[High Homocysteine Level Promotes Autophagy and Apoptosis of Mouse Hippocampal HT22 Cells Through the Notch1/Hes1 Signaling Pathway]

Overview

Journal Nan Fang Yi Ke Da Xue Xue Bao

Specialty General Medicine

Date 2023 Nov 7

PMID 37933657

Authors

J Zhang

J He

X Mi

X Xu

Y Tian

R Yan

Affiliations

Soon will be listed here.

Abstract

Objective: To explore the mechanism of neuronal injury caused by hyperhomocysteinemia.

Methods: Mouse hippocampal HT22 cells were treated with homocysteine (Hcy, 100 μmol/L), Hcy+folic acid+vitamin B12 (100+fv group) or folic acid+vitamin B12 (0+fv group), and the changes in cell autophagy and apoptosis were detected using transmission electron microscope (TEM) and flow cytometry. The expressions of Hes1, Hes5, Notch1, Jagged1, Bcl-2, Bax, P62 and LC3 in the treated cells were detected with Western blotting and real-time PCR.

Results: Treatment with Hcy for 48 h significantly increased the number of dead cells in HT22 cell cultures. Flow cytometry showed that the percentage of apoptotic cells was significantly higher in cells treated with Hcy alone than in other treatment groups (<0.05). TEM revealed obvious mitochondrial swelling and vacuolation and increased autophagy in Hcy-treated cells. Western blotting showed that the Bax/Bcl-2 ratio was significantly higher in Hcy-treated cells than in the blank control cells and cells in 100+fv group (<0.05). The Hcy-treated cells showed a significantly lower relative expression of P62 than the blank control cells (<0.05), a higher LC3II/LC3I ratio than the cells in the blank control and 100+fv groups (<0.05), and lower expressions of HES1, HES5, Notch1 and Jagged1 proteins than the blank control cells (<0.05). Interference with Hes1 siRNA significantly lowered the expression levels of Hes1 and Jagged1 without obviously affecting Notch1 expression in HT22 cells (>0.05).

Conclusion: High Hcy levels promote autophagy and apoptosis and down-regulate Hes1 and Jagged1 expressions in HT22 cells.

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