Rutin, Danshensu, In Vitro Studies, Anti-oxidation

The oxygen molecule might produce highly reactive oxygen species (ROS) by some exogenous factors and endogenous metabolic processes in human body. They can bring many kinds of illness, such as cardiovascular disease, senile dementia, tumor and sugar diabetes and so on, when the free radical in body is imbalance caused by some factors. Numerous researches found that taking preventions, especially taking antioxidant or free radical scavenger reasonably can effectively keep above diseases from happening. In the last century, the synthetic antioxidants such as butylated hydroxyltoluene (BHT) and butylated hydroxyanisole (BHA) were widely used in food industry. However, a plenty of experimental data indicated that these synthetics could cause many diseases and cancer. So nature antioxidant has got people's favor again, because of its extensive source, strong antioxidant activity and excellent security. Therefore, the development of effective and safe natural antioxidants becomes one of the main topics in the field of food chemistry.Rutin is widely distributed in vegetables and fruits, and has been widely used in Chinese traditional medicine for anti-inflammatory and vasoactive, antitumor, antibacterial, antiviral and antiprotozoal properties. And Salvia miltorrhiza Bunge is a traditional Chinese medicine and has long been used for treating liver and heart diseases in China. The main hydrophilic active component of Salvia miltorrhiza Bunge is Salvianic acid A, which has many pharmacological effects which includes protection of cardiac muscle, dilatation of blood vessel, inhibition of arteriosclerosis and thrombus, improvement of microcirculation, regulation of restoration and regeneration of tissue, antisepsis and anti-inflammation and so on. But the reports of rutin and Salvianic acid A about free radical scavenging activity and antioxidant effect in mitochondria are even less.This paper introduced the general situation and pharmacologic function of rutin and Salvianic acid A. We hope that through serial researches, we can provide references and theoretical foundation for the exploiture and utilization of them.In this paper, the antioxidant activities of the rutin and salvianic acid A were investigated by various established in vitro systems, such as total antioxidant activity, reducing power, hydrogen peroxides scavenging activity, superoxide radical scavenging assay, DPPH radical scavenging assay, lipid peroxidation assay which using egg yolk as the lipid-rich source. Total antioxidant capacity was determined by the assay based on the reduction of Mo(VI)-Mo(V) by the sample and subsequent formation of a green phosphate/Mo(V) complex. The results show that rutin and salvianic acid A have strong antioxidant capacity. Rutin has strong scavenging free radical capacity and the radical-scavenging effects were in a dose-dependent manner, which can effectively remove DPPH·,O₂^-·and H₂O₂, and the IC₅₀ were 26.6μg/mL, 0.27mg/mL and 0.44mg/mL, respectively. In addition, rutin has effective inhibition of lipid peroxidation with IC₅₀ of 0.37mg/mL. At all these assaies, salvianic acid A has stronger capacity than rutin. The IC50 of removing DPPH·,O₂^-·, H₂O₂ and inhibition of lipid peroxidation are 7.0μg/mL, 0.098mg/mL, 0.33mg/mL and 0.092mg/mL.In the present study, we also evaluated the abilities of rutin and salvianic acid A to protect the liver mitochondria. The ability of scavenging O₂^-·generated from mitochondria, the inhibitory effects of rutin on lipid peroxidation of mitochondrial membranes, mitochondrial membrane permeability transition, inhibitory protein carbonyl oxidation and protection ATPase of rutin and salvianic acid A were evaluated in rat liver mitochondria. We found that rutin and salvianic acid A could effectively scavenge superoxide anions in a dose-dependent mode with IC50 of 0.086mg/mL and 0.014mg/mL. Free radicals can initiate lipid peroxidation that may cause tissue damage. Our results show that rutin and salvianic acid A significantly can inhibit the formation of protein carbonyls in Fe²⁺/Vc-initiation. Especially, 0.01mg/mL of salvianic acid A prevents the Fe²⁺/Vc -induced protein oxidation by 77.3±0.77%, and that of 0.25mg/mL of rutin is 58.8±0.86%. From the results, we can see that rutin and salvianic acid A at all concentration significantly protect the activities of ATPase.Rutin and salvianic acid A have shown to inhibit the rat liver mitochondrial swelling and the oxidation of carbonyl groups to ensure that the mitochondrial membrane osmotic pressure exterior and interior. It also protected mitochondria membranes by scavenging superoxide anions and scavenging lipid free radicals as well as maintaining ATPase activity obviously. Mitochondrial membrane has important defensive function to protect cells. So they can protect mitochondria well.In conclusion, rutin and salvianic acid A have significant antioxidant activity in vitro and effective protection of liver mitochondrial, which suggested the pharmacological potential of them against pathological processes related to oxidative stress. Therefore, it is suggested that further work could be performed on the mechanism of the antioxidant activity and the clinical management of rutin and salvianic acid A as antioxidant supplements due to its potent antioxidative effect.