Study On The Efficacy And Related Mode Of Action Of Traditional Chinese Medicine Formula Kang-Shuai-Lao-Pian

The aged population in the world is increasing,and researchers attach much importance to study aged related diseases.The goal of anti-aging research is not limited to prolonging the lifespan,but more importantly,to increase the healthy lifespan of individuals.In long-term medical practice,many traditional Chinese medicine prescriptions have been approved for the effect of delaying aging.Kang-Shuai-Lao-Pian(KSLP)is originated from the secret recipe of Yishou-Yongzhen Ointment of Ming Dynasty,which is composed of Rehmannia glutinosa,Panax ginseng,Asparagus cochinchinensis,Ophiopogon japonicus,Lycium chinense and Poria cocos.KSLP has the effect of nourishing qi and nourishing yin,soothing and reassuring.It is clinically used for mid-aged and elderly individuals who are weak due to qi and yin deficiency,fatigued,palpitations,shortness of breath,dizziness,hot flashes,night sweats,tinnitus forgetfulness,irritability and insomnia.However,its efficacy evaluation and mode of action need to be clarifiedBased on the pathological and metabolic characteristics of aging,this study evaluated the efficacies of KSLP on mid-aged mice,elderly mice and high fat diet(HFD)induced obese mice.In order to reveal the mode of action of KSLP,we applied 16S rRNA gene sequencing and non-target fecal metabolomics techniques to analyze the profiles of gut microbiota and fecal metabolites.Besides,RNA-sequencing(RNA-seq)based transcriptomic study was performed on adipose derived stem cells(ADSC).The main results are as follows1.KSLP significantly promoted brain dopamine production and increased the relative content of brown adipose tissue in mid-adult mice(8-months).KSLP increased gut microbiota diversity and improved the gut microbiota composition of mid-adult mice,showing that significantly increase dopamine production related-Clostridia and butyrate producing bacteria including Lachnospiraceae_NK4A136_group,Anaerotruncus,Butyricicoccus,Ruminococcaceae UCG-005,etc.The detection of gut butyrate showed that KSLP slightly increased butyrate contents.KSLP improve the neurotransmitter dopamine and fat metabolism disorders that occur in earlier aging stage,which may be achieved via regulating the gut microbiota and promoting butyrate production,and then affecting the gut-brain axis and gut-metabolism axis2.KSLP had no significant effect on the body weight and the distribution of white and brown adipose of aged mice(16-18 months),while improved the physiological status of aged skin,increased skin collagen and neurotransmitters contents of DA,NE,Ach,5-HT.In addition,KSLP significantly improved the differentiation function,especially the osteogenic differentiation abilities of aged adipose stem cells(ADSC)Analysis of RNA-sequencing data illustrated that the molecular mechanisms may be related with PI3K-Akt signaling pathway,mRNA surveillance,Cytokine-cytokine receptor,etc.Analysis of gut microbiome data demonstrated KSLP have no effect on the gut microbiota diversity,while increase the relevant abundance of Allobaculum and Romboutsia but decrease that of Faecalibacterium and Akkermansia in the gut of aged mice3.KSLP improved HFD induced obesity,glucose tolerance disorder,as well as gut dysbiosis in mice.The results analysis of gut microbiome showed that KSLP improved the gut microecological disorders of HFD mice.At phylum level,KSLP corrected the increased relative abundance of Firmicutes and Proteobacteria,and decreased abundance of Bacteroidetes caused by HFD.At family level,KSLP reversed the relative abundance of Ruminococcaceae and Desulfovibrionaceae.KSLP also reversed HFD induced significant changes in the abundance of certain genus including Intestinimonas,Oscillibacter,Christensenellaceae_R-7_group,Ruminococcaceae UCG-010 and Aliihoeflea.Pearson correlation analysis indicated that except for Ruminococcaceae UCG-010,other 4 genera had positive correlations with obesity.In addition,the metabolites in feces regulated by KSLP involve amino acid metabolism,peptide metabolism,and lipid metabolism,which are closely related with Intestinimonas,Oscillibacter,and Christensenellaceae_R-7_group.The above results hint that KSLP may through regulating obesity-related bacteria to affect the body metabolism,and then achieved the efficacy of preventing HFD induced obesity and metabolic disorders.