Related Experiment Video
Updated: Nov 2, 2025
Oxygen-Glucose Deprivation and Reoxygenation as an In Vitro Ischemia-Reperfusion Injury Model for Studying Blood-Brain Barrier Dysfunction
Published on: May 7, 2015
Protective effect of transient receptor potential melastatin 2 inhibitor A10 on oxygen glucose
Zhuoqun Huang1, Xiafei Yu1, Xingyu Liu1
1Department of Human Anatomy，Histology and Embryology，School of Basic Medicine，Ningxia Medical University，Yinchuan 750004，China.
Abstract:
:To investigate the effect of transient receptor potential melastatin 2 （TRPM2） inhibitor A10 on oxygen glucose deprivation/reperfusion （OGD/R） injury in SH-SY5Y cells．:Human neuroblastoma SH-SY5Y cells were subject to OGD/R injury，and then were divided into blank control group，model control group and A10 group randomly. The cell survival rate was detected by cell counting kit 8 （CCK-8）; the level of cellular reactive oxygen species （ROS） was detected by reactive oxygen detection kit; the mitochondrial membrane potential was detected by tetramethylrhodamine （TMRM） method; the number of apoptotic cells was detected by TUNEL apoptosis assay kit; the protein expression level of cleaved caspase 3 was detected by Western blot．:Compared with 3，20，30，50， has lower cytotoxicity and better inhibition effect on channel activity. Compared with the model control group，ROS level was reduced，the mitochondrial membrane potential was improved，the number of apoptosis cells was reduced ，and the expression of cleaved caspase 3 was significantly reduced in the A10 group（all <0.05）. : A10 can alleviate cell damage after OGD/R by inhibiting TRPM2 channel function，reducing extracellular calcium influx，reducing cell ROS levels，stabilizing mitochondrial membrane potential levels，and reducing apoptosis.
Insights
The TRPM2 inhibitor A10 protects SH-SY5Y cells from oxygen-glucose deprivation/reperfusion injury. A10 reduces reactive oxygen species, stabilizes mitochondrial potential, and decreases apoptosis, demonstrating its neuroprotective effects.
Area of Science:
- Neuroscience
- Cell Biology
- Pharmacology
Background:
- Oxygen-glucose deprivation/reperfusion (OGD/R) triggers neuronal injury.
- Transient Receptor Potential Melastatin 2 (TRPM2) channels are implicated in OGD/R-induced cell damage.
- Developing targeted therapies for OGD/R injury is crucial.
Purpose of the Study:
- To evaluate the efficacy of the TRPM2 inhibitor A10 against OGD/R injury in human neuroblastoma SH-SY5Y cells.
- To elucidate the protective mechanisms of A10 in this cellular model.
Main Methods:
- SH-SY5Y cells were subjected to OGD/R.
- Cell viability was assessed using CCK-8.
- Reactive oxygen species (ROS) levels were measured.
- Mitochondrial membrane potential (MMP) was evaluated using TMRM.
- Apoptosis was quantified via TUNEL assay.
- Cleaved caspase-3 expression was determined by Western blot.
Main Results:
- A10 demonstrated low cytotoxicity and effectively inhibited TRPM2 channel activity.
- Compared to the OGD/R model group, A10 treatment significantly reduced ROS levels.
- A10 treatment improved MMP, decreased apoptotic cell numbers, and lowered cleaved caspase-3 expression.
Conclusions:
- A10 alleviates OGD/R-induced cell damage in SH-SY5Y cells.
- Inhibition of TRPM2 channel function by A10 reduces extracellular calcium influx, ROS production, and apoptosis.
- A10 shows potential as a therapeutic agent for neuroprotection against ischemic injury.
