High-Throughput Screening Assay Identifies Berberine and Mubritinib as Neuroprotection Drugs for Spinal Cord Injury via Blood-Spinal Cord Barrier Protection

by Selleckchem | Mar 18 2023

Because the breakdown of the blood-brain spinal cord barrier (BBSCB) worsens many central nervous system (CNS) diseases, prevention of BBSCB breakdown has been a major therapeutic target, especially for spinal cord injury (SCI). However, effective drugs that protect BBSCB function have yet to be developed. The purpose of the current study was 1) to develop a high-throughput screening assay (HTSA) to identify candidate drugs to protect BBSCB function, 2) to identify candidate drugs from existing drugs with newly developed HTSA, and 3) to examine the therapeutic effects of candidate drugs on SCI. Our HTSA included a culture of immortalized human brain endothelial cells primed with candidate drugs, stress with H2O2, and evaluation of their viability. A combination of the resazurin-based assay with 0.45 mM H2O2 qualified as a reliable HTSA. Screening of 1,570 existing drugs identified 90 drugs as hit drugs. Through a combination of reproducibility tests, exclusion of drugs inappropriate for clinical translation, and dose dependency tests, berberine, mubritinib, and pioglitazone were identified as a candidate. An in vitro BBSCB functional test revealed that berberine and mubritinib, but not pioglitazone, protected BBSCB from oxygen-glucose deprivation and reoxygenation stress. Additionally, these two drugs minimized BBSCB breakdown 1 day after cervical SCI in mice. Furthermore, berberine and mubritinib reduced neuronal loss and improved gait performance 8 weeks after SCI. Collectively, the current study established a useful HTSA to identify potential neuroprotective drugs by maintaining BBSCB function and demonstrated the neuroprotective effect of berberine and mubritinib after SCI.

Comments：

The purpose of the study was to develop a high-throughput screening assay (HTSA) to identify candidate drugs that can protect the function of the blood-brain spinal cord barrier (BBSCB) and examine the therapeutic effects of these drugs on spinal cord injury (SCI). The researchers developed an HTSA that involved culturing immortalized human brain endothelial cells with candidate drugs, stressing them with H2O2, and evaluating their viability using a resazurin-based assay with 0.45 mM H2O2. They then used this HTSA to screen 1,570 existing drugs and identified 90 hit drugs.

The researchers performed reproducibility tests, exclusion of drugs inappropriate for clinical translation, and dose dependency tests to identify berberine, mubritinib, and pioglitazone as candidate drugs. They then conducted an in vitro BBSCB functional test and found that berberine and mubritinib, but not pioglitazone, protected BBSCB from oxygen-glucose deprivation and reoxygenation stress. They also found that berberine and mubritinib minimized BBSCB breakdown 1 day after cervical SCI in mice and reduced neuronal loss and improved gait performance 8 weeks after SCI.

Overall, the study established a useful HTSA for identifying potential neuroprotective drugs that maintain BBSCB function and demonstrated the neuroprotective effect of berberine and mubritinib after SCI. This research could lead to the development of new treatments for SCI and other CNS diseases that involve BBSCB breakdown.