Based On The Theory Of "the Spleen Stores Intention And The Kidney Stores Will", The Mechanism Of Action Of Tonifying Spleen And Kidney On Rats With Cognitive Dysfunction Model Was Explore

Objective:To systematically discuss the relationship between cognitive dysfunction and"the spleen stores meaning and the kidney stores aspirations"with the theoretical guidance of"spleen stores meaning,kidney stores aspirations",and researches the prescription of modern Chinese medicine for treating dementia based on the ancient and modern medical record cloud platform The regularity of medication,and the use of Bupi Yishen prescription to intervene in cognitive dysfunction model rats,by detecting the levels of TNF-α,IL-1β,IL-6,and IFN-γin the serum and brain tissue of the rats,it was verified that tonifying the spleen and nourishing the kidney The anti-inflammatory effect of the prescription on cognitive dysfunction rats;the detection of M1/M2 microglia markers in brain tissue,and the investigation of the activation and polarization of Bupi Yishen prescription on microglial cells;detection of TLR4/MYD88/The expression levels of key targets of NF-κB signaling pathway elucidate the mechanism by which Bupi Yishen Decoction inhibits the activation of microglia by inhibiting the over-activation of TLR4/MYD88/NF-κB signaling pathway,thereby achieving anti-inflammatory effect.Material and method:Part 1:Theoretical PartThrough the collection and analysis of ancient Chinese medicine books and documents,starting from the ancient understanding and naming of cognitive dysfunction,as well as the understanding of etiology and pathogenesis,to sort out the relationship between"spleen stores meaning,kidney stores wisdom"and cognitive dysfunction,Further explain the advantages of using the method of"invigorating the spleen and nourishing the kidney"and traditional Chinese medicine to prevent and treat cognitive dysfunction.Part II:Data MiningThrough computer retrieval of valid cases of modern literature on dementia in databases such as CNKI,WANFANG DATE,VIP,etc.,data screening,data cleaning,data standardization,etc.Using the ancient and modern medical record cloud platform to carry out data analysis methods such as frequency statistics,association analysis,cluster analysis,etc.,and conduct complex network analysis and visualized display,analyze the frequency of use of traditional Chinese medicine in the treatment of dementia,the four qi,five flavors,meridians,core prescriptions and other medication rules.Part III:Experimental StudySixty-two SPF SD rats were randomly divided into two groups:13 in the sham operation group and 49 in the modeling group.The rat model of cognitive dysfunction was replicated by injecting Aβ1-42 into the lateral ventricle.After modeling,a water maze and open field test were performed to evaluate the cognitive function of the two groups of rats.One of the rats was randomly selected for HE staining to observe the pathological changes of the rat brain tissue.Then the remaining 48 rats in the modeling group were randomly divided into four groups:model control group,high-dose Bupi Yishen prescription group,Bupi Yishen prescription medium-dose group,and Bupi Yishen prescription low-dose group.12.The rats in the sham operation group were given normal saline by oral gavage,once a day,for 6consecutive weeks;the rats in the high-dose,medium-and low-dose Bupi Yishen Fang groups were given high,medium and low concentrations of Bupi Yishen Decoction,respectively.The rats in the model control group were given the same amount of normal saline by oral gavage,once a day for 6 consecutive weeks.The water maze training was started 6 days before the end of the administration,once a day for 5 consecutive days,and the water maze test was performed on the sixth day;1 day before the end of the administration,the rats in each group underwent an open field test;2 hours after the last administration,3%sodium pentobarbital（1.5ml/kg）was injected intraperitoneally,whole blood was collected by abdominal aorta puncture,and the contents of TNF-α,IL-1β,IL-6 and IFN-γin serum were detected by ELISA;6 rats were randomly selected from the group,the whole brains were taken,the contents of TNF-α,IL-1β,IL-6 and IFN-γin the cortex were detected by ELISA,and Aβ1-42 in hippocampus and Aβ1-42 in the cortex were detected by western-blot method.The expression levels of TLR4,MYD88,NF-κB and p-NF-κB;the remaining 6 rats in each group were taken as whole brains,immersed in 4%paraformaldehyde solution for fixation,and HE staining was used to observe the brains of rats in each group Histopathological changes;immunofluorescence double-labeling method was used to detect Co-expression of Iba-1/CD80（B7-1）,Iba-1/CD86（B7-2）,Iba-1/CD206,Iba-1/Arg-1,Iba-1/p-N F-κB.Results:Part 1:Theoretical PartCognitive dysfunction belongs to the category of diseases such as"dementia","forgetfulness"and"foolishness"in traditional Chinese medicine.The"Nei Jing"has the theory of"five zang organs and spirits".The medical practitioners of all dynasties took the"Nei Jing"as its purpose and advocated that consciousness,thinking,emotions and other mental activities are dominated by the heart and brain,and the five zang organs are closely related.For cognitive function,the focus is on the theory that"the spleen stores meaning and the kidney stores mind".The kidney is the innate and the spleen is the acquired."The spleen stores the camp,and the camp houses the meaning;the kidney stores the essence,and the essence houses the will."The spleen is in charge of transporting and transforming the essence of water and grain,and the kidney is in charge of storing the essence.It is the material basis of cognitive