Off-line Antioxidant Analysis And Anti-hypoxia Activity Of Anthocyanins From The Fruits Of Lycium Ruthenicum Murr

The Qinghai-Tibet Plateau is the highest and most populated plateau in the world with unique environment.The special environment on the plateau such as low pressure,low oxygen,cold and strong ultraviolet rays that have an ongoing impact on the life and health of personnel working and traveling in Tibet,and national defense construction at all times.Medical research that can cure acute and chronic plateau diseases such as high altitude cerebral edema,pulmonary edema,and polycythemia is very important for local economic construction and national security.In recent years,drug research based on the mechanism of high altitude hypoxia injury such as peroxidation and inflammatory outbreak was in the ascendant.As a main component from the fruits of Lycium ruthenicum Murr,anthocyanins were analyzed in chemical and pharmacology due to the wide application in traditional Tibetan medicine.However,there were few studies on the anti-hypoxia activity of anthocyanins because the research basis for hypoxia injury medicine was deficient.In this work,High Performance Liquid Chromatography（HPLC）was used to analyze each anthocyanin components from the fruits of Lycium ruthenicum Murr,including detecting the content and analyzing the ability to scavenge 1,1-diphenyl-2-picrylhydrazyl（DPPH·）free radicals.Anti-hypoxia function of anthocyanins from the fruits of Lycium ruthenicum Murr was evaluated by using normobaric hypoxia and sodium nitrite hypoxia mice model and hypobaric hypoxia rat model.The main results are as follows:（1）The content of each component in the anthocyanins from the fruits of Lycium ruthenicum Murr extract was determined using HPLC,and the gradient elution with acetonitrile-0.1%phosphoric acid aqueous solution was used.The results showed that the six kinds of anthocyanidin monomers able to sustain the baseline separation,and had good peak shape.The total content of anthocyanins in the fruits Lycium ruthenicum Murr extract was 231.95 mg·g^-1,of which Petundin had the highest pigment content,which was 213.09 mg·g^-1.（2）The IC₅₀ value of anthocyanins from the fruits of Lycium ruthenicum Murr for scavenging DPPH·radical was 0.0138 mg·m L^-1.The establishment of DPPH-HPLC off-line antioxidant analysis method confirmed that each component of anthocyanins from the fruits of Lycium ruthenicum Murr had good antioxidant capabilities.The oxidative capacity aids in the analysis of antioxidant active compounds on a technical level.（3）The administration method and doses of anthocyanins from the fruits of Lycium ruthenicum Murr were screened out by normobaric hypoxia experiment and sodium nitrite hypoxia experiment.The results showed that the anti-hypoxia effect of anthocyanins from the fruits of Lycium ruthenicum Murr by gavage was not noticeable,but the effect of intraperitoneal injection of anthocyanins from the fruits of Lycium ruthenicum Murr at low,medium and high doses(50,100,and 150 mg·kg^-1)had significant effects.The prolongation rates of the anti-close normobaric experiment were 26.24%,51.83%,and 60.31%,respectively,and the prolongation rates of hypoxia experiment by sodium nitrite were 5.92%,10.81%,and 25.30%,respectively,showing a dose-dependent manner.（4）Analysis of the protective effects of low,medium and high doses(35,70,105 mg·kg^-1)of anthocyanins from the fruits of Lycium ruthenicum Murr and positive control acetazolamide(250 mg·kg^-1)on cerebral and pulmonary edema in rats under hypobaric hypoxic environment were studied.The findings reveal that anthocyanins from the fruits of Lycium ruthenicum Murr could decrease the water content of brain and lung tissue in rats with simulated high altitude injury,and as well as the pathological damage of rat brain and lung tissues.Furthermore,measurement results of oxidative stress and inflammation-related indicators in rat brain and lung tissues showed that anthocyanins from the fruits of Lycium ruthenicum Murr can reduce the content of MDA and ROS in the homogenate of brain and lung tissue of hypoxic rats,increase the amount of GSH and the activities of T-SOD,CAT and GSH-PX,and reduce the content of IL-1β,VEGF,TNF-α,IL-6.Finally,anthocyanins from the fruits of Lycium ruthenicum Murr may have an anti-hypoxia effect by scavenging free radicals and reducing inflammation,suggesting that anthocyanins could be used to develop anti-hypoxia drugs.