Hexacosanol Preparation From Barley Seedlings And Its Effect On Fatty Liver Injury

The barley seedlings showed good efficacy in anti-inflammatory,hypoglycemic and lipid-lowering activities.As an effective active ingredient detected in barley seedlings,hexacosanol is very likely to participate in the regulation of some physiological functions of barley seedlings.In this paper,the barley seedlings from Diqing,Yunnan Province were used as research objects,and the extraction process was used to prepare BS-Hex(Hexacosanol extracted from Barley Seedlings).Based on this,we explored the anti-oxidation ability of BS-Hex.Further,we constructed an in vitro model to study the improvement of cell lipid peroxidation injury induced by palmitic acid(PA),combined with in vivo animal experiments to evaluate the protective effect of BS-Hex on acute alcoholic liver injury in mice.It aims to provide basic theoretical support for the application and development of green barley,plateau special crops.The main research results are as follows:(1)Extraction of BS-Hex with sodium hydroxide ethanol solution as extractant,the process optimization results are as follows:extraction ratio of material to liquid(w/v)1:10,extraction time of 3.0 h,extraction temperature of 85 ?,oxidation The sodium ethanol concentration was 8%,and 700±20 mg BS-Hex could be extracted per 100 g of barley seedlings powder(dry weight,DW)under the optimal process conditions.By comparing the results of peroxide value(POV)measurement,more than 16 mg BS-Hex has good anti-oxidation ability,which is equivalent to the effect of 8 mg of common food antioxidant t-butyl hydroquinone(TBHQ).(2)The lipid peroxidation injury model of HepG2 cells induced by palmitic acid(PA)was successfully constructed in this study.The induction time was 12 h,and the PA concentration was 0.10 mmol/L,was the best modeling condition.At this time,the cells were obviously injured.The morphogenesis becomes smaller and wrinkled,the edge is not clear or even rounded off;the intracellular reactive oxygen species(ROS)fluorescence intensity is significantly enhanced;the intracellular lipid droplets are accumulated;the contents of triglyceride(TG)and malondialdehyde(MDA)increased to 1.21 times and 4.29 times,respectively,of normal cells.(3)It was found that BS-Hex can improve lipid peroxidation(LPO)injury in HepG2 cells.The results showed that the morphology of LPO-injured cells was significantly improved after adding BS-Hex.Compared with the model group,BS-Hex(20,40,60 ?g/mL):1.reduced intracellular ROS levels by 34.16%and 55.54%.60.89%;2.increased the activity of superoxide dismutase(SOD)by 82.80%,73.45%,74.85%;3.increased the activity of glutathione peroxidase(GSH-px)by 27.15%,44.59%,44.37%;4.decreased the content of alanine aminotransferase(ALT)in culture supernatant by 66.49%,59.43%and 74.86%;5.decreased the content of aspartate aminotransferase(AST)in culture supernatant by 60.65%,69.74%and 55.72%;6.decreased the level of tumor necrosis factor(TNF-a)by 22.99%,49.87%,and 57.26%;7.decreased the level of interleukin-1?(IL-1?)by 6.75%,23.08%,25.81%;8.decreased the level of interleukin-6(IL-6)by 6.62%,27.30%,and 35.10%.(4)Further studies have found that BS-Hex has a protective effect on acute alcoholic liver injury in mice.The experimental results showed that 1.the time of drunkenness within 3 h in mice was 180 min,84 min,142 min,134 min,respectively,among the normal group,the model group,and the BS-Hex group(0.5,1.0 mg/mL),and the drunken rate was 0%,70%,60%,40%;2.real-time blood glucose values of 45 min after drinking were 4.0 mmol/L,7.5 mmol/L,6.1 mmol/L,6.1 mmol/L;3.liver weight coefficient was 4.086%,4.651%,4.291%,4.247%;4.serum ALT levels were 32 U/L,65 U/L,43 U/L,38 U/L;5.serum AST levels were 114 U/L,171 U/L,139 U/L 131 U/L;6.serum amylase(AMS)content were 1094 U/L,1317 U/L,1244 U/L,1110 U/L;7.serum TG content were 0.81 mmol/L,0.95 mmol/L,0.85 mmol/L,0.81 mmol/L.Histopathologically,BS-Hex can prevent hepatocyte enlargement,steatosis and emptying in mice.