function,and it is embodied in memory,calculation,language,visual space,and understanding and judgment.Diseases such as"dementia","forgetfulness",and"foolishness"are characterized by a lack of healthy functioning of the spleen,deficiency of the spleen and production of phlegm;deficiency of kidney essence and atrophy of the brain and marrow.Decreased ability;severe dementia,language disorders,visual space abnormalities,mental disorders,etc.The main treatment is to invigorate qi and spleen,invigorate kidney and brain,clear phlegm and open orifices.Part II:Data MiningA total of 144 literatures were included,including 154 prescriptions,among which the core drugs were Shichangpu,Chuanxiong,Polygala Radix,Astragalus,Shudi,Salvia,etc.Most of the drugs were Ganping,Xinwen,Bitterhan,and the drugs entered the heart,liver,and spleen in turn.,Kidney meridian.The clustering results include 7 categories,and the drug combinations with high frequency include Shichangpu-Ganchi,Astragalus-Salvia,Poria-Glycyrrhiza,etc.Part III:Experimental StudyThe results of the water maze test showed that through the water maze practice,the positioning and navigation time of the rats in each group was gradually shortened,and the escape latency of the rats in the sham operation group decreased the fastest.The results of the last positioning navigation and space exploration experiments showed that compared with the sham-operated group,the escape latency of rats in the other groups increased significantly（p<0.01）,and the number of crossing platforms was significantly decreased（p<0.01）,and the differences were statistically significant;Compared with the model control group,the escape latency of the rats in each dose group of Bupi Yishen Recipe was significantly decreased（p<0.01）,and the number of crossing platforms was significantly increased（p<0.01）,especially in the high-dose Bupi Yishen Recipe group.obvious.The results of the open field test showed that compared with the sham-operated group,the rats in the model control group and each dose group of Bupi Yishen prescription were significantly reduced in the number of times of standing upright in the dark room and the number of times of washing their faces（p<0.05）,and the difference was statistically significant.Significant;compared with the model control group,the number of times of standing upright in the dark room and the number of times of washing the face of the rats in each dose group of Bupi Yishen Recipe were significantly increased（p<0.05）,and the difference was statistically significant,especially in the high dose group.for obvious.The detection results of TNF-α,IL-1β,IL-6 and IFN-γin serum and cerebral cortex showed that compared with the sham operation group,the serum of rats in the model control group and the high,medium and low dose groups of Bupi Yishen prescription The contents of TNF-α,IL-1β,IL-6 and IFN-γin cerebral cortex were significantly increased（p<0.01）,and the difference was statistically significant;The contents of TNF-α,IL-1β,IL-6and IFN-γin the serum and cerebral cortex of the rats in the low-dose group were significantly decreased（p<0.05）,and the difference was statistically significant.Western-blot method was used to detect the expression levels of Aβ^1-42in hippocampus and TLR4,MYD88,NF-κB and p-NF-κB in cerebral cortex.The expression levels of Aβ1-42 in the hippocampus and TLR4,MYD88,NF-κB and p-NF-κB in the cerebral cortex of the rats in the low-dose group were significantly increased（p<0.01）,and the difference was statistically significant;In comparison,the expression levels of Aβ1-42 in the hippocampus and TLR4,MYD88,NF-κB,and p-NF-κB in the cerebral cortex of the rats in the high,medium and low dose Bupi Yishen prescription groups were significantly decreased（p<0.05）.statistically significant.The results of immunofluorescence detection showed that compared with the sham operation group,the levels of Iba-1/CD80（B7-1）and Iba-1/CD86（B7-2）and Iba-1/p-NF-κB double-positive cells were significantly increased（p<0.01）,and Iba-1/CD206,Iba-1/Arg-1double-positive cells were significantly decreased（p<0.05）,the difference was statistically significant;compared with the model control group,the levels of Iba-1/CD80（B7-1）,Iba-1/CD86（B7-2）,the number of Iba-1/p-NF-κB double-positive cells decreased significantly（p<0.05）,and the double-positive cells of Iba-1/CD206 and Iba-1/Arg-1 were significantly increased（p<0.05）.Statistical significance.Conclusion:1.Deficiency of spleen and kidney,empty marrow and phlegm coagulation are the key pathogenesis of cognitive dysfunction.Based on the theory of"spleen stores meaning and kidney stores mind",through the treatment of spleen and kidney,it is possible to replenish the brain and marrow,resolve phlegm and open orifices.Make the will accessible and the spirit healed.2.Through the method of data mining,it is concluded that the core drugs of traditional Chinese medicine for the treatment of dementia are Shichangpu,Chuanxiong,Polygala,Astragalus,Shudi,etc.Most of the drugs are Ganping,Xinwen,Bitter Cold,and the drugs enter the heart,liver,The spleen and kidney meridians;the frequently used drug combinations include Shichangpu-Polygala,etc.;with the main effect of opening the orifices,and taking into account the regulation of qi at the same time,it has a therapeutic effect on dementia from the two aspects of the specimen.3.Bupi Yishen Decoction can improve cognitive function in rats with cognitive dysfunction,reduce the level of inflammatory factors,inhibit the over-activation of M1-type microglia,and promote the polarization of microglia to M2-type.It was achieved by inhibiting the activation of TLR4/MYD88/NF-κB signaling pathway in microglia.