| Purpose:To observe the levels of P2X7 R,NLRP3,pro-caspase-1,ASC,caspase-1,IL-1? and IL-18 expression in brain tissue of CIRI rats,and to explore whether the anti-inflammatory mechanism of the eye against CIRI rats is achieved by regulating cell death.Methods:85 SD rats were randomly divided into the Sham-operated group(16 rats)and the model group(69 rats);The CIRI model was prepared by the model of the rats in the model group.The rats in the sham operation group were sutured only after the neck muscle injury occurred in the rat neck.After successful modeling,rats were evaluated for behavioral signs and TTC staining to observe hypoxia in rat brain tissue.the model rats were randomly divided into model control group,eye-acupuncture group and inhibitor group.Rats in the sham-operated group were normally reared until the end of the experiment,and the model control rats were reared after reperfusion.The intervention time of the eye-acupuncture group was immediate reperfusion,8h,16 h,24h;Inhibitor group rats injected with Glibenclamide.After the last acupuncture,the general state of the rats was observed,and the behavioral signs were scored for each group of rats.The rats were sacrificed by excessive anaerobic chloral hydrate,and the morphological changes of the cells were observed by electron microscopy.ELISA was used to detect the contents of IL-1? and IL-18 in brain tissues.The expression levels of P2X7 R,NLRP3,ASC,pro-caspase-1and caspase-1 in brain tissues were detected by the methods of immunochemistry and western-blot.Results: 1.Effects of 1 eye on behavior score of CIRI ratsCompared with the sham operation group,the scores of behavioral signs of rats in the model control group increased significantly,and the difference was statistically significant(p < 0.01).Compared with the model control group,the scores of behavioral signs of rats in the eye acupuncture group and the inhibitor group were significantly increased,and the difference was statistically significant(p<0.01).2.The effect of the eye on the morphology of neuronsIn the sham operation group,neurons were intact,cell permeability was not abnormal,cell membrane integrity was not abnormal,and intracellular organelles and cell contents were not abnormal.In the model control group,the cells were swollen,the cell membrane was damaged,the integrity was destroyed,the cell contents and organelles were destroyed and released to the outside of the cell,and the cells were damaged.Eye acupuncture group: compared with the sham operation group,the rat neurons in the eye acupuncture group showed increased permeability,cell swelling,and occasional damage to intracellular organelles and contents.In the inhibitor group,the morphology of rat neurons was similar to that in the eye acupuncture group,with slightly increased membrane permeability,relatively swollen cells,but complete contents,and occasionally damaged organelles.3.The effect of eye on the expression levels of il-1 and il-18 inflammatory factors in brain tissues of CIRI ratsCompared with the sham operation group,the serum expression levels of il-1 and il-18 in the model control group were significantly increased.Compared with the model control group,the expression levels of il-1 and il-18 in brain tissues of rats in the ophthalmic acupuncture group and the inhibitor group were significantly reduced,and the difference was statistically significant(p <0.01).4The ophthalmic needle regulated the expression of pyroptosis related proteins in CIRI rats 4.1 Expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 in brain tissues were detected by western-blot method:Compared with the sham operation group,the expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the model control group were significantly enhanced,with statistically significant differences(p<0.01).Compared with the model control group,the expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 were significantly reduced in the eye acupuncture group(p<0.01).The expression levels of NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the inhibitor group were significantly reduced,and the difference was statistically significant(p<0.01).There was no significant change in the expression level of P2X7 R,and the difference was not statistically significant(p>0.05).Compared with the ophthalmic acupuncture group,the expression levels of NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the inhibitor group showed no significant change(p>0.05),and the expression level of P2X7 R increased significantly(p<0.01).4.2 The expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 in brain tissues were detected by immunohisochemistry:Compared with the sham operation group,the expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the model control group were significantly enhanced,with statistically significant differences(p<0.01).Compared with the model control group,the expression levels of P2X7 R,NLRP3,pro-caspase-1,ASC and caspase-1 were significantly reduced in the eye acupuncture group(p<0.01).The expression levels of NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the inhibitor group were significantly reduced,and the difference was statistically significant(p<0.01).Compared with the ophthalmic acupuncture group,the expression levels of NLRP3,pro-caspase-1,ASC and caspase-1 in the brain tissues of rats in the inhibitor group showed no significant change(p>0.05),and the expression level of P2X7 R increased significantly(p<0.01).Conclusion: 1.Eye acupuncture therapy can inhibit the occurrence of pyroptosis 2.Eye acupuncture therapy can inhibit the occurrence of pyroptosis by reducingthe expression of P2X7R?NLRP3?pro-caspase-1?ASC?caspase-1?IL-1??IL-18 and and play an anti-inflammatory role. |
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