论文部分内容阅读
目的探讨4-苯基丁酸(4-PBA)对棕榈酸诱导的大鼠RIN-m5F细胞内质网应激(ERS)的效应。方法大鼠RIN-m5F细胞用基础培养基孵育24 h,转为无糖1640培养基,孵育24 h,加入1 mmol/L棕榈酸及不同浓度4-PBA共孵育24 h。QT-PCR(实时荧光定量聚合酶链反应)检测4-PBA处理前后ERS相关因子GRP78、CHOP及caspase3mRNA的表达变化。蛋白免疫印迹方法(Western blotting)测定相应蛋白的表达水平。结果棕榈酸刺激后细胞内GRP78、CHOP及caspase3 mRNA表达水平增加。10 mmol/L 4-PBA处理后,CHOP、caspase3表达量降低;蛋白免疫印迹结果显示,棕榈酸刺激后CHOP、caspase3蛋白表达增加,10 mmol/L4-PBA处理后表达降低。结论 4-PBA可能通过CHOP途径减轻RIN-m5F细胞的ERS,对细胞凋亡具有一定的抑制作用。
Objective To investigate the effect of 4-phenylbutyrate (4-PBA) on palmitate-induced endoplasmic reticulum stress (ERS) in rat RIN-m5F cells. Methods Rat RIN-m5F cells were incubated with basal medium for 24 h and transformed into sugar-free 1640 medium for 24 h. The cells were incubated with 1 mmol / L palmitic acid and different concentrations of 4-PBA for 24 h. The changes of ERS-related factors GRP78, CHOP and caspase3mRNA were detected by QT-PCR (real-time fluorescence quantitative polymerase chain reaction) before and after 4-PBA treatment. Western blotting was used to determine the expression level of the corresponding protein. Results Palmitic acid increased the intracellular GRP78, CHOP and caspase3 mRNA expression levels. After treated with 10 mmol / L 4-PBA, the expression of CHOP and caspase3 decreased. Western blot analysis showed that the expression of CHOP and caspase 3 increased after palmitate stimulation, and decreased after treatment with 10 mmol / L 4-PBA. Conclusions 4-PBA may reduce ERS in RIN-m5F cells through CHOP pathway, which may inhibit apoptosis.