咖啡酸苯乙酯通过Nrf2介导的丝裂原活化蛋白激酶途径上调肝星状细胞系T6中的抗氧化剂水平。

Caffeic acid phenethyl ester up-regulates antioxidant levels in hepatic stellate cell line T6 an Nrf2-mediated mitogen activated protein kinases pathway.

作者信息

Yang Ning, Shi Juan-Juan, Wu Feng-Ping, Li Mei, Zhang Xin, Li Ya-Ping, Zhai Song, Jia Xiao-Li, Dang Shuang-Suo

机构信息

Ning Yang, Juan-Juan Shi, Feng-Ping Wu, Mei Li, Xin Zhang, Ya-Ping Li, Song Zhai, Xiao-Li Jia, Shuang-Suo Dang, Department of Infectious Diseases, the Second Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an 710004, Shannxi Province, China.

出版信息

World J Gastroenterol. 2017 Feb 21;23(7):1203-1214. doi: 10.3748/wjg.v23.i7.1203.

Abstract

AIM

To investigate the antioxidant effect of caffeic acid phenethyl ester (CAPE) in hepatic stellate cell-T6 (HSC-T6) cells cultured and the potential mechanisms.

METHODS

HSC-T6 cells were cultured and treated with various concentrations of CAPE for 24, 48 and 72 h, respectively. Cell proliferation was investigated using the MTT assay, and cell ultrastructural alterations were observed by transmission electron microscopy. Flow cytometry was employed to investigate the effects of CAPE on apoptosis and the levels of reactive oxygen species in HSC-T6 cells cultured . An enzyme immunoassay instrument was used to evaluate antioxidant enzyme expression. The effect on α-smooth muscle actin was shown using immunofluorescence. Gene and protein levels of Nrf2, related factors, and mitogen activated protein kinases (MAPKs), in HSC-T6 cells were detected using RT-PCR and Western blot, respectively.

RESULTS

CAPE inhibited the proliferation and activation of HSC-T6 cells cultured . CAPE increased the antioxidant levels and the translocation of Nrf2 from the cytoplasm to the nucleus in HSC-T6 cells. Moreover, the phosphorylation of MAPKs in cells decreased in response to CAPE. Interestingly, CAPE-induced oxidative stress in the cells was significantly attenuated by pretreatment with MAPKs inhibitors.

CONCLUSION

CAPE inhibits cell proliferation and up-regulates the antioxidant levels in HSC-T6 cells partly through the Nrf2-MAPKs signaling pathway.

摘要

目的

研究咖啡酸苯乙酯(CAPE)对培养的肝星状细胞-T6(HSC-T6)的抗氧化作用及其潜在机制。

方法

培养HSC-T6细胞,分别用不同浓度的CAPE处理24、48和72小时。采用MTT法检测细胞增殖,通过透射电子显微镜观察细胞超微结构变化。运用流式细胞术研究CAPE对HSC-T6细胞凋亡和活性氧水平的影响。使用酶免疫分析仪评估抗氧化酶表达。通过免疫荧光显示对α-平滑肌肌动蛋白的影响。分别采用RT-PCR和蛋白质免疫印迹法检测HSC-T6细胞中Nrf2、相关因子和丝裂原活化蛋白激酶(MAPK)的基因和蛋白水平。

结果

CAPE抑制培养的HSC-T6细胞的增殖和活化。CAPE提高了HSC-T6细胞的抗氧化水平,并使Nrf2从细胞质向细胞核转位。此外,细胞中MAPK的磷酸化因CAPE而降低。有趣的是,MAPK抑制剂预处理可显著减轻CAPE诱导的细胞氧化应激。

结论

CAPE部分通过Nrf2-MAPK信号通路抑制HSC-T6细胞增殖并上调其抗氧化水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/674b/5323445/ac35707071cd/WJG-23-1203-g001.jpg

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