Effect Of Probiotics Combined With Donepezil Hydrochloride On Cognitive Function And Gut Microbiota Of Alzheimer’s Disease

Objective: There are few randomized controlled studies on probiotic intervention in the treatment of AD patients,and the results are inconsistent.Furthermore,All these studies were placebo-controlled studies to evaluate the effect of probiotics in the treatment of AD.It is not yet known whether the combination of probiotics and conventional treatment can improve the clinical efficacy of AD,and how probiotics affect the gut microbiota of AD patients needs to be further clarified.To evaluate the effect of probiotics combined with donepezil hydrochloride in the treatment of AD patients through randomized controlled trials,and to compare the differences in gut microbiota between healthy elderly and AD patients,as well as the changes in gut microbiota before and after treatment,so as to explore the possible mechanism and provide new clinical evidence for the treatment of AD.Methods: According to the inclusion and exclusion criteria,29 healthy elderly and 58 mild and moderate AD patients were recruited in the Second People’s Hospital of Guizhou Province.58 AD patients were randomly divided into donepezil hydrochloride(DH)group and donepezil hydrochloride combined probiotic(DP)group.Patients in the DH group were treated with donepezil hydrochloride tablets,while those in DP group were treated with donepezil hydrochloride tablets and mixed probiotics by 12 weeks.The general basic imformation as well as family history of the patients were collected through interviews and medical record.The mini-mental state examination(MMSE)and modified Hasegawa’s dementia scale(HDS-R)were used to assess the changes of cognitive function before and after the treatment.At the same time,the patients’ ability to live daily(ADL)was measure.Fecal samples of healthy subjects and AD patients were collected,and the gut microbiota was detected by 16 S r RNA sequencing technology.The Alpha diversity and Beta diversity of gut microbiota in different groups were analyzed by bioinformatic methods.Results: 1.Effects of probiotics combined with donepezil hydrochloride on cognitive function.The subjects were randomly divided into the DH group and DP group.There was no statistical difference in age,BMI,gender,educational level,complication,MMSE,ADL and HDS scores between the two groups.We compare the cognitive function scores of DP group and the DH group after 12 weeks of treatment and use analysis of covariance(ANCOVA)to adjust covaritate such as age,gender,and BMI at baseline.The MMSE(15.652±3.312>14.055±3.515)and HDS-R score(15.207±4.752>13.707±6.409)of DP group were higher than that of the DH group,and the difference were statistically significant(P<0.05).Although the ADL score(33.379±11.034<37.879±10.808)of the DP group was lower than that of the DH group,there was no statistical significance(P>0.05).The paired t test was used to compare the changes in cognitive function after 12 weeks of treatment.Compared with the baseline,the MMSE(15.652±3.312>15.069±3.515)and HDS-R scores(15.207 ± 4.752>14.138 ± 4.663)in DP group increased,and the differences were statistically significant(P<0.01).However,there was no significant change in ADL score(P>0.05).2.Analysis of the difference of gut microbiota between AD group and control group.The final OTU data for the AD and control groups consisted of 2732 OTUs,2140 for AD patients and 1453 for healthy elderly.Among them,there were 861 OTUs for healthy elderly and AD,and OTU abundance in AD patients increased significantly.At the family level,Veillonellaceae,Lactobacteriaceae,and Verrucomicrobiaceae were significantly increased(P<0.01)in AD group,while Enterobacteriaceae and Lachnospiraceae were significantly decreased(P<0.001)when compared with control group.Further analysis using LEf Se(LDA Effect Size)revealed Synergists Lactobacillus,Proteobacteria,Erysipelotrichi and methanobacteria were the key taxa of AD patients from class to genus level.There was no significant differences in Chao1,Shannon,Simpson’s index and Faith’s Phylogenetic Diversity between AD patients and healthy elderly.The Beta diversity was significant difference(P=0.003)between the AD groups and the healthy elderly group.Correlation analysis showed that MMSE had a significant positive correlation with Faecalibacterium and a significant negative correlation with Christensenella.ADL and Paraprevotella were positive correlation,as well as HDS was significantly negatively correlated with Porphyromonas and Finegoldia.3.Effects of DH and DP on gut microbiota.There were significant changes in gut microbiota in phylum,class,order and family level between the DH group and DP group.Compared with the healthy elderly group,Lactobacillus,Enterococcus,Turicibacterales,Veillonellaceae and Christensenellaceae were significantly increased before donepezil hydrochloride treatment(P<0.05)and the Ruminococcaceae decreased(P<0.05).Fusobacteriaceae,Desulfovibrionaceae and Peptostreptococcaceae were significantly increased(P<0.05),while Veillonellaceae,Lactobacillus and Enterococcaceae were decreased significantly(P<0.05),as well as Ruminococcaceae was increased after treatment.Moreover,Lactobacillus,Veillonellaceae and Ruminococcaceae were no significant difference compared with control group.Lactobacillus enterococci was increased,as well as Turicibacterales and Lactobacillus luciferotidae were decreased after treatment with probiotics combined donepezil hydrochloride(P<0.05).Althouth Ruminococcaceae was increased and there was no significant difference compared with healthy elderly after the combined treatment.Lactobacillus and Eucobacteriaceae were also decreased compared with healthy elderly.Although the diversity of Chao1,Shannon,and Faith’s phylogenetic diversity compared with healthy controls were decreased after 12 weeks,it was not statistically significant.The heterogeneity was close to that of the healthy elderly after treatment,although there was no statistical difference among all groups.In addition,whether the DH group or the DP group,or control group(P<0.01),there was no significant change in Beta diversity before and after treatment.4.Conclusion: 1.The combination treatment of probiotics and donepezil hydrochloride can improve the cognitive function of patients with mild and moderate AD.2.There is a certain difference in the composition of gut microbiota between AD patients and healthy elderly.Both donepezil hydrochloride and the combination of probiotics and donepezil hydrochloride can promote the recovery of gut microbiota proportion,but there is no significant change in Alpha diversity and Beta diversity.