Preliminary Study On Antioxidant Effect Of Canarium Pimela Leaves Extract In Vivo And In Vitro

Background:China's chronic disease deaths accounted for 86.6 percent of the total death toll.A large number of studies have shown that antioxidant stress is an important factor to control the occurrence and development of chronic diseases.Antioxidants can be used to alleviate the occurrence of oxidative stress.At present,the safety of synthetic antioxidants has been widely questioned,so finding safe and effective natural antioxidants is an urgent problem to be solved.The Canarium pimela leaves can be harvested throughout the year and are rich in resources.The study found that it has strong scavenging effects on oxygen free radicals and anti-aging,but its efficacy and mechanism have not been studied in depth.Therefore,it is of great economic value and application value to study the antioxidant effect of Canarium pimela leaves and infer its mechanism.Objective:In this study,the constituents,antioxidant activity of Canarium pimela leaves in vitro and in vivo were discussed.The aim is to provide scientific basis for the development of Canarium pimela leaves and the study of natural antioxidants.Methods:1.?1?Determined the contents of total flavonoids,polyphenols,saponins and polysaccharides in different extracts of Canarium pimela leaves by colorimetry;?2?To evaluate its antioxidant activities by measuring the free radical scavenging power,reducing power and anti-lipid peroxidation;?3?Correlation between content and antioxidant capacity was analyzed by Perman.2.?1?Taking the content of total flavonoids as an index,combined with orthogonal experiment,the optimum extraction technology was determined;?2?After purification,the total flavonoids of Canarium pimela leaves?TFCPL?,were obtained and their antioxidant activities were studied.3.?1?The oxidative damage model mice were established by injection of D-galactose.The liver,lung and other organs of the mice were stained with HE and observed.The activities of T-AOC?GSH-PX?SOD?TNF-??iNOS?NO?CAT in serum,liver and brain of mice were determined by ultraviolet spectrophotometry and enzyme plate method.?2?C₁₈8 column was used,with gradient elution of ortho-phosphoric acid and methanol as mobile phase.Based on the literature report and the comparative analysis of the HPLC chromatograms between the blank serum and the drug-containing serum and the reference substance after oral administration of TFCPL,the blood-entering components were preliminarily inferred.Results:1.?1?the content of total flavonoids in CEE was the highest,which was significantly different from other extracts?P<0.01,P<0.05?;?2?CEE had obvious scavenging effect on DPPH·?16.31ug/ml?,ABTS⁺·?0.240mg/ml?,·OH?0.170mg/ml?,reducing power?0.221±0.005?and lipid peroxidation?85.84±0.35%?;?3?There was a significant correlation?P<0.01?between its content and antioxidant activity.2.?1?The optimum extraction conditions were as follows:ethanol with 50%volume fraction,liquid-to-material ratio of 1/20,reflux extraction time of 1.5 h,extraction time once;?2?The antioxidant activity(IC₅₀=0.065mg/mL)of TFCPL was more persistent than vitamin C.3.?1?There was no significant effect of TFCPL on the body weight of mice?P>0.05?.TFCPL group had different degrees of inhibition of hepatic pneumonic degeneration and inflammatory infiltration of the lungs,and so on.Compared with the model group,the levels of GSH-Px,SOD in serum and the contents of MDA,TNF-?and iNOS in each TFCPL group were significantly higher than those in the model group?P<0.01,P<0.05?.The activities of antioxidant enzymes such as T-AOC?GSH-Px and CAT in liver were significantly increased?P<0.01,P<0.05?.The content of NO decreased significantly?P<0.05?.The contents of MDA in brain tissue were significantly decreased?P<0.01,P<0.05?;It was found that four kinds of components in elemite leaves might be absorbed into blood directly,suggesting that these components may play a role in efficacy.Conclusion:1.The antioxidant activity of CEE was the best,the content of total flavonoids was the highest.2.The antioxidation capacity of purified TFCPL was higher than that of BHT and the total antioxidant capacity was higher than that of V_C.3.TFCPL could improve the pathological changes of the organs and increase the activity of the enzymes in the organs of D-galactose-induced oxidative injury mice.The mechanism may be related to the increase of antioxidant enzyme activity.4.It is speculated that the blood-entering components of TFCPL may be vitexin,isovitexin,hyperin and quercetin,which need to be further verified.