OBJECTIVE To investigate the effect of Buyang Huanwu Decoction on microglial P2Y12 receptor-mediated RhoA/ROCK inflammatory signaling pathway in mice with cerebral ischemia/reperfusion (I/R) injury.
METHODS The C57BL/6J mice were randomly assigned to the Sham group, MCAO group, BYHW-L (low-dose, 6.5 g·kg-1) group, BYHW-H (high-dose, 26 g·kg-1) group, and BYHW-H+MRS2395 group. The middle cerebral artery occlusion (MCAO) model was constructed, and the neurological impairment of the mice was evaluated by Longa biological score and TTC staining. The phenotypic polarization of microglia towards M1 or M2 was detected by immunofluorescence staining and qPCR. The protein expression levels of factors in RhoA/ROCK pathway mediated by the P2Y12 receptor were detected by Western blot.
RESULTS Compared with MCAO group, Buyang Huanwu Decoction could reduce the neurological impairment (P < 0.05) and infarct volume of I/R injured mice (P < 0.05, P < 0.01), increase the polarization of M1 to M2 in microglia cells (P < 0.05, P < 0.01), inhibit the expression of P2Y12 receptor protein (P < 0.05, P < 0.01), block the P2Y12-mediated RhoA/ROCK pathway and p38 MAPK phosphorylation in the downstream pathway (P < 0.05, P < 0.01), and inhibit the inflammatory damage (P < 0.05, P < 0.01). The P2Y12 receptor inhibitor MRS2395 did not block the anti-inflammatory effect of Buyang Huanwu Decoction.
CONCLUSION Buyang Huanwu Decoction may help to improve brain function after cerebral I/R injury in mice through P2Y12-mediated RhoA/ROCK inflammatory signaling pathway, it may also play an anti-inflammatory role through other pathways, but the other mechanisms need further exploration.
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