyrin( Copp,a HO-1 inducer) on adipose tissue-derived stromal cells( ADSCs) under the condition of serum-free and hypoxia. METHODS: The ADSCs were isolated from SD rat and cultured. The cell apoptotic rate was detected by DAPI staining. The protein expression of HO-1,NLRP3,apoptosis-associated speck-like protein containing a CARD( ASC) and cleaved caspase-1 in ADSCs was messured by Western blotting. IL-1β level in supernatant was determined by ELISA. The level of intracellular reactive oxygen species( ROS) was detected using DCFH-DA. RESULTS: The up-regulation of HO-1 was induced by Co PP in a dose dependent manner and was most significant at 20 μmol / L. The increased expression of HO-1 induced by Co PP significantly reduced the apoptotic rate of ADSCs,intracellular ROS level and IL-1βsecretion,and inhibited the overexpression of NLRP3,ASC and cleaved caspase-1 under serum and oxygen deprivation.These protective effects were reversed by zinc protoporphyrin( Zn PP, an HO-1 inhibitor) given simultaneously.CONCLUSION: The up-regulation of HO-1 expression induced by Co PP plays protective effect on ADSCs under the condition of serum and oxygen deprivation via inhibiting the activation of NLRP3 inflammasome and reducing IL-1β secretion.

关 键 词:脂肪干细胞 血红素氧合酶-1 钴原卟啉 细胞凋亡 

分 类 号:R654.2[医药卫生—外科学]

 

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