| Objective: To investigate the effect of Shennaofuyuan decoction on the expression of P2X7 R and NLRP3 protein and the levels of Il-10,il-1β and Il-18 in the brain tissue after cerebral ischemia reperfusion in rats,to explore the possible mechanism of Shennaofuyuan decoction in reducing inflammatory reaction after cerebral ischemia reperfusion,and to provide scientific basis for clinical medication.Methods: 48 SD rats were randomly divided into four groups(Sham Operation Group,model group,traditional Chinese Medicine Group and positive medicine group)according to the method of random number table,after 2 hours,the suture emboli were pulled out gently to achieve cerebral ischemia and reperfusion,the Rats in the Chinese medicine group were given Shennaofuyuan decoction 8.7ml/kg by Gavage 5 days before the model was established,the rats in the positive medicine group were given BBG 30 mg/kg by intraperitoneal injection after 30 min of ischemia,and the rats in the other two groups were given the same amount of distilled water by Gavage and the same amount of physiological saline by intraperitoneal injection,after the rats were killed at different time(12h,24h),the expression of P2X7 R and NLRP3 mrna in the ischemic side of the brain was detected by RT-PCR,to investigate the effect of Shennaofuyuan decoction on inflammatory reaction in CIRI rats through P2X7R/NLRP3 pathway.Results:1.Neurologic impairment score: compared with the Sham Operation Group,the neurologic impairment score of the model group,the Shennaofuyuan decoction group and the BBG Group was significantly increased(P<0.01),and the difference was statistically significant Compared with the model group,the neurological deficit scores in the 12 h and 24 h Shennao Fuyuan decoction group and BBG Group were significantly decreased(P<0.05 or P<0.01),and there was no significant difference between the Shennao Fuyuan decoction group and BBG Group(P>0.05).2.HE staining: the morphology of the neurons in the ischemic side of the brain was observed under the microscope.The neurons in the hippocampus and cortex of the sham-operation group were well-defined and arranged with clear nucleoli and light nuclei.In the model group,the shape of nerve cells was not clear,the distribution was scattered,the cell body was reduced,the nucleolus was cleaved,the karyopyknosis was obvious,a lot of vacuoles were formed,and the staining was not even,especially at 12 h.Compared with the model group,the nerve cells in the shennao Fuyuan decoction group and BBG Group were less damaged,the cell body deformation was reduced,the number of vacuoles was reduced,the cell outline was clearer,and the arrangement of cells was relatively orderly in all phases(12h,24 h).3.The levels of IL-10,IL-1β and IL-18 in the brain tissue of rats: compared with the Sham Operation Group,the levels of IL-10 in the model group,the Shennaofuyuan Decoction Group and the BBG group were significantly increased(P<0.01),After intervention with Shennaofuyuan decoction and BBG,compared with the model group,IL-10 content in each phase(12h,24h)increased significantly(P<0.01),and there was no significant difference between the two intervention groups(P<0.05).The levels of IL-1β and IL-18 in model group,Shennaofuyuan decoction group and BBG Group were significantly higher than those in Sham Operation Group(P<0.01),the levels of IL-1β and IL-18 in BBG group were significantly decreased(P<0.01)at 12 h and 24 h,while the levels of IL-1β and IL-18 in TCM group were significantly decreased(P<0.05)at 12 h and 24 h,and there was no significant difference between the two intervention groups(P>0.05).4.The expression of P2X7 R and NLRP3 mrna in the ischemic brain tissue of rats: compared with the sham-operated group,the expression of P2X7 R and NLRP3 mrna in the model group,the Shennao Fuyuan decoction group and the BBG group were significantly increased(P<0.05 or P<0.01),the difference was statistically significant.Compared with the model group,the contents of P2X7 R and NLRP3 in the 12 h Shennaofuyuan decoction group were significantly lower(P<0.05),the contents of P2X7 R and NLRP3 in 24 h Shennaofuyuan decoction group and BBG Group at 12 h and 24 h decreased significantly(P<0.01).There was no significant difference between the two intervention groups(P>0.05).Conclusion: Shennaofuyuan decoction can effectively improve the neurological deficit after cerebral ischemia-reperfusion,reduce the inflammatory reaction and promote the recovery of neurological function.The mechanism of Shennaofuyuan decoction in treating ischemic stroke may be through inhibiting the expression of P2X7 R,decreasing the activation of NLRP3 inflammasome,decreasing the levels of IL-1β and IL-18 in brain tissue,increasing the levels of IL-10,and reducing the inflammatory reaction after infarction,to promote the recovery of neural function. |
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