Neurotrophic Effects Of Aqueous Extract From Juglans Regia L.

With the aggravation of the aging population,the elderly diseases are taken seriously.Neurodegenerative diseases seriously affect the normal life of patients,so should immediately solve such problems.Natural products have irreplaceable advantages,its wide range of sources,diverse structure and high security,is the new focus of drug development in recent years.Walnut(Juglans regia L)is a walnut branch,walnut is a perennial deciduous tree,scientific name walnut.Walnut in China has a long history of cultivation,North China,northwest,southwest,central China,southern China and east China,southern Xinjiang and other 22 provinces are large area of cultivation.Walnut is rich in oil,protein and a variety of trace elements.In recent years,more studies are oleic acid,linoleic acid and linolenic acid.A large number of literature,experimental results show that unsaturated fatty acids can lower cholesterol,delay nerve aging and promote nerve regeneration.Whether walnut kernel aqueous extract has neurotrophic and neuroprotective activity is the focus of this paper.The walnut peeled,beaten,degreasing,adding water,ultrasonic extraction,steaming,dissolved in water made of 2mg/mL of the mother liquor.The neurotrophic effects of the extract were evaluated by using neuroblastoma cell line(SH-SY5Y)cell line in vitro.And then detected the neurofilament protein(NF160)gene expression and protein expression.The main conclusions are as follows:(1)Walnut aqueous extract stability experiments show that the water extract in the high temperature will be degraded.The aqueous solution was prepared by HPLC at 4 ?,25 and 37 ? ?for 1 day,2 days and 3 days respectively.Then,the chromatographic fingerprints were evaluated by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine.The results showed that the water was degraded at 37 for 1 day,and there was such a phenomenon at ?25 ?,but it did not show obvious degradation at 4 ?.The pH of the aqueous extract was adjusted to 5.5,6.5,7.5 and 8.5,respectively.The results showed that there was no significant difference in the similarity evaluation of Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine.It is recommended that the water extract stored in the temperature below 4 ?,so that the sample can ensure that the degradation.(2)Unbleached walnut powder of the aqueous extract has obvious cytotoxicity,but peeled walnut powder aqueous extract no obvious cytotoxicity.The cells were treated with untreated aqueous walnut for 24 hours.MTT assay showed that the final concentration of aqueous extract was above 400 ?g/mL.The results of MTT test showed that there was no obvious cytotoxicity between the final concentration of aqueous extract from 0.64 ?g/mL to 10000 ?g/mL.(3)Walnut peeled skim extract has the activity of promoting synaptic elongation of SH-SY5 Y cells,and can up-regulate the expression of NF160 gene and NF160 protein expression.The cells were treated with peeled water for one week and observed by fluorescence microscopy.It was found that there was a significant increase in synapses at a final concentration of 5 mg/mL.The results of qPCR and Westren blot showed that the NF160 gene was up-regulated at the final concentration of 5 mg/mL,and the protein expression was also significantly improved.(4)Literature and labeling to determine the water containing the known components of cytidine,uridine,guanosine and adenosine.Cytotoxicity results showed that the final concentration of adenosine from 0.2 ?g/mL or more cytotoxicity,in addition to the other three nucleosides and no obvious cytotoxicity.The four nucleosides were mixed at the original ratio and the cytotoxicity test showed no significant toxicity.Cells were treated with four nucleosides for one week and then observed with a microscope.Cytidine,uridine and guanosine have the activity of promoting synaptic growth,respectively,at 20 ?g/mL,2 ?g/mL,0.8 ?g/mL have the best activity,but the length of synaptic growth is less than the aqueous extract.(5)HPLC showed that there was a large peak in the aqueous extract,the retention time was 37.5min.After collecting the peak,the cells were treated.The results of cytotoxicity showed that there was no obvious cytotoxicity between 1 ?g/mL and 1000 ?g/mL,And did not significantly promote synaptic growth.