Study On The Preparation Process Optimization Of Abscisic Acid And Its Anti-hepatic Fibrosis And Anti-aging Activities

Liver fibrosis is a chronic disease caused by long-term liver damage.If it is not controlled in time,it can lead to liver cirrhosis and even life-threatening.In addition,with the extension of human life span,the improvement of health awareness,and the pressure from all aspects of the increase in the aging population,delaying aging and improving the quality of life have become an important issue.Therefore,it is necessary to find a safe and effective drug with low side effects,and it is also a research direction of scientific researchers in recent years.Abscisic acid(ABA),a sesquiterpene component shared by plants and animals,is due to its good pharmacological activity in lowering blood sugar,inducing cancer cell apoptosis,anti-inflammatory,and improving atherosclerosis.Has received much attention.ABA has four mixed isomers.Compared with artificial synthesis,natural ABA has better activity,and natural abscisic acid can be obtained in a short time through fungal fermentation.Multi-bacterial fermentation has been widely studied and applied because it can overcome the adverse effects of single-bacterial fermentation that cannot be achieved or the fermentation conditions are relatively harsh,to obtain the target product quickly and efficiently.In this experiment,coix seed,black beans,red beans,mung beans,corn,and rice,which are used for medicine and food,were used as experimental materials,and the mixed fermentation process of Aspergillus nige and Botrytis cinerea was screened and optimized.And using the isolated and purified ABA as a medicine,the protective effects of TAA-induced liver fibrosis and D-galactose-induced aging mice were studied respectively.The specific results are as follows:1.Analyze the technical conditions of mixed bacteria fermentation through single factor experiment,and analyze the best fermentation production process of ABA in combination with response surface optimization method: fermentation time 5 days,Botrytis cinerea: Aspergillus niger 3:7 mixed culture,carbon source Glucose: sucrose 8:2,nutrient supplement corn meal 14g/L,the highest yield of fermented abscisic acid under these conditions.2.ABA intervention improved the survival rate of individuals during the experiment,increased liver index and body weight.H&E and Masson staining showed that ABA could significantly change the liver pathological changes caused by TAA.At the same time,ABA also reduces serum aspartate aminotransferase(AST),alanine aminotransferase(ALT)levels,liver malondialdehyde(MDA),and increases liver superoxide dismutase(SOD)and glutathione(GSH)levels.Level.Immunofluorescence staining showed increased expression of Caspase-3(Caspase-3),transforming growth factor β1(TGF-β1),and α-smooth muscle actin(α-SMA).In the immunoblotting results,the inflammatory protein NF-kappa B p65(p65)and interleukin 1β(IL-1β)levels caused by the TAA model group increased,and the inhibitory kappa protein(Ik B-α)decreased.ABA effectively inhibited the TAA-induced Inflammatory protein expression.ABA also regulates the expression of the apoptotic protein B lymphocyte tumor-2(Bcl-2),indicating that ABA inhibits TAA-induced liver fibrosis through anti-inflammatory and anti-apoptotic.3.ABA intervention for 5 weeks continuously reversed the changes in the levels of superoxide dismutase(SOD),malondialdehyde(MDA)and catalase(CAT)in mice serum caused by D-Gal,suggesting the level of oxidative stress in the body Be suppressed.The content of ACh E in brain tissue and the results of H&E staining indicated that ABA inhibited D-Gal-induced brain damage.At the same time,Western blot results showed that ABA can inhibit the increase of caspase-3(Caspase-3)and p53,regulate the protein levels of phosphoinositide-3-kinase(PI3K),protein kinase B(Akt),and Bcl-2 The expression of related X protein(Bax),B lymphoma-2(Bcl-2),sirtuin 1(SIRT1),and adenylate-activated protein kinase(AMPK),but it had no significant effect on the intestinal flora of mice influences.In summary,it is indicated that ABA can mediate inflammation and mitochondrial apoptosis and inhibit TAA-induced liver fibrosis.Inhibit D-Gal-induced subacute senescence by reducing apoptosis and DNA damage.The above experimental results provide references for the application of ABA in the future.