Efficacy of Bunao Fuyuan decoction on cerebral ischemia and reperfusion injury in vitro.

OBJECTIVE To investigate the protective efficacy of Bunao Fuyuan decoction (BNFY) on cerebral Ischemia/reperfusion (I/R) injury. METHODS The mouse PC12 cells were chosen, and the oxidative-glucose deprivation/re-oxygenation (OGD/R) injury model were established to simulate cerebral I/R injury. Atorvastatin was selected as a positive drug, and a gradient dose of BNFY was given for 6, 12 and 24 h. 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay were used to detect cell viability at each time point. Cell apoptosis was measured by terminal deoxynucleotidyl transferase-mediated dUTP-botin nick end labeling (TUNEL) staining. enzyme linked immunosorbent assay was used to detect the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β and platelet activating factor (PAF). Western blot assay were performed to detect the expression of key regulators [toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB), p-p38 mitogen-activated protein kinase (MAPK) and p-Akt (also known as protein kinase B, PKB)] of cell survival and inflammatory response. RESULTS The results of MTT assay and TUNEL staining assay revealed that BNFY significantly increased cell viability and inhibited cell apoptosis of PC12 cells following OGD/R, respectively. Furthermore, the expression of TNF-α, 1L-6, 1L-1 and PAF were decreased after BNFY treatment. And the results of Western blot assay showed that BNFY downregulated TLR4, NF-κB, p-p38 MAPK expression and upregulated p-Akt expression. CONCLUSION Our findings suggest that BNFY may play a role in protecting OGD/R injured PC12 cells through inhibiting the inflammatory response and cell apoptosis.