Studies On Antioxidant And Hepaoprotective Effects Of Total Flavonoids From Cirsium Japonicum DC.

Cirsium japonicum DC. is a kind of composite tubuliflorae cynareae conyzoides plants, Its efficacy parts is the aerial parts or root. It is mainly used to treat haematemesis,nose bleeding, hematuri, bloody gonorrhea, flood, leucorrhea, hematochezia, acute appendicitis, large carbuncle, furuncle, high blood pressure, and and so on. this paper studies the process of extraction and purification、the antioxidation and liver protection of flavonoids from Cirsium japonicum DC. This dissertation consists of three chaptersThe first part:the purification and component test research of flavonoids from Cirsium japonicum DC. Including:1. The flavonoids from Cirsium japonicum DC. were extracted by the ultrasonic extraction method with extraction quantity based. As a result of the rhombic experiments, the best extraction technology conditions were that the ethanol concentration was 60%, the solid-liquid ratio was 1:15, the ultrasonic time was 40 min and extracted twice. The extraction rate of flavonoid was 5.98%.2. Two kinds of domestic macroporous resin were used to separate and purificate total flavonoids from Cirsium japonicum DC. in the static adsorption and analytical experiment. The results were that sample solution was 5 mg/mL, the sample volume was 10 mL, the sample rate was 2 mL/min, the useing elution was 60% ethanol, the elution amount was 4BV mL. The content of the flavonoids was about 79.65% which was 1.99 times the pre-purification.3. The main constituents of flavonoids from Cirsium japonicum DC. were pectolinarin and buddleoside by HPLC.The second part:the antioxidant activity research of flavonoids from Cirsium japonicum DC. Including:1. By determining reducing capacity, the antioxidant activity on lipid peroxidation, scavenging activity on DPPH·、-OH、O2、H2O2 and lipid oxidation resistance, antioxidant activities were studied. Ascorbic acid (Vc) and citric acid were compared. The results showed that the reducing power of flavonoid was strong, and the reducing capacity was Vc> flavonoid> citric acid; The removal ability of flavonoid on hydroxyl radicals was stronger, and in the concentration 20-100 μg/mL, the clearance rate reaches 80%, the removal ability of hydroxyl radical was Vc> flavonoid> citric acid; In the between concentration 13-65 μg/mL, The flavonoids had a certain scavenging effect on super oxygen anion. But the role of flavonoids was less than Vc and citric acid; The scavenging effect of flavonoids on DPPH·was stronger than Vc and citric acid. When the concentration of flavonoid was 1 mg/mL, the clearance rate on DPPH · reached 96.38%. The scavenging effect of flavonoids on H2O2 was less than Vc and citric acid; In the lipid peroxide capacity measurement, the IC50 of the flavonoid on lipid antioxidant capacity was 0.33 mg/mL, the ability of flavonoid on lipid oxidation was stronger than Vc and citric acid; The effect of flavonoid on grease antioxidant was obvious and between Vc and citric acid by a quantity-effect relationship.2. The flavonoid of Cirsium japonicum DC. could inhibit erythrocyte hemolysis of mice red blood cell, inhibit the generation of MDA in liver tissue induced by Fe2++H2O2 and the swelling in hepatic mitochondria induced by Vc-Fe2+. The flavonoids of Cirsium japonicum DC. had antioxidation on cell or ganelles of mice in vitro experiment.The third part:Studies on the liver protective and mechanism of flavonoid from Cirsium japonicum DC. obviously on acute liver injury induced by CCI4 in mice.The flavonoids of Cirsium japonicum DC. could protect the liver like Bifendate Pills. The flavonoid of Cirsium japonicum DC. could reduce conspicuously the content of ALT、 AST. ALP in mice serum and the content of MDA liver. It also could improve significantlythe content of SOD、GSH-Px and CAT. The pathological slides showed that in pathology, the volume of was bigger. The liver in model group was fragile. Microscopically, the hepatic lobules were destroyed and the nucleus of liver cells disappeared, the liver cells degenerated, necrosed and lots of inflammatory cells infiltrated around liver. The therapy groups had more significant improvement.