Title:Protective Effects of Astragalus Membranaceus and Ligustrazine on Rat Brain Microvascular Endothelial Cell Injury after Oxygen-Glucose Deprivation/ Reoxygenation by Suppressing the PKCδ/MARCKS Pathway
Volume: 24
Issue: 7
Author(s): Xialin Tang, Huajun Wang, Huihuan Chen, Shuangxi Sun, Hongxia Chen and Ruihuan Pan*
Affiliation:
- The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou,China
Keywords:
Astragalus membranaceus, injury, ligustrazine, oxygen–glucose deprivation/reoxygenation, PKC δ/MARCKS, rat
brain microvascular endothelial cell.
Abstract:
Aim and Objective: Cell death is a main pathological change in brain ischemia. Astragalus
membranaceus (Ast) and ligustrazine (Lig), as traditional Chinese herbs, have a protective effect
against ischemia-reperfusion injury. We aim to find whether the underlying protective mechanism
of Astragalus membranaceus and ligustrazine against Oxygen–glucose deprivation/reoxygenation
(OGD/R) -induced injury in RBMECs is related to PKCδ/MARCKS pathway.
Materials and Methods: OGD/R preconditioning was instituted in rat brain microvascular endothelial
cells (RBMECs). The survival and apoptosis of RBMECs were detected by a Cell Counting
Kit-8 and TUNEL staining; PKCδ/MARCKS and MMP9 expression were examined by immunofluorescence,
western blot and quantitative real-time PCR.
Results: OGD/R stimulation significantly increased RBMEC apoptosis, whereas Ast+Lig, Rottlerin
or Ast+Lig+Rottlerin treatment evidently reduced cellular apoptosis and increased cell viability
(P <0.05). Furthermore, Ast+Lig, Rottlerin or Ast+Lig+Rottlerin treatment significantly reduced
mRNA expression levels of PKCδ/MARCKS and MMP9 (P <0.05), compared to OGD/R control
group. Moreover, Ast+Lig, Rottlerin or Ast+Lig+Rottlerin treatment evidently reduced protein expression
levels of PKCδ, MMP9, and MARCKS (P <0.05), compared to OGD/R control group, detected
by western blotting or immunofluorescence.
Conclusion: The administration of Astragalus membranaceus and ligustrazine protected RBMECs
against OGD/R-induced apoptosis. PKCδ/MARCKS and MMP9 expression were significantly increased
after OGD/R stimulation, while Astragalus membranaceus and ligustrazine treatment evidently
suppressed. Collectively, Astragalus membranaceus and ligustrazine play protective effects
against OGD/R-induced injury in RBMECs through regulating PKCδ/MARCKS pathway to inhibit
MMP9 activation.
