| Objective Ischemic cerebrovascular disease(stroke)has become a problem of medical intervention and rehabilitation due to its high incidence,high disability rate and high recurrence rate.The existing clinical prevention and treatment measures are mainly early detection,ultra-early thrombolysis,anticoagulation,but their effects are often unsatisfactory.Finding new prevention methods and theoretical exploration has become a top priority.The rise of the “stroke unit” in the past 20 years is the most effective treatment and rehabilitation method for stroke.However,because the recurrence rate and disability rate of stroke are still high,it is particularly important to find more effective methods to prevent ischemic stroke.Hyperbaric oxygen intervention can increase the permeability of oxygen and other substances,directly supply oxygen to hypoxic nerve cells,relieve cell edema,restore cell function,and thus get more and more rescue in the acute phase of stroke.The Micro-Hyperbaric Oxygen Aromatherapy technology improves the high-pressure oxygen system,that is,the micro-pressurization of the cabin pressure,and the oxygen separation technology is established by molecular sieve air separation to establish an environment higher than the standard atmospheric pressure of 20 k Pa-30 k Pa.Molecular sieve technology produces oxygen concentration of up to 93% ± 3%(medical grade),this oxygen concentration is diffused into the tank,so that the concentration in the oxygen chamber is maintained at 25% under the pressure of 20 k Pa-30 k Pa."Aromatic" is an aromatic active small molecule substance,which is highly purified by distillation or extraction technology,making it a substance that can directly absorb the odor through the olfactory pathway(essential oil).The Micro-Hyperbaric Oxygen Aromatherapy system combines the three to form a multi-effect superposition of oxygen therapy and aromatherapy in a micro-pressure space,which not only maintains the effectiveness of hyperbaric oxygen,but also uses aromatic small molecules as an important auxiliary means.It is used to form a new intervention for stroke.The MCAO rat model is a classic model for studying ischemic stroke.As a method to simulate human ischemic stroke,it is ideal for the study of stroke prevention,treatment,rehabilitation and mechanism.Therefore,the successful preparation of the MCAO model is the key to the smooth progress of the experiment.The laboratory has been engaged in a combination of aromatherapy,oxygen therapy and micro-pressure therapy and medical practice.In recent years,the Micro-Hyperbaric Oxygen Aromatherapy system has been established.In the early stage,we have studied the intervention of Micro-Hyperbaric Oxygen Aromatherapy after cerebral ischemia-reperfusion injury in rats.This experiment observes the research from the perspective of pretreatment,on the one hand to explore its mechanism,and on the other hand to explore diseases that threaten human health.Whether there is a preventive effect.Therefore,the experiment is divided into four parts: the first part of the preparation of MCAO model;the second part of MCAO rats neurobehavioral score;the third part of MCAO rats brain neuron morphology observation;the fourth part of immunohistochemistry and Western blot are used to detect protein expression.To explore whether Micro-Hyperbaric Oxygen Aromatherapy has an effect on ischemic stroke and its possible mechanism,and provide reference value for prevention of ischemic stroke.Part One Model preparation and neurological function scores of MCAO ratsMethods The Micro-Hyperbaric Oxygen Aromatherapy group and Micro-Hyperbaric Oxygen group were intervened 14 days before surgery,and the other groups were not treated with intervention.The cerebral ischemia-reperfusion injury model was established by blocking the middle cerebral artery with the suture method.After 2 h of ischemia,the plug was pulled out.Perfusion,the rats were scored with a Longa score.Rats scored 0 and 4 were excluded.Results The sham group had a neurological score of 0,and there was no abnormal behavior.The model group,Micro-Hyperbaric Oxygen group,Micro-Hyperbaric Oxygen Aromatherapy group scored from 1 to 3 points.The rats had obvious behaviors such as contralateral forepaw adduction,eye Horner sign,circle rotation and dumping.Part Two The observation of behavior of MCAO rats by MicroHyperbaric Oxygen AromatherapyMethods Neurological function scores,Berderson scores,and balance beam experiments were performed at 24 h after ischemia-reperfusion.The behavioral scores were partially sacrificed after completion of the behavioral scores.Brain sections were stained with TTC to calculate the percentage of cerebral infarction volume.Results The reperfusion of the 24 h Micro-Hyperbaric Oxygen Aromatherapy group was significantly different from the model group(P<0.05);the Micro-Hyperbaric Oxygen group and