Effect of berberine on endoplasmic reticulum stress PERK apoptosis pathway in HK-2 cells by high fructose
Author(s): ZHANG Yong, HUANG Chun-lai, FU Ting-ting, CHEN Xiao-qing, WANG Li-ping, ZHUANG Yong-ze, Department of Nephrolog y, Fuzhou General Hospital of Nanjing Command
Pages: 6-11
Year: 2017 Issue: 1
Journal: Medical Journal of Chinese People's Liberation Army
Keyword: endoplasmic reticulum stress; kidney tubules; berberine; apoptosis;
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 24 h. The expression of glucose-regulated protein 78(GRP78), CHOP protein and the phosphorylation levels of PERK, e IF2α were tested by Western blotting. The cell cycles were detected by flow cytometry and the apoptosis of cells were detected by TUNEL staining. Results Western 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-e IF2α in group F were significantly higher than those in group F. Compared with group F, GRP78, CHOP, p-PERK and p-e IF2α in group B and T were significantly lower(P<0.05). The expression of GRP78, CHOP, p-PERK and p-e IF2α 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 e IF2α phosphorylation, down-regulated the expression of GRP78, CHOP protein, thus by regulating PERK Pathways to alleviate cell cycle arrest and reduce cell apoptosis.
Pages: 6-11
Year: 2017 Issue: 1
Journal: Medical Journal of Chinese People's Liberation Army
Keyword: endoplasmic reticulum stress; kidney tubules; berberine; apoptosis;
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 24 h. The expression of glucose-regulated protein 78(GRP78), CHOP protein and the phosphorylation levels of PERK, e IF2α were tested by Western blotting. The cell cycles were detected by flow cytometry and the apoptosis of cells were detected by TUNEL staining. Results Western 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-e IF2α in group F were significantly higher than those in group F. Compared with group F, GRP78, CHOP, p-PERK and p-e IF2α in group B and T were significantly lower(P<0.05). The expression of GRP78, CHOP, p-PERK and p-e IF2α 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 e IF2α phosphorylation, down-regulated the expression of GRP78, CHOP protein, thus by regulating PERK Pathways to alleviate cell cycle arrest and reduce cell apoptosis.
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