Rubescensine Attenuates Early Brain Injury After Subarachnoid Hemorrhage By Inhibiting NLRP3 Inflammasome Formation

Purpose:To investigate the effect of oridonin on the formation of inflammatory bodies in NLRP3 after subarachnoid hemorrhage.Materials and methods:One hundred and twelve male C57BL/6J mice were randomly divided into 6 groups: sham operation group(sham group),model group(SAH group),solvent group(SAH+vehicle group),model + oridonin 5mg/kg group(SAH+oridonin5mg/kg group),model + oridonin 10mg/kg group(SAH+oridonin10mg/kg group)and model +oridonin 20mg/kg group(SAH+oridonin20mg/kg group).The mouse SAH model was established by internal carotid artery puncture.Except sham operation group and model group,different concentrations of oridonin were injected into 30 min after operation,and dimethyl sulfoxide was injected intraperitoneally in solvent group.24 hours later,the balance beam score,modified Garcia score,SAH grade and brain edema were measured.The apoptosis of neurons and the activation of microglia were observed by immunofluorescence.The protein expressions of NLRP3 inflammatory bodies,ASC,caspas-1,IL-1 ?,IL-18 and apoptosis-related proteins Bax and caspase-3 were detected by Westernblot.The experimental results were analyzed by statistical software.Result:1.The results of neurological function score,SAH classification: balance beam score and modified Garcia score showed that the balance beam score and modified Garcia score in the model group and solvent group were significantly lower than those in the sham operation group.Compared with the solvent group,the balance beam score and modified Garcia score in the model + oridonin high and low dose groups were significantly higher than those in the solvent group,and the effect was more obvious in the oridonin 10mg/kg group.Compared with the solvent group,the model + oridonin high and low dose groups could significantly reduce the SAH grade(P < 0.05).2.Brain water content: compared with the sham operation group,the brain water content in the model group and the solvent group increased significantly(P < 0.05).Compared with the solvent group,the high and low dose group of model + oridonin could reduce the brain water content.the effect in the oridonin 10mg/kg group was the most obvious(P < 0.05).3.TUNEL staining: compared with the sham operation group,the neuronal apoptosis in the model group and solvent group increased significantly,and compared with the solvent group,the number of neuronal apoptosis in the model + oridonin group decreased significantly.4.Iba-1 immunofluorescence staining: compared with the sham operation group,the number of microglia in the model group and solvent group increased significantly,compared with the solvent group,the number of microglia in the model + oridonin medium dose group decreased significantly.5.Western blot results: compared with the sham operation group,the expressions of NLRP3 inflammatory bodies,ASC,caspas-1,IL-1 ?,IL-18 and apoptosis-related proteins Bax and caspase-3 in the model group and solvent group were higher than those in the solvent group.The expressions of NLRP3 inflammatory bodies,ASC,caspas-1,IL-1 ?,IL-18 and apoptosis-related proteins Bax and caspase-3 were significantly decreased in the model +oridonin group.Conclusion:oridonin can improve the early brain injury after SAH by reducing the formation of NLRP3 inflammasome,neuronal apoptosis and microglia activation.