Preparation Of Shen-Rong Peptide Dropping Pills And The Effect Of Anti-Aging And Nootropic Effect Of Aging Mice

Objective:Aging is an irreversible natural law in the process of biological growth and development.With the increasing severity of population aging,the demand for delaying and preventing aging is increasing.In this study,the total saponins of ginseng stem and leaf were combined with deer antler polypeptide to study the best preparation process,quality standard and in vitro absorption of SRDP,and to explore the anti-aging and intelligence effect of SRDP on aging mice,in order to provide theoretical basis for the application and development of the TCM product.Methods:1.Preparation process of SRDPThe effects of matrix,condensing agent,PEG4000:PEG6000,drug:matrix,dropping distance and dropping speed on the formation of dropping pills were studied by single factor test,and the appearance quality of dropping pills,dissolution time limit and the coefficient of variation of pill weight were taken as the investigation indexes.Based on this,L₉（3⁴）orthogonal test was used to optimize the best preparation process of SRDP.Three batches of process verification were carried out.2.Quality standards SRDPAccording to the Chinese Pharmacopoeia（2020 edition）,the character,thin layer identification,weight difference,dissolution time of SRDP were examined,and the contents of ginsenoside Rg₁,Re,Rd and total protein in the dropping pills were determined to establish the quality standard of SRDP.3.Absorption of SRDP in vitroDetect the content of ginsenosides in artificial artificially pepsin and artificial intestinal fluids by HPLC,and analyze the protein of deer antler polypeptide by SDS-PAGE electrophoresis to investigate the stability of SRDP;Establish an everted gut sac,and investigate the absorption promoting effect of the matrix on ginsenosides by detecting the content of ginsenosides Rg₁,Re,Rb₁,Rc,Rb₂,Rd in the everted intestinal sac model and analyzing the absorption of deer antler peptides by SDS-PAGE electrophoresis.4.Anti-fatigue effect of SRDPThe results of pharmacodynamic experiment showed that the weight of mice increased in the same trend during the experiment.The results of weight-bearing swimming and pole climbing tests showed that SRDP could improve fatigue and poor coordination in aging mice caused by D-gal combined with aluminum trichloride.5.Anti-aging effect of SRDPMorris water maze experiment showed that SRDP could improve the aging signs of learning decline in D-gal combined with aluminum trichloride mice.The pathological results of brain tissue showed that SRDP could alleviate the problems of light-colored hippocampal cortex,disordered and sparse neuron arrangement,and shrinkage and reduction of Nishi bodies in the brain tissue of aging mice induced by D-gal combined with aluminum trichloride.The results of mouse brain biochemical kit showed that SRDP could improve antioxidant and metabolic capacity,increase T-SOD activity,and reduce MDA and LD contents in brain tissue.6.The effect of SRDP on anti-aging puzzle spectrum was analyzed based on grey correlation degreeGrey correlation analysis was used to investigate the correlation between ginsenoside Rg₁,Re,Rb₁,Rc,Rb₂,Rd and total protein content and T-SOD,MDA and LD in brain tissue of aging mice.Results:1.Process of preparation of SRDPThe results of single factor test showed that drug:substrate,drop distance,PEG4000:PEG6000 had great influence on the formation of dropping pills,and these three factors were selected for orthogonal test.The orthogonal test results showed that the factors affecting the process of SRDP were in the order of drop distance（cm）>drug:matrix>PEG4000:PEG6000,and the best process of SRDP was drug:The substrate ratio was 1:4,the drop distance was 15cm,PEG4000:PEG6000=1:2,the melting temperature of the substrate was 65℃,and the drop rate was 40 d/min.The results of verification experiment showed that the average dissolution time of the three batches of samples was 5.62 min,the coefficient of variation of pill weight was 3.074%,the comprehensive score was 4.209,and the RSD was 0.60%.2.Quality standards of SRDPGinseng antler peptide dripping pills are yellow brown,taste slightly bitter;The thin-layer identification results showed that the ginseng antler peptide dripper pills showed the same spots in the corresponding positions with ginsenoside Rg₁,Re and ginsenoside under the light and fluorescence,and the main spots in the corresponding positions with glycine,alanine,deer antler Control medicinal materials and deer antler polypeptide were the same color.The results of weight difference showed that the average pill weight of the three batches of samples（2021070701,2021070702,2021070703）was 0.0469 g,0.0449 g,0.0446 g,and the average dissolution time was 5.59 min,5.72 min,5.56 min,respectively.Meet the relevant requirements of the 2020 edition of the Chinese Pharmacopoeia;The content of ginsenoside Rg₁,Re and Rd were 0.91 mg/g,2.83 mg/g and 1.12 mg/g,respectively,with RSD of 1.34%,1.42%and 1.69%.The protein content was 11.68 mg/g,and the RSD was 2.67%,which met the standard.3.Absorption of SRDP in vitroThe stability experiment results of Shenrong Peptide Dropping Pills in artificial intestinal and gastric juices showed that the matrix of Shenrong Peptide Dropping Pills can protect the degradation of drugs in the gastric ring and delay the release of drugs in the gastrointestinal environment;The experimental results of the everted intestinal sac model suggest that under the action of the matrix,Shenrong Peptide Dropping Pills can promote the absorption of ginsenosides and deer antler peptides.4.Anti-fatigue effect of SRDPDuring the experiment,the weight increase trend of each group was consistent.The results of weight-bearing swimming and pole climbing tests showed that SRDP could improve fatigue and poor coordination in aging mice caused by D-gal combined with aluminum trichloride.5.Anti-aging effect of SRDPMorris water maze experiment showed that SRDP could improve the aging signs of learning decline in D-gal combined with aluminum trichloride mice.The pathological results of brain tissue showed that SRDP could alleviate the problems of light-colored hippocampal cortex,disordered and sparse neuron arrangement,and shrinkage and reduction of Nishi bodies in the brain tissue of aging mice induced by D-gal combined with aluminum trichloride.The results of mouse brain biochemical kit showed that SRDP could improve antioxidant and metabolic capacity,increase T-SOD activity,and reduce MDA and LD contents in brain tissue.6.The effect of SRDP on anti-aging puzzle spectrum was analyzed based on grey correlation degreeGrey correlation analysis indicated that the contribution of SRDP protein to T-SOD,MDA and LD was the largest,followed by ginsenoside Rd to T-SOD,MDA and LD.Conclusion:The preparation process of SRDP is stable,reasonable and feasible,and the established quality standard is stable and Controllable.It has anti-aging and intellectual effects on the aging mouse model induced by D-gal combined with aluminum trichloride.