| Oxidative stress is a frequent problem in livestock breeding,it is an important reason for the decline of the resistance of livestock.Serious oxidative stress reaction may even endanger the life of livestock and cause huge economic losses to the breeding industry.Although synthetic antioxidants have good antioxidant properties,in recent years,pharmacological studies on synthetic antioxidants have found that they have some toxic and side effects.It is easy to produce residues in the body,which is a hindrance to the ecological,healthy and sustainable development of the livestock and poultry industry,so the research of antioxidant components of medicinal plants,and development of natural antioxidants has become an important research direction.Natural flavonoids as an antioxidant substance with good performance in animal disease prevention care has been well received by the researchers,flavonoids have a variety of biological activity,the antioxidant properties of flavonoids were applied to livestock health and fitness,also need further in-depth study of the antioxidant mechanism in animal body,cellular and molecular level.Acanthopanax senticosus as a cheap and convenient traditional Chinese medicinal material,has the functions of strengthening muscles and bones,reinforcing function of the liver and kidney,etc.According to the research,Acanthopanax senticosus can improve the immunity of livestock and poultry,resisting inflammation,improving the flora and structure of intestinal tract,reduce the mortality of young animals.In order to study the antioxidant ability of Acanthopanax senticosus flavonoids,this experiments was to discuss the extraction process of Acanthopanax senticosus flavonoids,and the quality standards of Acanthopanax senticosus flavonoids was established.The antioxidant properties of flavonoids from Acanthopanax senticosus were studied on the model of chicken embryo fibroblast（CEF）induced damage by hydrogen peroxide that provide theoretical basis for clinical application.The results are as follows:1.Establishment of a method for the determination of flavonoids in the stem of Acanthopanax senticosus:Rutin was used as standard sample,The standard curve was established by using sodium nitrite-aluminium nitrate-sodium hydroxide chromogenic method.The results showed that the regression equation of the standard curve is:A=0.0115c-0.0064,R2=0.9995,Precision test,stability test,repeatability test and added sample recovery test all accord with the standard of pharmacopoeia.2.Response surface method was used to optimize ultrasonic assisted ethanol extraction process of flavonoids from stem bark of Acanthopanax senticosus:Ultrasonic assisted ethanol extraction technology was adopted,determine the selection factors of response surface through the single factor experiment.The optimum ultrasonic assisted extraction process was optimized by Box-Behnken Design response surface.The results showed that the optimum extraction process of flavonoids from stems of Acanthopanax senticosus was as follows:extraction temperature 65℃,extraction power 399W,extraction time 45 min,and the maximum predicted extraction yield was3.756%.According to the optimum extraction process,three verification testing were carried out according to the optimum extraction conditions,the average extraction yield was 3.72%.3.Evaluation of Antioxidant ability of flavonoids from the stem bark of Acanthopanax senticosus in vitro:The reducing power,scavenging rate of DPPH、superoxide anion radical、hydroxyl radical and ABTS free radical were determined with using VC as control by test,the antioxidant activity was measured in vitro.The results showed that flavonoids of Acanthopanax senticosus had good antioxidant activity,the IC500 value of clearance ratio of Acanthopanax senticosusflavonoidsforthe DPPH radical,superoxide anion,hydrogen peroxide,hydroxyl radical and ABTS radical,were as follows:2.305μg/mL、34.696μg/m L、5.489μg/mL、2.970μg/mL、9.256μg/m L。4.Preprotective effect of flavone from the Acanthopanax senticosus stem for the CEF cells are induced by hydrogen peroxide that expressed oxidative stress:The tests has been cultured chicken embryo fibroblasts（CEF）,determined the maximum safe concentration of flavonoids in the stem of Acanthopanax senticosus and hydrogen peroxide;CEF cell model induced damaged by hydrogen peroxide was established,the preprotective effect of flavonoids on oxidative stress induced by hydrogen peroxide was measured,and the activity of antioxidant enzymes in the cells were also detected.The results showed that the oxidative stress model of CEF cells induced by hydrogen peroxide was successfully established,Flavonoids could pre-protect CEF cells from stress,increase the total antioxidant capacity and the contents of peroxidase SOD and GSH-Px,decrease the content of MDA,suggesting that flavonoids may play an anti-free radical effect by regulating the content of antioxidant enzymes. |
