| Obj ective:To observe the effects of acupuncture combined with rehabilitation method on NLRP3 inflammatory vesicles and Pyroptosis in neonatal neurons of hypoxic-ischemic brain injury,and to explore the possible mechanism of improving motor deficits after hypoxic-ischemic brain injury,in order to provide a relevant theoretical basis for the treatment of HIBD by acupuncture combined with rehabilitation method.Method:The 7-day-old rats were divided into control group(group A),model group(group B),acupuncture group(group C),rehabilitation group(group D),and acupuncture combined with rehabilitation method group(group E).Each group was assigned with 24h,7 days,and 14 days.HIBD animal model was established by improved Rice method.Group A and group B were not given intervention,group C was treated with head point cluster acupuncture,group E was treated with cage and rod rotation training,and group E was treated with cage and rod rotation on the basis of acupuncture.At each time point,motor impairment was evaluated by screen test,pyroptosis of nerve cells in hippocampal region was observed by TUNEL+caspase-1 fluorescence double-standard method,and NLRP3,caspase-1 and GSDMD-N protein expression in hippocampal region was detected by western blot.Results:1.Motor impairment score:Compared with group A,the screen test score of newborn rats in the other four groups was significantly higher at each time points,and there was a significant difference(P<0.05);Compared with group B,the experimental score of the therapy groups was significantly decreased at each time points,and there was a significant difference(P<0.05).Compared with groups C and D,the screen test scores of E group decreased at all each time points,and there was a significant difference(P<0.05).2.pyroptosis of hippocampal nerve cells:Except group A,the pyroptosis rate of hippocampal nerve cells in the other four groups was the highest 24 hours after operation,and the pyroptosis rate of nerve cells showed a significant decreasing trend with the extension of hypoxia and ischemia time(P<0.05);Compared with group A,the pyroptosis rate of hippocampal nerve cells in the other four groups was significantly up-regulated at each time points,and the difference was significant(P<0.05).Compared with group B,the pyroptosis rate of hippocampal nerve cells in all therapy groups decreased significantly at each time points,and the difference was significant(P<0.05).Compared with groups C and D,the pyroptosis rate of hippocampal nerve cells in group E was significantly decreased at each time points,and the difference was significant(P<0.05).3.NLRP3,caspase-1 and GSDMD-N expressions in hippocampus:Except for group A,the expressions of NLRP3,Caspase-1 and GSDMD-N proteins in the other four groups were the highest at 24h after surgery,and the expressions of NLRP3,Caspase-1 and GSDMD-N showed a decreasing trend with the extension of hypoxia and ischemia time.And its trend is consistent with the trend of pyroptosis of nerve cells.Compared with group A,the expressions of NLRP3,Caspase-1 and GSDMD-N in the other four groups were significantly increased at each time points,with statistical significance(P<0.05).Compared with group B,the expressions of NLRP3,Caspase-1 and GSDMD-N in each therapy group were significantly decreased at each time points,with statistical significance(P<0.05).Compared with C and D groups,NLRP3,Caspase-1 and GSDMD-N protein levels in E group were significantly decreased at each time points,and the differences were statistically significant(P<0.05).Conclusion:1.Acupuncture combined with rehabilitation Therapy can improve the motor function injury of newborn rats with HIBD,and is superior to single acupuncture or rehabilitation2.Acupuncture combined with rehabilitation Therapy can regulate the pyroptosis of nerve cells in hippocampal area of newborn rats with HIBD,and is superior to acupuncture or rehabilitation method.3.Acupuncture combined with rehabilitation method improves hypoxic-ischemic brain injury in neonatal rats by inhibiting NLRP3 inflammatory vesicle-mediated pyroptosis and regulating the cascade response to inflammation. |
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