Micro-Hyperbaric Oxygen Aromatherapy group was statistically significant(P<0.05).Berderson score was significantly different between the sham group and the model group at 24 h after reperfusion(P<0.05).The difference between the Micro-Hyperbaric Oxygen Aromatherapy group and the model group was statistically significant(P<0.05);the Micro-Hyperbaric Oxygen group and Micro-Hyperbaric Oxygen Aromatherapy group was statistically significant(P<0.05).The results of TTC staining showed that there was different degree in model group and the Micro-Hyperbaric Oxygen Aromatherapy group.Infarct size,compared with the model group,the infarct volume of the Micro-Hyperbaric Oxygen Aromatherapy group was significantly reduced,and the difference was significant(P<0.05);compared with the Micro-Hyperbaric Oxygen group,the infarct volume of the Micro-Hyperbaric Oxygen Aromatherapy group was reduced(P<0.05).Part Three Observation of the morphology of nerve cells in MCAO rats by Micro-Hyperbaric Oxygen AromatherapyMethods Neurobehavioral experiments showed that Micro-Hyperbaric Oxygen Aromatherapy had protective effects on the neurological function of rats with cerebral ischemia-reperfusion injury.In order to explore the mechanism of Micro-Hyperbaric Oxygen Aromatherapy,some rats were sacrificed after the behavioral score was completed.The brain was taken after peracetal infusion,and the brain tissue of the pear-shaped cortex was cut through continuous brain tissue sections.HE staining was used to observe the morphology of nerve cells.Nissl staining was used to observe changes in the number of Nissl bodies in nerve cells.Results The results of HE staining showed that the sham group had normal morphology,the cell body was round and the nucleolus was obvious,and the cells were arranged in multiple layers,neat and dense.The number of cells in the model group was reduced,the arrangement was sparse,the cell body was shrunk,and the nucleus was solid.The triangle was reduced and some of the nuclei disappeared.The results of the Micro-Hyperbaric Oxygen Aromatherapy group and the Micro-Hyperbaric Oxygen group were improved to some extent compared with the model group.A few normal nerve cells were seen,and the arrangement was not neat.The results of Nissl staining showed that there were more dark blue plaque-like Nissl bodies in the cytoplasm of the sham rats.The coloration was deep and uniform,the cell morphology was normal,and the cells were arranged neatly.The model group had pear-shaped cortical nerve cells.The number of Nissl bodies in the cytoplasm was significantly reduced,and some even disappeared.The cell bodies were shrunk and the arrangement was sparse.Compared with the model group,the cytoplasm of the piriform cortical neurons in the Micro-Hyperbaric Oxygen Aromatherapy group and the Micro-Hyperbaric Oxygen group were relatively increased.Part Four The effect of Micro-Hyperbaric Oxygen Aromatherapy on the expression of Apaf-1/Caspase-3 in the brain of MCAO ratsMethods In order to further explore the mechanism of Micro-Hyperbaric Oxygen Aromatherapy,the expression of Apaf-1 protein in the pear-shaped cortex was detected by immunohistochemistry,the change of Apaf-1 protein and Caspase-3 protein content in the pear-shaped cortex was detected by Western blot.Results The results of immunohistochemistry showed that the positive expression of Apaf-1 protein was brownish yellow in the cytoplasm of nerve cells.In the sham group,the piriform cortical neurons of the rat brain were arranged neatly and in a normal shape with a small amount of cytoplasmic yellow staining.The morphology of the piriform cortical neurons in the Micro-Hyperbaric Oxygen Aromatherapy group and the Micro-Hyperbaric Oxygen group were significantly improved compared with the model group,with slight nuclear pyknosis and yellow staining of the cytoplasm of the nerve cells.Compared with the sham group,the number of Apaf-1 protein positive cells in the model group increased(P<0.05).Compared with the model group,the number of positive cells in the Micro-Hyperbaric Oxygen Aromatherapy group and the Micro-Hyperbaric Oxygen group were decreased(P<0.05).The results of Western blot showed that the average gray value of Apaf-1 protein and Caspase-3 protein in the model group were higher than that in the sham group(P<0.05).Compared with the model group,the Apaf-1 and Caspase-3 of the Micro-Hyperbaric Oxygen Aromatherapy group and the Micro-Hyperbaric Oxygen group were significantly decreased(P<0.05).Conclusion 1.Micro-Hyperbaric Oxygen Aromatherapy can produce neuroprotective effects on cerebral ischemia-reperfusion injury in rats.2.The mechanism of Micro-Hyperbaric Oxygen Aromatherapy is related to the decrease of Apaf-1 and Caspase-3 protein expression in brain Piriform Cortex.3.The combination of Micro-Hyperbaric Oxygen and aromatherapy produces a synergistic effect. |
PDF Full Text Request