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Ginsenosides induce extensive changes in gene expression and inhibit oxidative stress-induced apoptosis in human lens
Zhewen Wang1, Shiping Zhou2, Xiaoqing Hu3
1Department of Ophthalmology, First Hospital of Jilin University, No.1 Xinmin Avenue, Changchun, 130021, China.
Background:
The effect of ginsenosides on the growth and apoptosis of human lens epithelial (HLE) B3 cells exposed to H2O2 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 H2O2 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 H2O2 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 H2O2-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 H2O2-induced suppression of the growth of HLB cells and inhibit H2O2-induced apoptosis of HLB cells.
Insights
Ginsenosides protect human lens epithelial cells from hydrogen peroxide-induced damage by promoting cell viability and inhibiting apoptosis. This study reveals ginsenosides
Area of Science:
- Ophthalmology
- Cell Biology
- Pharmacology
Background:
- Investigated the impact of ginsenosides on human lens epithelial (HLE) B3 cells exposed to hydrogen peroxide (H₂O₂).
- Analyzed the molecular mechanisms underlying ginsenoside's effects using microarray assays to examine gene expression changes.
Purpose of the Study:
- To determine how ginsenosides affect the growth and apoptosis of HLE-B3 cells under oxidative stress.
- To elucidate the molecular mechanisms of ginsenosides' protective effects through gene expression analysis.
Main Methods:
- HLE-B3 cells were treated with H₂O₂ and varying concentrations of ginsenosides.
- Cell viability and apoptosis were assessed using MTT assays and flow cytometry.
- Gene expression profiling was conducted using Affymetrix GeneChip Arrays and analyzed via principal component analysis.
Main Results:
- Ginsenosides significantly increased cell viability (over 3-fold) and reduced apoptosis in H₂O₂-treated HLE-B3 cells.
- Microarray analysis revealed extensive changes in gene expression, with 6219 genes differentially expressed.
- Key upregulated genes included FOXN2, APP, and RAD23B, while WSB1, PSME4, and DCAF7 were downregulated.
Conclusions:
- Ginsenosides modulate gene expression in pathways related to apoptosis and DNA damage response.
- Ginsenosides effectively counteract H₂O₂-induced growth suppression and apoptosis in HLE-B3 cells.
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