Analysis Of Syndrome Elements Of Patients With Cognitive Impairment Based On The Pathogenesis Of Marrow Deficiency And Toxin Damage And Research On The Protective Mechanism Of Jiedu-Yizhi Formula On Aβ_25-35-induced AD Rats

Objective: Clinical part: To screen the elderly patients with cognitive impairment in some areas of Changchun,record their general information,disease history,relevant examination results,evaluate the elements of dementia syndrome,and observe the distribution of syndrome elements in patients with cognitive impairment,and the main syndrome elements were screened and their correlations among themselves and with cognitive impairment risk factors and scale scores were analyzed.Experimental part: To detect and analyze the effects of Jiedu Yizhi Formula on cognitive function,hippocampal pathology,intestinal flora,the expression of pyroptosis pathway-related proteins,Aβ1-42 and systemic inflammation in Alzheimer’s disease(AD)rats through Morris water maze,tissue sections,intestinal flora detection,Western-blot,immunohistochemistry,enzyme-linked immunosorbent assay methods,and explain the mechanism of Jiedu Yizhi Formula improving cognitive impairment in AD rats,and to provide an alternative therapy for AD.Methods: Clinical part: A cross-sectional questionnaire on the distribution of cognitive impairment syndrome elements in the elderly population in Changchun City was formulated,including general information,chronic disease history,self-rating depression scale,dementia syndrome element scale,simple mental state examination scale,Montreal Cognitive Assessment Scale,Grip Strength Measurement Scale,Activities of Daily Living Scale,and related examination records.The researchers were trained on related scale scoring and information evaluation of the four diagnosis of Chinese medicine.The information of patients aged 60-75 years old in some departments of the two affiliated hospitals of Changchun University of Chinese Medicine was collected and grouped for statistics.2.Morris water maze was used to monitor the behavior of rats.The water maze test was started after the gavage treatment.Through positioning navigation experiment and space exploration experiment,the effect of Jiedu-Yizhi Formula on the learning and memory function of AD rats was analyzed.Experimental part: 1.The AD model was established by injecting Aβ25-35 into the hippocampus of SD rats.After modeling,they were divided into model group,donepezil hydrochloride group,and Jiedu Yizhi Formula high,medium and low dose groups,and the same batch of rats were taken.as a blank group.Afterwards,the rats were given normal saline,Donepezil hydrochloride suspension,high,medium and low doses of Jiedu Yizhi Formula and normal saline by gavage for 8 weeks.2.After gavage,Morris water maze was used to test the learning and memory function of AD rats,and evaluate the efficacy of Jiedu Yizhi Formula.3.HE staining was performed on the hippocampus and liver tissues of rats to analyze the effect of Jiedu Yizhi Formula on the pathological tissue of the hippocampus of AD rats,and to observe whether Jiedu Yizhi Formula had liver toxicity.4.16 S intestinal flora sequencing to evaluate the effect of Jiedu Yizhi Formula on the structure and diversity of intestinal flora in AD rats.5.Western-blot method and IHC method were used to detect the expression levels of related proteins and Aβ deposition in the canonical NLRP3/Caspase-1/GSDMD pathway and non-canonical LPS/Capase-11/GSDMD pathway in the hippocampus of AD rats.6.Enzyme-linked immunosorbent assay was used to measure the expression levels of Aβ1-42,IL-1β and IL-18 in hippocampus,cerebral cortex,colon and serum of AD rats.Results: Clinical part: The main syndrome elements of patients with cognitive impairment(including mild cognitive impairment and dementia)are kidney deficiency,spleen deficiency,marrow reduction,phlegm turbidity,and blood stasis.Among the main syndrome elements,kidney deficiency is closely related to hypertension,and phlegm turbidity is closely related to diabetes and stroke.The five main syndrome elements are interconnected,and it was significantly negatively correlated with MMSE and Mo CA scores.The increase of kidney deficiency,marrow hypoplasia,phlegm turbidity,and blood stasis syndrome element scores had a causal relationship with the decrease of MMSE and Mo CA scores.The research results support the theory that "marrow deficiency and toxin damage" is the main pathogenesis of cognitive impairment patients,and "" Spleen and Kidney Deficiency,Phlegm Obstructing the Brain Syndrome "" is the main syndrome of cognitive impairment patients.Experimental part: 1 Morris water maze: compared with the blank group,the escape latency of AD model group was significantly prolonged,and the number of crossing the platform and the swimming time and distance in the platform quadrant were reduced.The high,medium and low doses of Jiedu Yizhi formula shortened the escape latency of AD rats and increased the number of crossing the platform and the swimming time and distance in the platform quadrant(P < 0.05).2.HE staining of hippocampal tissue and liver: neurons in hippocampal CA1 area of rats in the blank group are closely arranged,with clear and countable layers,normal and complete structure,abundant cytoplasm,round nucleus and normal staining.In the model group,the neuronal cells were disorderly arranged,the number of layers was unclear and the number decreased significantly,the morphology changed,the boundary between cytoplasm and nucleus was unclear,the nucleus pyknosis and staining deepened.Jiedu Yizhi formula can significantly restore the number and morphological structure of hippocampal CA1 cells,especially in the low-dose group.Liver slices showed that Jiedu Yizhi formula did not cause significant changes in the liver structure of rats.3.Sequencing results of intestinal flora: compared with the blank group,the diversity of intestinal flora of AD rats has not changed significantly,but the species structure has changed significantly.In the model group,the abundance of pathogenic bacteria has increased and the abundance of probiotics has decreased in the six biological classification levels of phylum,class,order,family,genus and species,such as the abundance of anaerobiospirillum has increased and the abundance of lactobacillus has decreased,while in the high,medium and low dose groups of Jiedu Yizhi formula,In particular,the low-dose group inhibited pathogenic bacteria and increased the abundance of probiotics,such as lactobacillus-johnsonii and lactobacillus-reuteri.4.Western blot results:the expression levels of NLRP3,caspase-1 and capase-11 precursors,shears and gsdmd in the hippocampus of AD rats were significantly higher than those in the blank group.Jiedu Yizhi formula decreased the expression of pyroptosis-related proteins,especially in the low-dose group(P < 0.05).5.Immunohistochemical results: Compared with the model group,Jiedu Yizhi formula reduced the Aβ deposition level,NLRP3 and GSDMD positive cells in the hippocampus of AD rats,and the low dose group was the most significant(p<0.05).6.ELISA results: Compared with the model group,Jiedu Yizhi formula reduced the expression levels of Aβ1-42,IL-1β and IL-18 in the hippocampus,cerebral cortex,colon and serum of AD rats.Conclusion:1.The main syndrome elements of mild cognitive impairment and dementia patients in Changchun City are kidney deficiency,spleen deficiency,marrow reduction,phlegm turbidity and blood stasis."Medullary deficiency and toxic damage" is universal as a key pathogenesis of patients with cognitive impairment.2.Jiedu Yizhi formula can increase the hippocampal neurons of AD rats,repair the pathological structure of the hippocampus,improve the intestinal flora disorder,reduce the deposition of beta amyloid,and down-regulate the expression of pyroptosis-related proteins in the hippocampus.It inhibits neuroinflammation and systemic inflammatory response,and improves cognitive dysfunction in AD rats.