Objective To endoplasmic reticulum stress cell model of human umbilical vein endothelial cells(HUVECs), to estoblish observe the effects of coptisine on the growth activity of HUVECs cells, and expression levels of CHOP and GRP78 gene and protein.
Methods HUVECs were induced by tunicamycin(TM) at a concentration of 10 μg/mL for 18 h, and then treated with coptisine at different concentrations for 24 h.The CCK8 was used to detect the effects of different concentrations of coptisine on the activity of HUVECs and activity of HUVECs induced by TM.The mRNA expression levels of CHOP and GRP78 in each group were detected using RT-PCR.The protein expression levels of CHOP and GRP78 in each group were detected using Western blotting and cellular immunofluorescence.
>Results With the concentration of coptisine increasing, the coptisine in the range of 1-20 μmol/L had no effect on the activity of HUVECs.After 24 hours, the concentration of coptisine in the range of 20 μmol/L played a protective effect on the activity of HUVECs induced by TM(P < 0.01).The mRNA expression levels of CHOP and GRP78 in HUVECS increased after 18 hours of TM induction(P < 0.01), after 24 hours of continuous induction with coptisine, it was found that coptisine could inhibit the mRNA expressions of CHOP and GRP78 in TM-induced HUVECs(P < 0.01).The protein expression levels of CHOP and GRP78 in HUVECs increased after 18 hours of TM induction(P < 0.01), and after 24 hours of continuous induction with coptisine, it was found that coptisine could inhibit the protein expression levels of CHOP and GRP78 in TM-induced HUVECs(P < 0.01).The results of cellular immunofluorescence staining were similar to those of Western blotting.
Conclusions Coptisine can alleviate the excessive ER stress induced by TM in HUVECs, and improve the activity of damaged cells.Coptisine can improve the excessive ER stress response of HUVECs by down-regulating the expressions of CHOP and GRP78.
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