Astragalus saponins inhibit high glucose-induced autophagy in human retinal pigment epithelial cells by regulating protein kinase R-like endoplasmic reticulum kinase pathway

doi:10.12092/j.issn.1009-2501.2019.04.001

Abstract

Abstract:

AIM: To investigate the effects of astragalus saponins on the regulation of PERK pathway in high glucose-induced autophagy in human retinal pigment epithelial cells. METHODS: Human retinal pigment epithelial cells were cultured in vitro. The experiment was divided into NC group (5.5 mmol/L glucose), HG group (30 mmol/L glucose), and Ast group (30 mmol/L glucose+2.5, 5, 10 μg/mL astragalus saponins). The activity of ARPE-19 cells was detected by MTT. The apoptotic rate was detected by flow cytometry. The expression levels of autophagy-related proteins and key factors of PERK pathway were detected by Western blot. The effect on the level of autophagy was determined by Western blot analysis with the addition of PERK-specific inhibitor GSK2606414. RESULTS:Compared with the normal control group, the activity of ARPE-19 cells in the high glucose group was decreased. The apoptotic rate was increased, the expression level of autophagy-related protein LC3 was significantly increased. The expression level of p62 protein was significantly decreased. The protein levels of GRP78, Perk and CHOP, the key factors of PERK pathways, were significantly up-regulated, and the difference was statistically significant (P＜0.05). Compared with the high glucose group, the activity of ARPE-19 cells in the astragalus saponins group was increased. The apoptotic rate was decreased, the expression level of LC3 protein was down-regulated, the expression level of p62 protein was up-regulated, and the protein levels of GRP78, Perk and CHOP were significantly decreased. The concentration dependence was statistically significant (P＜0.05). PERK-specific inhibitor GSK2606414 was able to further inhibit the expression of LC3 protein in the ARPE-19 cells of the astragalus saponins group and up-regulate the expression of p62 protein, the difference was statistically significant (P＜0.05). CONCLUSION: Astragalus saponins can inhibit the autophagy of human retinal pigment epithelial cells induced by high glucose, and its mechanism is related to the inhibition of protein kinase R-like endoplasmic reticulum kinase pathway.

Key words: astragalus saponins, protein kinase R-like endoplasmic reticulum kinase pathway, human retinal pigment epithelial cells, autophagy, diabetic retinopathy

CLC Number:

YU Jing, ZHAO Hailan, CHENG Kaiyao, SHEN Pingyu, HUANG Jinlin. Astragalus saponins inhibit high glucose-induced autophagy in human retinal pigment epithelial cells by regulating protein kinase R-like endoplasmic reticulum kinase pathway[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(4): 361-368.

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