| Objective:Aging is a degenerative change in the body with age.Non-healthy aging will increase the prevalence of neurodegenerative diseases associated with aging in the elderly,causing cognitive dysfunction.As the global aging population intensifies,the risk of senile diseases among elderly people also increases.Based on this,researching effective drugs to improve aging,prevent aging diseases caused by aging,and improve the quality of life of the elderly is the main work of aging research.Traditional Chinese Medicine believes that the increasing deficiency of kidney is the root of aging,so replenishing kidney and filling essence is an important method to delay aging.Achyranthes bidentata is sweet and slightly warm,it enters the liver and kidney channels,it has the effect of nourishing the liver and kidneys,strengthening muscles and bones.It is widely used in ancient and modern anti-aging prescriptions.Therefore,in this study,D-galactose was used to establish an in vivo and in vitro aging model of mice and neural stem cells,that comprehensively evaluate the efficacy of Achyranthes bidentata to interfere with aging,and use UPLC-MS/MS to conduct a component study on the efficacy components,that exploring the possible components of Achyranthes bidentata to delay aging.Method:First,the D-galactose-induced mice aging model was injected with a large dose for 10 consecutive weeks,and low,medium and high doses of Achyranthes bidentata extract were given at the same time as the model.The daily state,weight and organ index of the mice during the experiment were observed.The water maze experiment was used to evaluate the learning and memory abilities of the mice,and the open field experiment was used to analyze the autonomous activities and exploration behaviors of the mice.Neuronal pathological changes were analyzed with followed by the HE staining method for the hippocampus of the mice.Secondly,Achyranthes bidentata is separated by alcohol extraction and water precipitation,extraction,macroporous resin and gel column method.It used neural stem cells by primary culture as a carrier,using CCK-8 detection to track the proliferative activity,that applied to aging neural stem cells to evaluate drug efficacy.Finally,the traced active components were analyzed to explore the possible components of Achyranthes bidentata to delay aging through UPLC-MS/MS.Results:The mice in the model group showed signs of reduced weight,drowsiness,sluggish movement,dull hair,and reduced food intake.Compared with the model group,the daily status of the positive drug and Achyranthes bidentata dose groups were improved.The organ index of the brain and kidney of the model group decreased(p<0.05),and the mid-dose group of Achyranthes bidentata significantly increased the organ index of the brain and kidney(p<0.05).In the open field experiment,each dose group of Achyranthes bidentata increase the activity time of the central area and the number of standing hind legs(p<0.05).In the water maze experiment.the mid-and high-dose groups of Achyranthes bidentata can significantly shorten the escape latency of mice in directional navigation experiments(p<0.05).Each dose group of Achyranthes bidentata can improve the residence time and crossing of the original platform quadrant of the space search experiment in mice(p<0.05).In the reverse platform experiment of different doses of Achyranthes bidentata,the escape latency of finding new platforms was significantly shortened(p<0.05).Histopathological analysis of the hippocampus showed that the neurons of the model group were loosely arranged,the density and number of neurons were reduced,the number of cell layers was reduced,the gap was increased,the cytoplasm was darker in color,the nucleus of the neurons contracted,and degeneration and necrosis;The hippocampal neurons in each dose group of Achyranthes bidentata improved,the damage was alleviated,the density and number of neurons increased,the gap decreased.neurons were apoptotic.Neural stem cells were successfully cultured by primary culture,and its aging model was established through D-galactose.In activity tracking,Achyranthes bidentata can promote the proliferation of neural stem cells(p<0.05),and its n-butanol fraction is separated by Sephadex LH-20,the separation fraction(nB-Fr.a.2)promoted the proliferation of neural stem cells better(p<0.05).The nB-Fr.?.2 also improved the survival rate of senescent neural stem cells(p<0.05).The nB-Fr.?.2 was analyzed by UPLC-MS/MS and it was mainly composed of triterpenoid saponins.Conclusion:For the first time,the experiment used the "kidney-brain theory" to research anti-aging effects of Achyranthes bidentata.The results show that Achyranthes bidentata can improve the learning and memory abilities,autonomous activities and exploration abilities of aging mice,it can reduce neuronal apoptosis in hippocampus and improve neuronal damage.And it can promote the proliferation of normal and aging neural stem cells.It verified the anti-aging prescription theory of "tonifying the kidney,filling the essence and regenerating the brain marrow" in Traditional Chinese Medicine.For the first time,it provided experimental basis for Achyranthes bidentata in the anti-aging research by the "kidney-brain theory".At the same time,the activity tracking method was used to trace the active components of Achyranthes bidentata to anti-aging and neuroprotective effects on neural stem cells.For the first time,it has grasped the material basis of Achyranthes bidentata to promote the proliferation of neural stem cells,which provides a certain theoretical basis for guiding clinical precise medication. |
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