Neuroprotective effects of methyl 3,4-dihydroxybenzoate against H₂O₂-induced apoptosis in RGC-5 cells.
In the present study, we investigated the protective effect of methyl 3,4-dihydroxybenzoate (MDHB) against H2O2-induced apoptosis in RGC-5 cells. The RGC-5 cells were cultured in plates for 24 h, which were then pretreated with dimethyl sulfoxide, different concentrations of MDHB, or probucol for 12 h prior to addition of 300 μM H2O2 for 24 h. The cell viability was detected by MTT assay. The rate of apoptosis, level of lipid peroxidation, and mitochondrial membrane potential (MMP) were detected by flow cytometry. Western blot analysis was also used to measure the expression level of Bcl-2, Bax, caspase 9, and caspase 3 proteins in H2O2-treated RGC-5 cells. Our study showed that the cell viability of RGC-5 cells significantly decreased after treatment with 300 μM H2O2 for 24 h, but MDHB (8, 16, 32 μM) increased RGC-5 cell survival, suppressed the rate of apoptosis, scavenged reactive oxygen species, and restored MMP. MDHB also obstructed H2O2-induced apoptosis by regulating the expression of Bcl-2 and Bax, as well as suppressing the activation of caspase 9 and caspase 3. Our results showed that MDHB is an effective neuroprotective compound that mitigates oxidative stress and inhibits apoptosis in RGC-5 cells.
Department of Pharmacology, School of Medicine, Jinan University, China.
Xing
X
Zhou
Chao-Fen
CF
Su
Zheng
Z
Zhang
Chen-Yu
CY
Wang
Jin-Qi
JQ
Luo
Xiao-Wen
XW
Zhou
Liang
L
Cai
Li
L
Yan
Wei
W
Zhang
Huan-Min
HM
Luo
D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000009:adverse effects*; D062385:Hydroxybenzoates / Q000494:pharmacology* / Q000627:therapeutic use; D015227:Lipid Peroxidation / Q000187:drug effects; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018696:Neuroprotective Agents / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012162:Retinal Degeneration / Q000188:drug therapy; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000473:pathology*; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
C520290:3,4-dihydroxybenzoate; D062385:Hydroxybenzoates; D018696:Neuroprotective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D006861:Hydrogen Peroxide; D053148:Caspase 3; D053453:Caspase 9