Ginsenosides induce extensive changes in gene expression and inhibit oxidative stress-induced apoptosis in human lens epithelial cells

Abstract

Background: The effect of ginsenosides on the growth and apoptosis of human lens epithelial (HLE) B3 cells exposed to H 2 O 2 was investigated. In addition, the effect of ginsenosides on gene expression in HLE-B3 cells was analyzed using microarray assays to determine its molecular mechanism. Methods: HLE-B3 cells were treated with 1.75 M H 2 O 2 in the presence or absence of 5, 10 or 20 μM ginsenosides. Cell viability and apoptosis were examined by MTT assays and flow cytometry, respectively, at 24 to 120 h after the treatment. Furthermore, HLE-B3 cells were treated with 20 μM ginsenosides for 8 days and total RNA was isolated and analyzed using the Affymetrix GeneChip Array. Principal component analysis was performed to visualize the microarray data. Results: Addition of ginsenosides significantly alleviated the growth inhibitory effect of H 2 O 2 on HLE-B3 cells and the percentage of viable cells was increased by more than 3 folds. Flow cytometric analysis showed that 6.16 ± 0.29% of H 2 O 2 -treated HLE-B3 cells were early apoptotic cells, and the percentage was reduced to 4.78 ± 0.16% (P < 0.05) in the presence of 20 μM ginsenosides. Principal component analysis revealed that ginsenoside caused extensive changes in gene expression in HLE-B3 cells. A total of 6219 genes showed significant differential expression in HLE-B3 cells treated with ginsenoside; among them, 2552 (41.0%) genes were significantly upregulated, whereas 3667 (59.0%) genes were significantly downregulated. FOXN2, APP and RAD23B were the top three upregulated genes while WSB1, PSME4 and DCAF7 were the top three downregulated genes in HLE-B3 cells treated with ginsenosides. Conclusion: Ginsenosides induce extensive changes in the expression of genes involved in multiple signaling pathways, including apoptotic signaling pathway and DNA damage response signaling pathway. Ginsenosides alleviate H 2 O 2 -induced suppression of the growth of HLB cells and inhibit H 2 O 2 -induced apoptosis of HLB cells.

lncRNA TUG1 regulates Smac/DIABLO expression by competitively inhibiting miR-29b and modulates the apoptosis of lens epithelial cells in age-related cataracts

“…Therefore, hydrogen peroxide stimulation is often used to construct an in vitro oxidative stress model of HLEC to simulate the occurrence of cataracts. [36] Moreover, we used H 2 O 2 to construct an in vitro model of HLEC apoptosis and found that the expression of TUG1 and Smac increased in this model, while miR29b decreased, which was similar to the expression trend in the anterior capsule of ARC, suggesting that our cataract cell model in vitro was successfully constructed. This provides an experimental basis for further study on the function of TUG1 in this model.…”

Proteomics-based evaluation of the mechanism underlying vascular injury via DNA interstrand crosslinks, glutathione perturbation, mitogen-activated protein kinase, and Wnt and ErbB signaling pathways induced by crotonaldehyde

“…The catalytic subunit of the upregulated (FC = 1.86) DNA polymerase epsilon (protein accession number D3DXI9) acts as the major leading-strand DNA polymerase to repair DNA damage [ 23 ], whereas the upregulation (FC = 1.80) of alpha-2-macroglobulin (protein accession number P01023) is related to inhibition of DNA fragmentation, which is a hallmark of CRA-induced apoptosis [ 24 ]. Upregulation (FC = 2.16) of proteasome activator complex subunit 4 (protein accession number F8WBH5) is related to signal transduction by p53 class mediator, resulting in cell cycle arrest to repair DNA damage [ 25 ], whereas downregulation (FC = 0.65) of keratin 23 (protein accession number Q8TC04) implies that CRA exposure weakened the expression of genes involved in DNA damage response, mainly molecules of the double strand break repair and homologous recombination pathways [ 26 ]. These new indicators among the differentially expressed proteins are closely associated with the DNA repair process (Table 1 ).…”