| Aging is an irresistible natural process in life activities.In the research of anti-aging natural medicines,it has been found that Hippophae rhamnoides Polysaccharides and ginsenosides have good anti-aging effects.However,most studies have focused on the anti-aging effects of using H.rhamnoides Polysaccharides or ginsenosides alone.There is little research on the anti-aging activity of the combination of them.In this study,response surface methodology was used to optimize the compound enzymatic method(cellulase: pectinase=1:1)to extract H.rhamnoides Polysaccharides.And then the H.rhamnoides Polysaccharides was precipitated and grouped by using absolute ethanol.While the ginsenosides was quickly extracted by ultrasonic extraction method,purified and grouped by using D101 macroporous resin.Secondly,the optimal antioxidant polysaccharides group and ginsenoside group were screened by the in vitro antioxidant assays.Furthermore,they were matched according to the ratio of 1:1,and then screened the safe dosage of the proportioning compound.Finally,Caenorhabditis elegans was used as the natural aging model to study the anti-aging effects of the proportioning compound.And the Illumina Miseq sequencing technology was used to investigate the gut microbes structure of C.elegans after administration.The main findings are as follows:1.The optimized preparation process was obtained through the response surface method.And the optimal conditions were determined as follows: the liquid-to-material ratio is 20:1,the enzyme concentration is 2.4%,and the enzymatic hydrolysis time is 150 min.Under these conditions,the extraction rate of H.rhamnoides Polysaccharides was 3.89%.The influence of the three factors on the yield of polysaccharides from large to small is compound enzyme concentration>enzymolysis time>liquid-to-solid ratio.In addition,the interaction between liquid-to-solids ratio and enzymatic hydrolysis time has the greatest impact on the yield.2.Among the 5 polysaccharide groups and 2 ginsenoside groups,the 80%ethanol fractional precipitation group and the 40% ethanol separation component have the best antioxidant activity.Then they were combined to form the proportioning compound.And three gradient concentrations(0.9 mg/m L,1.8 mg/m L,and 2.7mg/m L)were set for the compound to investigate the influence on C.elegans aging model.Compared with the control group,at a dose of 2.7 mg/m L,the cumulative level of lipofuscin,the level of ROS and the lipid content in C.elegans decreased by72.99%,64.5% and 71.02%,respectively.The average lifespan of C.elegans was extended by 36.1%,and the SOD and CAT enzyme activities in the body were increased by 1.88 and 2.39 times,respectively.3.Gut microbiota analysis revealed that the proportioning compound has the effect of improving microbiota composition in C.elegans.At the genus level,the abundance of the probiotic Lactobacillus and the harmful bacterium Pseudoxanthomonas increased and reduced,respectively.The results of this study showed that H.rhamnoides Polysaccharides and ginsenosides have a synergistic anti-aging effect on C.elegans model.Moreover,the effects of using them in combination can effectively improve the gut microbes structure of C.elegans.And increased the abundance of probiotics and reduced the abundance of harmful bacteria.The development of this research provides a certain theoretical basis for the combined use of H.rhamnoides Polysaccharides and ginsenosides in the future. |
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