Development Of Rongshen Yizhi Tablets With Auxiliary Memory Improvement Function

Memory impairment refers to the phenomenon where individuals experience memory difficulties or loss of memory ability in their daily lives regarding past events.Memory impairment is caused by multiple factors,among which neurodegenerative diseases are an important factor leading to memory impairment.With the acceleration of the aging process in China,the proportion of the elderly people continues to increase.Among the population aged 60 and above,70%to 80%suffer from varying degrees of forgetfulness and dementia.It is estimated that by 2050,the number of global dementia patients worldwide will exceed 100 million,and the number of elderly dementia patients in China will reach 28.98 million.Currently,drugs such as donepezil,memantine,and nimodipine used to treat Alzheimer’s disease（AD）can only alleviate early symptoms of patients,while also having significant side effects and high long-term medication costs.Traditional Chinese medicine（TCM）,however,has the characteristics of multiple components,multiple targets,and multiple pathways,with most TCM formulas being relatively inexpensive.Therefore,the development of TCM as health food for improving memory function has promising prospects.The Rongshen Yizhi Decoction is a clinical experience formula from Changchun University of Traditional Chinese Medicine,mainly used to improve learning and memory function and prevent Alzheimer’s disease.This formula consists of ginseng,Poria cocos,Polygala tenuifolia,deer antler,and deer heart.Ginseng refers to the dried roots and rhizomes of Panax ginseng C.A.Mey.,a plant of the Araliaceae family.Its main active ingredient is ginsenosides,which have anti-inflammatory,anti-aging,neuroprotective,and memory-enhancing effects.Poria is the dried sclerotium of the fungus Poria cocos（Schw.）Wolf,which belongs to the Polyporaceae family.Its main active ingredients include triterpenes,fatty acids,and Poria polysaccharides,which have anti-inflammatory,antioxidant,and memory-improving effects.Polygala is the dried root of Polygala tenuifolia Willd.or Polygala sibirica L.,both plants of the Polygalaceae family.It contains various effective ingredients such as Polygalasaponins and flavonoids,which have the ability to improve learning and memory,anti-aging and neuroprotection effects.The heart of the sika deer（Cervus nippon Temminck）or red deer（Cervus elaphus Linnacus）is called Lixin,a commonly used medicinal material in Jilin Province.It contains rich active ingredients such as proteins and essential amino acids that are beneficial to the human body.Traditional Chinese medicine believes that Lixin has the effects of nourishing qi and blood,and calming the mind.Velvet antler is the immature antler of the male sika deer（Cervus nippon Temminck）or red deer（Cervus elaphus Linnaeus）covered with dense fur.It contains multiple effective ingredients such as peptides,inorganic elements,and fat-soluble components,which have antioxidant,immune-enhancing,and memory-enhancing effects.This study aims to improve upon the basis of Rongshen Yizhi Decoction and develop it into Rongshen Yizhi Tablets,which have the advantages of stable quality,convenient portability,and easy large-scale production.However,the quality standards,efficacy ingredients,and mechanism of action for improving memory are not clear.This study investigates Rongshen Yizhi Tablet from these aspects,and the results are as follows.1.The optimal preparation process for the Rongshen Yizhi tablet is as follows:clean Poria and Polygalae radix,add 10 times the amount of water,boil three times,each time for 2 hours,filter,and concentrate the filtrate into paste.Use vacuum drying to obtain paste powder,sieve the paste powder through a 100-mesh sieve,mix it evenly with ginseng,deer heart,and deer antler micro-powder that have been crushed and sifted through a 100-mesh sieve.Add 30%of maltodextrin,1.4%of citric acid,and 40%of xylitol according to the prescription,mix well,and use 95%ethanol as a wetting agent.Wet granulation through a 20-mesh sieve,dry the obtained particles under vacuum at 55℃until the water content is around 4-8%through a 14-mesh sieve.Finally,mix the obtained particles with 1%of magnesium stearate according to the prescription and press them into tablets.The hardness of the tablets is between 57-65N,and the differences in tablet weight,microbial limit,and pollutant limit are within the specified range.Meanwhile,the content of ginsenosides Rg₁,Re,Rb₁,and polygalasaponins in the Rongshen Yizhi tablet was determined by HPLC,and the quality standards for the Rongshen chewable tablet were formulated in accordance with the national standard for health food safety（GB16740-2014）.2.A total of 61 effective components of Rongshen Yizhi tablets were screened using a combination of liquid chromatography-mass spectrometry and network pharmacology.These components were predicted to act on 63 key targets associated with memory impairment.It is speculated that Rongshen Yizhi tablets improve memory impairment by modulating cell apoptosis,neuroprotection,and immune regulation through signaling pathways such as PI3K-Akt,c AMP,and neuroactive ligand-receptor interactions.This provides a basis for further investigation into the mechanism of action.3.Safety evaluation of the Rongshen Yizhi tablets.The results of the 14-day acute oral toxicity test showed that the oral median lethal dose(LD₅₀)of the Rongshen Yizhi tablets in KM mice of both sexes was greater than 10 g/kg BW,and according to the dose classification standard of GB 15193.3-2014"National Food Safety Standard Acute Oral Toxicity Test,"the sample belongs to the practically non-toxic level.The results of the 28-day oral toxicity test showed that the Rongshen Yizhi tablets had no significant damage to important organs such as the heart,liver,spleen,lungs,kidneys,and brain of female and male mice,indicating that the Rongshen Yizhi tablets comply with the national safety standards for health food and can be promoted.4.Evaluation of the efficacy of the Rongshen Yizhi tablets.The behavioral experiments showed that the Rongshen Yizhi tablets could significantly improve scopolamine-induced learning and memory impairment.The 1 g/kg,2 g/kg,and 4g/kg doses of the cognitive-enhancing chewable tablets could significantly increase the ACh and Ch AT content in the brain tissue of scopolamine-induced mice,while significantly decreasing the level of ACh E.Treatment with the Rongshen Yizhi tablets inhibited the downregulation of the relative expression levels of p-PI3K/PI3K,p-AKT/AKT,Bcl-2/Bax,and upregulation of RXRA/GAPDH in scopolamine-induced mice,indicating that the Rongshen Yizhi tablets could significantly improve the learning and memory abilities of scopolamine-induced mice,possibly by regulating the PI3K/AKT pathway,increasing the expression of anti-apoptotic proteins,and enhancing the acetylcholine content to improve learning and memory impairment in mice.5.The protective effect of serum containing Rongshen Yizhi tablets on LPS-induced BV2 cell damage was studied using a serum pharmacological combined with liquid chromatography-mass spectrometry experiment method.The results showed that the serum containing Rongshen Yizhi tablets inhibited the increase in pro-inflammatory cytokine levels and the decrease in anti-inflammatory factor content in LPS-induced BV2 cells.Meanwhile,it promoted the phosphorylation of PI3K and AKT proteins and the expression of anti-apoptotic proteins in LPS-induced BV2 cells,while down-regulating the expression level of pro-apoptotic proteins.This suggests that Rongshen Yizhi tablets can exert anti-inflammatory and anti-apoptotic effects by regulating the expression of related proteins,thus protecting neuroglial cells.