Cadmium promotes senescence of annulus fibrosus cells via activation of PI3K/Akt signaling pathway

doi:10.12307/2023.990

Abstract

Abstract: BACKGROUND: Cadmium is a common environmental pollutant, which can damage multiple organs and tissues, such as the kidney and bone, but its effect on annulus fibrosus cells in the intervertebral disc has been less reported.
OBJECTIVE: To investigate the effect of cadmium chloride on the senescence of annulus fibrosus cells and the role of PI3K/Akt signaling pathway.
METHODS: Annulus fibrosus cells from Sprague-Dawley rat intervertebral discs were harvested and passage 3 cells were intervened with different concentrations of cadmium chloride (0, 1, 5, 10, 20 μmol/L). Cell viability and proliferation were detected by cell counting kit-8 assay. Transcriptome sequencing and Kyoto Encyclopedia of Genes and Genomes functional enrichment analysis were performed on annulus fibrosus cells with or without cadmium chloride addition. Passage 3 annulus fibrosus cells were divided into control group, cadmium chloride group and LY294002 group. Cell proliferation rate was detected by EdU method, positive cell rate was detected by senescence-associated β-galactosidase staining, and expressions of senescence-associated proteins (p16, p21 and p53) and p-Akt at protein and mRNA levels were measured by western blot, RT-PCR and immunofluorescence.
RESULTS AND CONCLUSION: 5 μmol/L cadmium chloride could inhibit the proliferation of annulus fibrosus cells. Results from the Kyoto Encyclopedia of Genes and Genomes functional enrichment analysis showed that the main signal transduction pathways included PI3K/Akt, cell cycle and p53 signaling pathways, which were related to cell senescence and proliferation. PI3K/Akt signaling pathways with significant differential expression were selected for validation. Compared with the control group, the EdU-positive rate was significantly decreased in the cadmium chloride group (P < 0.05), while the β-galactosidase-positive rate, the expression of senescence-associated proteins (p16, p21 and p53) and p-Akt significantly increased (P < 0.05). Compared with the cadmium chloride group, the EdU-positive rate and p-Akt expression were significantly decreased in the LY294002 group (P < 0.05), while the β-galactosidase-positive rate and the expression of senescence-associated proteins (p16, p21 and p53) significantly increased (P < 0.05). To conclude, cadmium chloride can regulate the senescence of annulus fibrosus cells by activating the PI3K/Akt signaling pathway, thereby inducing the occurrence and progression of intervertebral disc degeneration.

Key words: cadmium, annulus fibrosus cell, intervertebral disc degeneration, senescence, PI3K/Akt, signaling pathway

CLC Number:

Liu Xin, Hu Man, Zhao Wenjie, Zhang Yu, Meng Bo, Yang Sheng, Peng Qing, Zhang Liang, Wang Jingcheng. Cadmium promotes senescence of annulus fibrosus cells via activation of PI3K/Akt signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(8): 1217-1222.

Figures/Tables 7

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