The Antioxidant And Anti-hypoxia Effects Of Norepinephrine Wogonin

Objective:1. Designed and synthesized Norepinephrine Wogonin and identified its structure.2. Established eight kinds of antioxidant activity measurement methods and investigated the antioxidant activity of Norepinephrine Wogonin.3. Established PC12cells hypoxia model and study the protection from the effects of Norepinephrine Wogonin on the hypoxic cells, screening the best Norepinephrine best drug concentration of Norepinephrine Wogonin in PC12cells under hypoxia, study the possible protective mechanism of Norepinephrine Wogonin in hypoxic cells.Methods:1. Norepinephrine Wogonin was gained after five steps using chrysin as starting material.2. The antioxidant effects of Norepinephrine Wogonin was studied using DPPH\, ABTS+·,O2-··OH, NO Scavenging assy, reducing power, T-AOC and FRAP methods.3. Establish PC12cells hypoxia model, First we can detect the activity of hypoxia cell was detected by MTT assay, LDH content, ATP content, ROS contente, MD A content, SOD activity, T-AOC content in PC12cells hyopxia mode; Then we study the protection of Norepinephrine Wogonin by MTT assay, HE staining morphology, LDH content, ATP content, ROS content, MDA content, SOD activity, T-AOC content, also detect intracellular calcium, use Real-time PCR analysis HIF-1α,VEGF, Cytochrome C, Bcl-2, Caspase-3mRNA expression levels, and use Western-blot detect HIF-la,VEGF, Cytochrome C, Caspase-3proteins expression.Results:1Norepinephrine Wogonin was got through a series of reactions using chrysin as start material with a total yield of57%and high pruity greater than99.0%.2The results of eight kinds of classic antioxidant erperiments showed that Norepinephrine Wogonin was of good clear DPPH·,ABTS+, O2-·,·OH, NO Scavenging ability, certain effect on reducing power, and good total antioxidant and iron reduction/antioxidant.3The results of PC12cells hypoxia models showed that hypoxia could inhibit cell growth, increase medium LDH levels, reduce ATP content, increase intracellular ROS levels, increase the accumulation of MDA, reduce intracellular SOD, T-AOC activity; Norepinephrine Wogonin which at the final concentration of1×10-6mol·L-1, it could increase cell survival (P<0.01), decrease medium LDH levels (P<0.01), increase ATP levels, reduce intracellular ROS levels (P<0.01), reduce the accumulation of MDA (P<0.01), increase intracellular SOD, T-AOC (P<0.01) activity, decrease levels of intracellular calcium (P<0.01), reduce calcium influx, significantly increase Bcl-2mRNA expression (P<0.01), reduce HIF-la, VEGF, Cytochrome C, Caspase-3mRNA and protein expression levels (P<0.01), increase the cell antioxidant, protect the cell membranes and reduce apoptosis.Conclusion:1. Synthesize Norepinephrine Wogonin and the method is simple, of low cost and high purity.2. Norepinephrine Wogonin has a good antioxidant activity,which has a good clear DPPH·,ABTS+,O2-·,·OH, NO Scavenging ability, certain effect on reducing power, and good total antioxidant and iron reduction/antioxidant.3. Norepinephrine Wogonin shown protective effect on PC12cells in hypoxia. The possible mechanism was that it could improve cells activity, scavenging intracellular excessive ROS, increase intracellular antioxidant capacity, and reduce the accumulation of lipid peroxidation products, increased intracellular the ATP content, increased mitochondrial membrane potential, reduce calcium influx, reduce cytochrome C gene and protein expression in cells, thereby protecting the mitochondria, the integrity of cell membranes, and reducing apoptosis. further research needs specific mechanisms.