解放军医学杂志2017，Vol.42Issue(1)：6-11,6.DOI:10.11855/j.issn.0577-7402.2017.01.02
小檗碱对果糖诱导人肾小管细胞内质网应激PERK凋亡通路的影响
Effect of berberine on endoplasmic reticulum stress PERK apoptosis pathway in HK-2 cells by high fructose
摘要
Abstract
Objective To investigate the effect of berberine on endoplasmic reticulum stress PERK apoptosis pathway in HK-2 cells by high fructose.Methods HK-2 cells were grown in DMEM/F12, containing 10% fetal bovine serum (FBS) and divided randomly into four groups: normal control group (Group C); Fructose group (Group F): it contains 25mmol/L fructose culture; Berberine group (Group B): 25mmol/L fructose + 10μmol/L berberine treatment group; TUDCA group (Group T):25mmol/L fructose +2μmol/L TUDCA culture group; Cells were collected after culturing 24h. The expression of glucose-regulated protein 78 (GRP78), CHOP protein and the phosphorylation levels of PERK, eIF2α were tested by Western blotting. The cell cycles were detected by flow cytometry and the apoptosis of cells were detected by TUNEL staining.ResultsWestern blotting showed that the expression of GRP78 and CHOP protein in group F was significantly higher than that in group C, and the levels of p-PERK and p-eIF2α in group F were significantly higher than those in group F. Compared with group F, GRP78, CHOP, p-PERK and p-eIF2α in group B and T were significantly lower (P<0.05). The expression of GRP78, CHOP, p-PERK and p-eIF2α in group B had no significant difference. Flow cytometry and TUNEL staining showed that the cell viability of fructose group was significantly lower than that of C group, and the apoptotic index was significantly higher than that of group C (P<0.05). Compared with group F, the activity of HK-2 cells in group B and T significantly increased, otherwise the apoptosis index significantly decreased, and the difference was statistically significant (P<0.05). Compared with group T, the viability index and apoptosis rate of group B had no significant difference (P>0.05).Conclusion Persistent high fructose can activate the intracellular PERK pathway in HK-2 cells, causing endoplasmic reticulum stress. Berberine can inhibit the fructose-induced PERK and eIF2α phosphorylation, down-regulated the expression of GRP78, CHOP protein, thus by regulating PERK Pathways to alleviate cell cycle arrest and reduce cell apoptosis.关键词
内质网应激/肾小管/细胞凋亡/小檗碱Key words
endoplasmic reticulum stress/kidney tubules/berberine/apoptosis分类
医药卫生引用本文复制引用
张勇,黄春来,付婷婷,陈小青,王丽萍,庄永泽..小檗碱对果糖诱导人肾小管细胞内质网应激PERK凋亡通路的影响[J].解放军医学杂志,2017,42(1):6-11,6.基金项目
国家自然科学基金(81373837) （81373837）
福建省自然科学基金(2014J01371)This work was supported by the National Natural Science Foundation of China (81373837) and the Natural Science Foundation of Fujian Province (2014J01371) （2014J01371）
