Extraction And Purification Of Nervonic Acid And Effect On The Repair Of Myelin Sheath Injury In Mice With Encephalomyelitis

Nervonic acid is the core natural component of brain nerve tissue and nerve cells.It is a special effect substance discovered in the world to promote the repair and regeneration of damaged nerve tissue.The fatty acids of acer truncatum seed oil contain 5 to 6 percent nervonic acid,which is a new resource to realize nervonic acid mass production.In this paper,the extraction of acer truncatum seed oil,the isolation and purification of nervonic acids,and the effects on encephalomyelitis in mice were studied.1.Single factor and orthogonal experiments were used to optimize the extraction process of acer truncatum seed oil by subcritical fluid extraction(SFE)technology.The results showed that:the particle size of 60 mesh was crushed,extraction was carried out at 40℃ for 150 min and 4 times,the yield was 32.41%.Gas chromatography-mass spectrometry(GC-MS)analysis showed that acer truncatum seed oil contained 12 kinds of fatty acids,of which the content of nervonic acid was 5.15%.2.The process conditions for purifying nervonic acid by molecular distillation(MD),urea complexation(UC)and preparative high-performance liquid chromatography(Pre-HPLC)were optimized by single factor and response surface analysis.The results showed that:distillation temperature 145℃,vacuum degree 0.01 mbar,nervonic acid purity was 23.4%;on the basis of first-stage molecular distillation,six-stage molecular distillation was carried out,the nervonic acid purity was 40.2%;Under the condition of-14℃ and 13.5 h inclusion,the purity of nervonic acid was 15.8%;the molecular distillation method was combined with urea complexation method,and the purity of nervonic acid was 60.45%;the purified nervonic acid was purified by Pre-HPLC method,the final purity of nervonic acid was 95.8%.3.Experimental autoimmune encephalomyelitis(EAE)mice were selected as animal models.Low-,medium-,and high-dose of groups were gavaged with 197 mg/kg,394 mg/kg,and 788 mg/kg nervonic acid,and drug group with gavage 6 mg/kg acetic acid Prednisone,normal group and model group were given the same amount of normal saline.The results showed that:nervonic acid had effect on improving the memory and learning ability of mice.At the same time,nervonic acid can reduce inflammation,improve the symptoms of myelination,increase the number of positive cells of NQO1,Nrf2 and HO-1,increase SOD and CAT Activity,reduce ROS and TBAR levels,increase the expression level of anti-inflammatory factors IL-4/IL-10,and reduce the expression level of pro-inflammatory factors IFN-y/TNF-α.Nervonic acid changed the balance between pro-inflammatory and anti-inflammatory cytokines in the body and formed an anti-inflammatory microenvironment in the central nervous system.