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目的观察人肝癌Hep G2细胞脂肪变模型中内质网应激发生的情况,探讨分子伴侣4-苯基丁酸(4-PBA)对该脂肪变模型中内质网应激、氧化应激和凋亡的影响。方法使用0.5、1.0 mmol/L的混合脂肪酸分别干预Hep G2细胞,在不同时间点采用Real-time PCR方法检测内质网应激相关蛋白CHOP和葡萄糖调节蛋白78(GRP78)mRNA的表达水平。以2 mmol/L的4-PBA干预脂肪变性Hep G2细胞,在不同时间点检测CHOP和GRP78 mRNA的表达,氧化应激指标丙二醛(MDA)含量及超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)水平,以及凋亡相关蛋白Caspase-3活性的变化。结果与对照组比较,混合脂肪酸干预组Hep G2细胞内CHOP、GRP78 mRNA的表达水平显著升高(P<0.05)。4-PBA干预可使脂肪变性Hep G2细胞内CHOP和GRP78 mRNA的表达水平下调(P<0.05),MDA含量明显下降,SOD和GSH水平显著升高(P<0.05),Caspase-3活性减低(P<0.05)。结论 Hep G2细胞脂肪变性时发生明显的内质网应激。分子伴侣4-PBA能减轻氧化应激和内质网应激,下调脂肪变性Hep G2细胞Caspase-3活性,减轻细胞损伤。
Objective To observe the occurrence of endoplasmic reticulum stress (ER) in Hep G2 cell steatosis model and to explore the effect of 4-phenylbutyric acid (4-PBA) on endoplasmic reticulum stress, oxidative stress and Effect of apoptosis. Methods Hep G2 cells were treated with 0.5 and 1.0 mmol / L mixed fatty acids respectively, and the expression of endoplasmic reticulum stress-related proteins CHOP and GRP78 mRNA was detected by Real-time PCR at different time points. Hep G2 cells were treated with 2-mmol / L 4-PBA, the expression of CHOP and GRP78 mRNA, the contents of malondialdehyde (MDA) and superoxide dismutase (SOD) and The level of glutathione (GSH), and the activity of apoptosis-related protein Caspase-3. Results Compared with the control group, the levels of CHOP and GRP78 mRNA in Hep G2 cells treated with mixed fatty acid were significantly increased (P <0.05). 4-PBA treatment could down-regulate the expression of CHOP and GRP78 mRNA in steatosis Hep G2 cells (P <0.05), decrease MDA content and increase the activity of SOD and GSH (P <0.05) and decrease the activity of Caspase-3 P <0.05). Conclusion Hep G2 cells undergo significant endoplasmic reticulum stress during steatosis. Molecular chaperone 4-PBA can reduce oxidative stress and endoplasmic reticulum stress, downregulate Caspase-3 activity in steatosis Hep G2 cells, reduce cell damage.