| ObjectivePolygonati Rhizoma is first published in the "Mingyi bielu",raw Polygonati Rhizoma numbs the tongue and stings the throat,while prepared Polygonati Rhizoma can remove the numbness of the tongue to avoid irritation,at the same time enhance its functions of invigorating the spleen,moistening the lungs and tonifying the kidneys,and wine preparation can also help its medicinal potential to better exert its tonic effect.In this study,the anti-aging activity of the Polygonati Rhizoma decoction before and after processing was investigated by using Caenorhabditis elegans(C.elegans)as a biological model to enrich the scientific connotation of the potency of the processing of Rhizoma Polygonati.By extracting and isolating the polysaccharide,the main active ingredient in Polygonati Rhizoma,the monosaccharide composition of polysaccharides before and after processing of Polygonati Rhizoma was studied,and the anti-aging activity and mechanism of Polysaccharide in raw Polygonati Rhizoma(RPRP)and Polysaccharide in prepared Polygonati Rhizoma(PPRP)were investigated to lay the foundation for further in-depth study on the aging mechanism of Polygonati Rhizoma polysaccharides.Methods1 Study on the anti-aging activity of the Polygonati Rhizoma decoction and Polygonati Rhizoma of processed yellow rice wine decoctionThe raw Polygonati Rhizoma and Polygonati Rhizoma of processed yellow rice wine were prepared according to the method of "Pharmacopeia of the People’s Republic of China"(2020 edition).The raw Polygonati Rhizoma and prepared Polygonati Rhizoma were soaked in an appropriate amount of water,then decocted in 8 times,6 times and 4 times the amount of water,extracted and filtered while hot,combined with the filtrate,concentrated under reduced pressure and freeze-dried.Wild type N2 nematodes were used for life span,lipofuscin,pharyngeal pump,locomotion and growth and reproduction experiments to investigate the aging retarding activity of the Polygonati Rhizoma decoction and Polygonati Rhizoma of processed yellow rice wine decoction.2 Analysis of the chemical composition of the RPRP and PPRPThe powder of raw Polygonati Rhizoma and prepared Polygonati Rhizoma were extracted by adding 10 times the amount of 95%ethanol and then filtered,the filtrate was concentrated to a certain volume under reduced pressure,added 4 times the amount of anhydrous ethanol at 4℃ for 12 h for alcohol precipitation,and then filtered.The filtrate is the crude RPRP and the crude PPRP.The crude RPRP and the crude PPRP were dissolved by adding appropriate amount of water,adding Sevage reagent to remove protein,using NKA-9 macroporous resin to remove pigment,dialysis to remove small molecule impurities,concentrated under reduced pressure and freeze-dried to obtain RPRP and PPRP.The polysaccharide contents of RPRP and PPRP were determined by UV spectrophotometric method.The contents of mannose,rhamnose,glucose,galactose and arabinose substances in RPRP and PPRP were determined by PMP derivatization method,and the changes in the monosaccharide composition of RPRP and PPRP were compared.3 Study on the anti-aging activity and mechanism of RPRP and PPRPWild type N2 nematodes were used for life span,lipofuscin,pharyngeal pump,locomotion and growth and reproduction experiments to investigate the aging retarding activity of the RPRP and PPRP.The oxidative stress assay was conducted to simulate the peroxidation phenomenon generated by the increase of free radicals,and the effect of RPRP and PPRP on reactive oxygen species and antioxidant enzymes in nematodes was also measured to investigate the mechanism of Polygonati Rhizoma polysaccharides in delaying aging.The pathways of aging in nematodes were investigated by using transgenic nematode longevity assay and fluorescence expression assay as well as RT-PCR to determine the expression levels of daf-2,age-1,daf-16,sod-3,ctl-1 and skn-1 mRNA.Results1 Study on the anti-aging activity of the Polygonati Rhizoma decoction and Polygonati Rhizoma of processed yellow rice wine decoctionCompared with the blank group,the Polygonati Rhizoma decoction could significantly reduce the accumulation of lipofuscin in nematodes,and the Polygonati Rhizoma of processed yellow rice wine decoction could significantly prolong the life span of nematodes,reduce the accumulation of lipofuscin in nematodes,and increase the pharyngeal pump and movement frequency of nematodes,and both the Polygonati Rhizoma decoction and Polygonati Rhizoma of processed yellow rice wine decoction had no adverse effects on the growth and reproduction of nematodes.2 Analysis of the chemical composition of the RPRP and PPRPThe content of RPRP was 87.86%and that of PPRP was 84.06%.The content ratio of five monosaccharides(mannose,rhamnose-rhamnose-glucose,galactose and arabinose)changed after the processing,and the content of mannose,rhamnose,galactose and arabinose increased while the content of glucose decreased according to the total amount of five monosaccharides.3 Study on the anti-aging activity and mechanism of RPRP and PPRPCompared with the control group,RPRP could significantly reduce the accumulation of lipofuscin in nematodes,and PPRP could significantly prolong the life span of nematodes,reduce the accumulation of lipofuscin in nematodes,and increase the pharyngeal pump and movement frequency of nematodes,and both RPRP and PPRP had no adverse effects on the growth and reproduction of nematodes.The experimental results of the mechanism study showed that PPRP significantly increased the survival rate and average lifespan of nematodes in the oxidized environment of juglone,which prolonged the maximum lifespan of nematodes.Meanwhile PPRP could significantly increase the activity of SOD and CAT in nematodes,and both RPRP and PPRP could reduce the level of ROS in nematodes.In addition,Polygonati Rhizoma polysaccharides significantly reduced the expression of daf-2 and increased the expression of daf-16 and sod-3.ConclusionThe evaluation of the activity based on C.elegans proved that both the Polygonati Rhizoma of processed yellow rice wine decoction,which simulated clinical application,and the medicinal part,PPRP,were superior to raw Polygonati Rhizoma in terms of longevity,lipofuscin,pharyngeal pump frequency and movement frequency,reflecting better anti-aging effects,providing a basis for the enhanced tonic effect of Polygonati Rhizoma after processing.The content of monosaccharides such as galactose in the polysaccharide varied considerably after processing of Polygonati Rhizoma.It is hypothesized that the enhanced anti-aging effect of Polygonati Rhizoma after processing is related to the change in the monosaccharide content of the polysaccharide.The polysaccharides of Polygonati Rhizoma can delay the aging of nematodes by increasing the content of antioxidant enzymes in nematodes,reducing the level of ROS and improving the resistance to oxidative stress.The mechanism of action was related to daf-2,sod-3 and daf-16 of the IIS pathway.This study provides a basis for the interpretation of the scientific connotation of the efficacy of the processing of Polygonati Rhizoma from the perspective of anti-aging,and the obvious anti-aging effect of Polygonati Rhizoma polysaccharides,especially PPRP,provides a basis for a more scientific and rational clinical use of Polygonati Rhizoma and related product development. |
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