| ObjectiveThis study intends to study the mechanism of mitochondrial damage in rat brain,heart,kidney and other related organ tissues under hypoxia,by using the method of morphology and molecular biology.To explore the molecular pathological mechanisms associated with mitochondrial dysfunction and cognitive decline.Based on the syndromes of traditional Chinese medicine,pathogenesis theory,overall concept and syndrome differentiation,combined with modern medical research,to explore the intervention effect and mechanism of Shenzhi Jiannao Prescription on mitochondrial damage in brain and related organ tissues of cognitive dysfunction rats.The pathological mechanism of mitochondrial function impairment provides new ideas,and the participation mechanism of the mitochondrial pathway in decreasing cognitive function provides experimental evidence for animals.Shenzhi Jiannao Prescription provides laboratory evidence and theoretical basis for clinical prevention and treatment of cognitive decline diseases.MethodsThe rats were randomly divided into normal group,model group and intervention group.An animal model of cognitive decline was established by hypoxia method.After the end of the experiment,the learning and memory abilities of the rats were tested using the Morris water maze behavioral test,and the rat model of cognitive decline was evaluated.The general state and behavioral test results were obtained.To observe the intervention effect of Shenzhi Jiannao Prescription on mitochondrial damage in rats with cognitive impairment.HE staining was used to observe the morphological changes of rat brain and other organ tissues;purified mitochondria of brain,heart,kidney and other organ tissues were extracted,and tissue mitochondrial respiratory chain enzyme complex I(NADH dehydrogenation)was detected by activity colorimetry.The total activity and specific activity of enzyme),mitochondrial respiratory chain enzyme complex Ⅳ(cytochrome C oxidase),the degree of mitochondrial swelling detected by spectrophotometry,and detection by enzyme-linked immunosorbent assay(ELISA)and immunohistochemistry The changes of mitochondrial cytochrome c(Cytochrome c,Cyt-c)expression were studied to clarify the mechanisms of hypoxia-induced impairment of mitochondrial damage in rats and the intervention of Shenzlii Jiannao Prescription.ResultsIn the first experiment,the results of behavioral experiments showed that in the comparative analysis of escape latency in each group,on the first day of the experiment,there was no significant change in the escape latency of the model group compared with the normal group and the intervention group(P>0.05,P>0.05).On the second day,compared with the normal group and the intervention group,the escape latency of the model group was prolonged,and the difference was statistically significant(P<0.01,P<0.01).On the third day,compared with the normal group and the intervention group,the escape latency of the model group was significantly prolonged,with a significant difference(P<0.01,P<0.01).On the fourth day,compared with the normal group and the intervention group,the escape latency of the model group was significantly prolonged(P<0.01,P<0.01).There was no significant difference in the escape latency between the normal group and the intervention group(P>0.05).The results of positioning navigation experiments showed that the total length of the swimming path of the rats compared with the normal group and the intervention group on the first day,there was no significant difference in the total length of the swimming path in the model group(P>0.05,P>0.05).On the second day,compared with the normal group and the intervention group,the total length of the swimming path in the model group increased significantly,and there was a significant difference(P<0.01,P<0.01).On the third day,compared with the normal group and the intervention group,the total length of swimming path in the model group increased significantly,and there was a significant difference(P<0.01,P<0.01).On the fourth day,compared with the normal group and the intervention group,the total length of the swimming path in the model group increased significantly(P<0.01,P<0.01).Compared with the normal group,the total length of the swimming path in the intervention group had no Significant differences(P>0.05).Space exploration experiments showed that the ratio of swimming time and total swimming time in the original platform quadrant was significantly lower in the model group than in the normal group and the intervention group(P<0.01,P<0.05).There was no significant difference between the normal group and the intervention group.The number of rats crossing the target platform was significantly reduced in the model group compared with the normal group and the intervention group(P<0.01,P<0.01).There was no significant difference between the normal group and the intervention group.In the second experiment,morphological results showed that compared with the normal group,the morphological results of brain and related organ tissues in the model group showed obvious pathological changes,with loosely arranged cells,widened gaps,blurred cell outlines,unclear boundaries,and a few nucleus.Pyknosis,dissolution,necrosis,and capillary congestion,dilation,hemorrhage,inflammatory cell infiltration and other diseases.Compared with the normal group,the degree of injury in the intervention group was significantly lower than that in the model group.In the third experiment,the results showed that the specific activities of mitochondrial respiratory chain enzyme complexes I and IV in rat brain were significantly lower in the model group than in the normal group(P<0.01,P<0.05),and compared with the intervention group.In contrast,the model group was significantly lower than the intervention group(P<0.01,P<0.05);there was no significant difference between the normal group and the intervention group.The specific activities of mitochondrial respiratory chain enzyme I and IV in rat cardiac tissue were significantly lower in the model group than in the normal group(P<0.01,P<0.01),and significantly lower in the model group than in the intervention group.In the intervention group(P<0.01,P<0.01),there was no significant difference between the normal group and the intervention group.The specific activity of mitochondrial respiratory chain enzyme complexes I and IV in rat kidney tissue was significantly lower in the model group than in the normal group(P<0.01,P<0.01);the model group was significantly lower than the intervention group.In the intervention group(P<0.01,P<0.01),there was no significant difference between the normal group and the intervention group.In the fourth experiment,the results showed that the degree of mitochondrial swelling in rat brain,heart,and kidney tissues was significantly higher in the model group than in the normal group(P<0.05,P<0.01,P<0.05);The ratio of mitochondrial swelling in the model group was significantly higher(P<0.05,P<0.01,P<0.05).There was no significant difference between the normal group and the intervention group.The expression of mitochondrial Cytc in rat brain,heart and kidney tissues was significantly higher in the model group than in the normal group(P<0.01,P<0.01,P<0.01).Compared with the intervention group,the model group was significantly higher(P<0.01,P<0.05,P<0.01);There was no significant difference between the normal group and the intervention group.The results of immunohistochemistry showed that compared with the normal group,the positive expression in the model group was significantly higher than that in the normal group(P<0.05,P<0.05,P<0.05);compared with the intervention group,the model group was significantly higher than the model group(P<0.05,P<0.05,P<0.05);there was no significant difference between the normal group and the intervention group.ConclusionHypoxic method replicates a cognitive decline model in rats and influences learning and memory related cognitive functions in rats,which is consistent with the performance of cognitive decline models in rats,suggesting the feasibility of hypoxic modelling and animal models The establishment of.The results of behavioral experiments,morphological and molecular biological methods show that mitochondrial dysfunction is closely related to the pathological mechanism of cognitive decline.Based on the pathogenesis of traditional Chinese medicine and disease syndrome theory,the overall concept,syndrome differentiation and treatment as a guide,the brain The method can prevent and protect cognitive function through mitochondrial pathway,and effecti vely improve the related symptoms of cognitive dysfunction rats. |
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