Study On The Anti-Aging Effect And Components Of Gymnadenia Conopsea Based On Caenorhabditis Elegans And UPLC-QTOF-MS

Traditional Chinese medicine "shouzhangshen" is commonly used as a tonic and anti-aging medicine,but its efficacy lacks systematic evaluation and the effective substances urgently needs to be clarified.Research has shown that antioxidant activity is one of the effective ways to combat aging.In the present study,"shouzhangshen"extract was first separated by macroporous resin to obtain the corresponding fractions.Then,the total phenol and flavonoid contents of fractions were determined,and their antioxidant activities were evaluated,so as to screen out potential anti-aging active site.In addition,the anti-aging effects of the potential active site were comprehensively evaluated to identify the anti-aging active site of "shouzhangshen"by using Caenorhabditis elegans,a model organism.Further chemical composition analysis was conducted using UPLC-QTOF-MS,and the following research results were obtained.1.Four macroporous adsorption resins,D101,AB-8,ADS-17,and ADS-F8,were used to separate the extract of "shouzhangshen".The extract was eluted with water,15%,30%,50%,75%,and 100%ethanol,respectively,to obtain the corresponding elution sites.The total phenol and flavonoid contents of the elution sites were determined.The results showed that the 50%ethanol elution site(D50)of D101 macroporous resin had the highest total phenol and flavonoid contents.2.The antioxidant activities of various parts separated by D101 macroporous adsorption resin were evaluated by using DPPH and ABTS free radical scavenging assays.The antioxidant activity of each part from high to low was as follows:D50>30%ethanol elution site(D30)>75%ethanol elution site(D75)>15%ethanol elution site(D15).The water elution site(D0)and 100%ethanol elution site(D100)didn’t show antioxidant activities.3.Content determination and antioxidant activity results indicated that the D50 site might be the potential active site.Next,the anti-aging effect of D50 site was comprehensively evaluated by using the model organism C.elegans.The results showed that D50 could significantly prolong the lifespan of C.elegans in a dose-dependent manner,and the effect of 200 μg/mL of D50 was equivalent to that of the positive drug astaxanthin.In the reproductive capacity test,D50 increased the number of eggs laid by C.elegans,shortened the generation time,and improved the reproductive ability.In the athletic ability test,D50 increased the body bent frequency,head swinging frequency,and pharyngeal pump frequency and enhanced the exercise ability.In body parameter test,D50 increased the body length and width of the C.elegans.The above results indicated that D50 had a significant anti-aging effect.4.Chemical composition of D50 was characterized using UPLC-QTOF-MS.31 compounds were identified,including 8 phenolic compounds,7 succinate glycosides,5 alkaloids,3 flavonoids,2 terpenoids and steroids,1 stilbene,1 phenanthrene,and 4 other compounds.The present study provides modern scientific evidence for the traditional efficacy of "shouzhangshen" and provides a theoretical basis for its in-depth development.