Ginsenoside Rh₃ Induces Barrier-Associated Injury in an In Vitro Blood-Brain Barrier Model with Involvement of MMP2/9

Area of Science:

• Neuroscience
• Pharmacology
• Cell Biology

Background:

• The blood-brain barrier (BBB) is crucial for central nervous system homeostasis.
• Disruption of the BBB is linked to neurological disorders.
• Ginsenoside Rh3 (GRh3), an anticancer compound, has shown neural cell cytotoxicity, raising neurovascular safety concerns.

Purpose of the Study:

• To investigate the effects of GRh3 on an in vitro BBB model.
• To determine the mechanisms underlying GRh3-induced BBB dysfunction.
• To assess the neurovascular safety of GRh3.

Main Methods:

• Utilized a co-culture transwell BBB model with bEnd.3 endothelial cells, pericytes, and C8-D1A astrocytic cells.
• Assessed transendothelial electrical resistance (TEER), lactate dehydrogenase (LDH) release, and cell viability.
• Analyzed tight junction (TJ) protein expression (ZO-1, Claudin-5, Occludin) and matrix metalloproteinase (MMP) levels.
• Employed molecular docking and siRNA-mediated knockdown of MMP2/9.

Main Results:

• GRh3 exposure reduced TEER, increased LDH release, and decreased cell viability.
• GRh3 downregulated TJ proteins (ZO-1, Claudin-5, Occludin) and altered MMP/TIMP1 balance.
• Molecular docking suggested GRh3 interaction with MMP2/9.
• MMP2/9 knockdown partially mitigated GRh3-induced BBB damage.

Conclusions:

• GRh3 impairs BBB integrity, partly via MMP2/9 activation.
• Concerns exist regarding GRh3's neurovascular safety due to cytotoxicity.
• Further in vivo studies are needed to clarify direct barrier effects versus secondary injury.