Poplar Bud Extract Extraction Technology And Active Research

Poplar is one of the most widely planting tree species in our country,the poplar planting area is about 8 million hectares in our country,annually in December to next February is the season of poplar sprouting.the new poplar buds contain rich pectin,some scholars’ study found that poplar bud extract is rich in flavonoids,phenols and terpenes,which generally has a good antioxidant effect,can effectively remove free radicals in the body.Modern research has found that the oxygen free radicals involved in the vast majority of human diseases,such as cardiovascular disease,cancer,inflammatory response and so on.Object:Optimizing the extraction process of the poplar buds to optimize extraction rate,and fill the domestic blank in this field.Trying to establish HPLC fingerprint of poplar bud extract and improving the quality control standards.To determine the in vivo and in vitro antioxidant activity of the optimized poplar bud extract,and research on the capability of removing the free radicals.Preliminary study on the apoptosis mechanism of the poplar bud extract on human gastric cancer SGC-7901cells,in order to laying a foundation for the pharmaceutical industry of poplar bud.Method:1.Ultraviolet-visible spectrophotometry is a detection method,through this to do the single factor test and than determine orthogonal experiment conditions,through the orthogonal experiment investigation after ethanol volume fraction(%),extraction time(min)and the material liquid ratio on poplar buds of flavone extraction process,determine the optimal extraction process.2.The HPLC method was adopted to detect the samples.The separation was performed on Merck C18(4.6x250 nm)with the mobile phase consisted of methanol and 0.4%H3PO4 at a 1mL/min flow rate,a 320nm detection wavelength,a column temperature of 30℃and 10μl,of injection volume.The Similarity Evaluation System for Chromatographic Fingerprint of traditional Chinese medicine(2004 A)was used to get the fingerprint of poplar bud extract,check out the coffee acid and the others.3.By using the DPPH method,the autoxidation of pyrogallol method and CuS04-Phen-Vc-H202 chemiluminescence system to measure its in vitro antioxidant activity.Besides,the CuS04-Phen-VC-H202 chemiluminescence system was adopted to study its protective effect on DNA damaged by · OH.4.The experimental mice were randomly divided into 5 groups,the different doses of poplar bud extract high,medium and low groups(40 mg/kg,20 mg/kg,and 10 mg/kg),VC positive group(10 mg/kg)and blank group,Continue gavage in them for 14 days,orbital blood next day after the last gavage,4 ℃,3500 RPM centrifugal for 10 minutes,took the supernatant,determined the content of MDA,SOD,T-AOC and GSH-Px.Anatomy of the mice,took the liver,spleen,thymus out,weighing them respectively,calculating the thymus index and spleen index,the liver tissue of slurry preparation,the liver tissue of determination of serum MDA,SOD,T AOC and gsh-px levels.Comprehensive results determined the poplar bud extract antioxidant activity levels in the body.5.MTT method was used to detect the inhibiting effect of the poplar bud extract on the SGC-7901 cells,a human lower differentiated gastric adenocarcinoma cell line.Flow cytometry was used to study the apoptosis of the SGC-7901 cells after medical intervention after 24/28 h,and western blot method was used to detect the protein expression of Bax,Bcl-2 and Caspase-3 in the SGC-7901 cells.Results:1.On the basis of the orthogonal test,the best conditions of extraction is 10 times 55%ethanol solution and time for 40min.2.Using optimized extraction method got fingerprints of 12 batches,this method can make effective components from the poplar trees bud gets better separation,the final result shows that poplar bud extract contains 13 common peak,which determine the five kinds of caffeic acid,P-coumaric acid,ferulic acid,celery and chrysin,for further study its specific composition laid the early foundation.3.The inhibitory rates of DPPH radicals was 38.1%-94%when the concentration was between 0.05-0.8 mg/mL.However,the eliminating ratio of O2-increases along with the increase of the amount of matter and changed between 37.4%-44.2%when the concentration was at 0.1 mg/mL.The · OH were scavenged 40.8%-60%when the concentration was between 0.05-0.8 mg/mL.The results showed that the poplar bud extract with different concentration possesses a favorable protection effect on DNA oxidant damage especially concentration at 0.4 mg/mL.4.Medium and high concentrations of poplar bud extract could significantly improve the level of immunity in mice,increasing the mice spleen index and thymus index,also increasing SOD,gsh-px and T-AOC level in the body,and decrease MDA content,showing a good immune enhancement and antioxidant activity in the body.5.The MTT results showed that the poplar bud extract possesses a prominent inhibitory effect on the proliferation of the SEC-7901 cells in a time and dose dependent manner.The flow cytometry results implicated that the poplar bud extract could remarkably promote the apoptosis of the SEC-7901 cells which is positively related to duration time.Western blot results showed that after 48 h,poplar sprout extract SGC-7901 cells high Bax gene expression,The Bcl-2 gene expression increased with concentration and Caspase-3 gene no expression.