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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

Author(s):

Xialin Tang, Huajun Wang, Huihuan Chen, Shuangxi Sun, Hongxia Chen and Ruihuan Pan*   Pages 1 - 10 ( 10 )

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 blot-ting or immunofluorescence.

Conclusions: 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.

Keywords:

Astragalus membranaceus, Injury, Ligustrazine, Oxygen–glucose deprivation/reoxygenation, PKC δ/MARCKS, Rat brain microvascular endothelial cell.

Affiliation:

The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou, The Second Institute of Clinical Medicine, Guangzhou University of Chinese Medicine, Guangzhou



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