| Purpose:Under the guidance of the theory of“spleen brain correlation”,this paper systematically discusses the correlation between cognitive impairment and spleen,and establishes a cognitive impairment due to spleen deficiency by combining disease with syndrome,so as to explore the curative effect and mechanism of Bupi Xingshen Yizhi prescription on cognitive impairment due to spleen deficiency,and to provide theoretical and experimental basis for clinical prevention and treatment of cognitive impairment.Material and method:Part one:theoretical researchThrough the collation of ancient literature and the analysis of related theories,this paper combs the disease name,etiology and pathogenesis of cognitive dysfunction,discusses the close relationship between spleen and cognitive dysfunction from spleen and brain,spleen and cognitive function,spleen and aging,and explores the advantages and broad application prospects of invigorating spleen,arousing mind and replenishing wisdom as an important treatment for preventing and treating cognitive impairment.Part two:experimental study60 SPF SD rats,half male and half female,were randomly divided into 5 groups:normal group,cognitive impairment group,spleen deficiency cognitive impairment group,Aricept group,Bupi Xingshen Yizhi formula group,12 rats in each group,adaptive feeding for one week.The cognitive impairment model of spleen deficiency was established by complex factors,combination of disease and syndrome and hippocampal injection of Aβ1-42.In the second and third weeks,the model of spleen deficiency was established.In the fourth week,Aβ1-42 was injected into hippocampal CA1 area of cognitive impairment group,spleen deficiency cognitive impairment group,Aricept group,Bupi Xingshen Yizhi formula group.The normal group,cognitive impairment group and spleen deficiency cognitive impairment group were given 0.9%normal saline by gavage during treatment,and the remaining two groups were given corresponding drugs by gavage.The treatment cycle was 4 weeks.The water maze adaptive training,positioning navigation and space exploration experiments were carried out on the fourth week of gastric perfusion in order to evaluate the learning and memory ability of rats among different groups.After the last gastric perfusion,the rats were anesthetized for 24 hours after fasting.The morphological changes of neurons in brain tissue were observed and analyzed by HE staining.The expression of serum TNF-α,IL-1βand IL-6 were detected by ELISA.The mRNA and protein expressions of Aβ,NF-κB,TREM2,DAP12,IκB and ERK1/2 in brain tissue were detected by RT-PCR and WB method,and the possible mechanism of Bupi Xingshen Yizhi recipe in improving learning and memory impairment of rats with spleen deficiency cognitive impairment was observed and discussed from the perspective of inflammatory reaction.Results:Part one:theoretical research1 Based on the analysis of the related contents of cognitive impairment in ancient books,the descriptions of the related diseases are mainly "forgetting" and "staying",and show a progressive relationship in the severity of the disease.2 Spleen deficiency is a common syndrome in elderly people and patients with cognitive impairment Spleen is connected with brain meridians and Qi and blood are interlinked.Spleen deficiency leads to brain marrow dystrophy,spleen dyskinesia leads to phlegm and turbid endogenesis.Deficiency leads to blood stasis,phlegm and blood stasis obstructs orifices,which leads to cognitive dysfunction.Therefore,the prevention and treatment of cognitive dysfunction can be based on the theory of "spleen and brain”.3 The method of invigorating the spleen and arousing the mind and replenishing intelligence can fundamentally block the endogenesis of phlegm and blood stasis by invigorating the spleen and replenishing qi and blood,which is an effective method to prevent and treat cognitive dysfunction.Part two:experimental study1 General state of rats results:after the treatment,compared with the normal group,the cognitive impairment group and the spleen deficiency cognitive impairment group showed different degrees of mental fatigue,rough hair,lack of luster,loose stool,accompanied by squint and drowsiness.Compared with the two model groups,the general state of each treatment group was improved,and the improvement degree of Bupi Xingshen Yizhi formula group was better than that of anlishen group.2 Results of behavioral experiments2.1 Place navigation:compared with the normal group,the escape latency of cognitive impairment group and spleen deficiency cognitive impairment group was significantly increased(P<0.01),compared with the two model groups,the escape latency of each treatment group was significantly improved(P<0.01),and there was no significant difference between the two treatment groups(P>0.05).2.2 Space exploration experiment:compared with the normal group,the time required to cross the target area for the first time in the cognitive impairment group and the spleen deficiency cognitive impairment group was significantly longer,the number of times to cross the area was significantly reduced,and the residence time in the target quadrant was shortened(P<0.01).The target quadrant time in the spleen deficiency cognitive impairment group was longer than that in the cognitive impairment group(P<0.05).Compared with the model group,the time required to cross the target area for the first time,the number of crossing and the residence time of the target quadrant in each treatment group were better than those in the model group.In the space exploration experiment,the normal group’s trajectory is relatively simple,and there are repeated reentry exploration behavior.The dementia group and the spleen deficiency dementia group’s overall trajectory is chaotic,and the residence time in each quadrant is relatively balanced.3 Morphological results:compared with the normal group,the number of neurons in the hippocampus and the cortex around the hippocampus in the cognitive impairment group and the spleen deficiency cognitive impairment group was significantly reduced,the arrangement was sparse,the cell volume was increased,the nucleolus pyknosis,the nuclear staining was deepened,the number of pyramidal cells was reduced,the arrangement was disordered,and vacuolar degeneration appeared.Compared with the model group,the number of neurons in the hippocampus and perihippocampal cortex in anlishen group and Bupi Xingshen Yizhi formula group increased significantly,but it was still lower than that in the normal group.The cell morphology was more regular,the number of dendrites in pyramidal cells increased significantly,and the number of vacuolar degeneration and cell pyknosis and necrosis decreased.4 ELISA results:compared with the normal group,the expression levels of IL-1β,IL-6 and TNF-α in cognitive impairment group and spleen deficiency cognitive impairment group were significantly increased(P<0.01),the increase range of spleen deficiency cognitive impairment group was significantly higher than that of cognitive impairment group.The expression levels of IL-1β,IL-6 and TNF-α in Aricept group and Bupi Xingshen Yizhi formula group were significantly decreased compared with two model groups,the down-regulation effect of Bupi Xingshen Yizhi formula group was more significant5 RT-PCR results:compared with the normal group,the relative expression of NF-κB in cognitive impairment group and spleen deficiency cognitive impairment group was significantly up-regulated(P<0.01),the relative expression of IκB was down regulated(P<0.01).Compared with the cognitive impairment group,the expression of NF-κB mRNA in the two treatment groups were significantly down regulated,the relative expression of IκB was up-regulated.Compared with the normal group,the relative expression of TREM2 mRNA and DAP12 mRNA in cognitive impairment group and spleen deficiency cognitive impairment group were significantly down regulated(P<0.01).Compared with the two model groups,the relative expression of TREM2 mRNA in Aricept group and Bupi Xingshen Yizhi formula group were up-regulated(P<0.01),and the up-regulation effect of Bupi Xingshen Yizhi formula was better than that of Aricept group(P<0.01).Compared with the normal group,the relative expression of ERK1 and ERK2 mRNA in the cognitive impairment group and the spleen deficiency cognitive impairment group were significantly increased(P<0.01),compared with the two model groups,the relative expression of ERK1 and ERK2 mRNA were significantly decreased,but there was no statistical significance between the two treatment groups.6 Western blot results:compared with the normal group,the protein expression levels of Aβ,NF-κB and ERK1/2 in hippocampus of cognitive impairment group and spleen deficiency cognitive impairment group were significantly increased(P<0.01),and the protein expression levels of IκB,TREM2 and DAP 12 were significantly decreased(P<0.01).Compared with the spleen deficiency cognitive impairment group,the protein expression levels of Aβ,NF-κB and ERK1/2 in Aricept group and Bupi Xingshen Yizhi Fang group were significantly decreased(P<0.01).The protein expression levels of IκB TREM2 and DAP12 were significantly increased(P<0.01).In addition,the up-regulation range of TREM2 and down-regulation range of Aβ in Bupi Xingshen Yizhi Decoction group were significantly higher than Aricept group,the difference was statistically significant(P<0.05).Conclusion:1 Spleen deficiency is the key pathogenesis of cognitive impairment,and is closely related to the main pathological products of cognitive impairment,such as phlegm and blood stasis.Based on the theory of "spleen brain correlation",the method of invigorating the spleen and invigorating the mind and replenishing intelligence can promote the movement of Qi and blood,and then nourish the brain,harmonize qi and blood,awaken the mind and open the orifices.It has sufficient theoretical basis and wide application prospects for the prevention and treatment of cognitive dysfunction.2 Spleen deficiency complex factors modeling method combined with Aβ1-42 hippocampal injection can successfully construct the spleen deficiency cognitive impairment model,which can lead to learning and memory impairment and inflammatory damage,and more appropriately copy the clinical symptoms of patients with cognitive impairment3 Bupi Xingshen Yizhi recipe can significantly improve the symptoms of spleen deficiency syndrome,such as loose stool,rough hair,mental depression,etc.in rats with cognitive impairment due to spleen deficiency,at the same time,it can improve the positioning cruise and spatial exploration ability of model rats,and restore their learning and memory ability.4 The mechanism of Bupi Xingshen Yizhi recipe in improving cognitive function is related to TREM2/NF-κB signaling pathway.It can prevent and treat cognitive dysfunction by up regulating the mRNA and protein expression of TREM2 and its ligand DAP 12,inhibiting the activation of NF-κB inflammatory pathway and reducing the content of Aβ and inflammatory factors in brain tissue. |
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