The Preparation Of Total Flavonoids In Scorzonera Austriaca Willd. And The Studies On The Hepatic Protective And Anti-hepatitis B Virus Effects

Scorzonera austriaca Willd. is Compositae, Scorzonera genus, perennial herb, andis distributed in the northwest, northeast of China. The whole plant is used as atraditional Chinese medicine, having the effects of detoxification and blood swellingand treating boils, ulcer, snake bites, insect bites, and mastitis.Objective: To extract and purify total flavonoids in Scorzonera austriaca Willd. andstudy the effect of total flavonoids on protecting the liver and anti-hepatitis B virus(HBV), which provides scientific data for developing new drugs that protecting theliver and anti-hepatitis B virus.Methods:(1) Scorzonera austriaca was extracted with ethanol solution immersion atroom temperature, the crude flavonoids was obtained with D101macroporous resinchromatography from the extracting solution, and then the sample was further purifiedwith D4020macroporous resin chromatography to obtain total flavonoids;(2) After Scorzonera total flavonoids was administered ig for7d, ICR mice wereinjected0.1%CCl4by intraperitoneal to induce acute liver injury in2h after finalintragastric administration; ICR mice were injected with BCG2.5mg·kg-1intravenously and after Scorzonera total flavonoids was administered ig for12dlipopolysaccharide were intravenously injected in0.5ug·g-1body weight to induceacute autoimmune liver injury. The blood were collected to detect the enzymes ofaspartate aminotransferase (AST), alanine aminotransferase (ALT), lactatedehydrogenase (LDH) in serum and liver tissue were used as biopsy;(3) Liver cells of Wistar rat were extracted and isolated by orthotopic collagenasedigestion method, and liver cell injury was induced with30mmol·L-1CCl4. Then theliver cells were cultured with total flavonoids of Scorzonera solution for4h. ThenCollecting the supernatant and detecting the concentration of alanine aminotransferase(ALT), aspertate aminotransferase (AST), lactate dehydrogenase (LDH), nitric oxide(NO), superoxide dismutase (SOD) and malondialdehyde (MDA) in liver cells;(4) Culturing HepG2.2.15cell to study the effect of Scorzonera total flavonoids onthe HBsAg, HBeAg and HBV-DNA. Results:(1) D4020macroporous resin for total crude Scorzonera flavonoidsexhibited good adsorption and resolution, is ideal for separating and purifying totalflavonoids.(2) Total flavonoids from Scorzonera can significantly reduce chemical CCl4-inducedliver injury in mice, BCG+LPS-induced immune liver injury and reduce aspartateaminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase(LDH) concentration level in serum(P<0.05,P<0.01).(3) Rat hepatocyte CCl4acute injury model in vitro shows that the optimalconcentration of CCl4-induced liver cell injury in rats was30mmol/L and injury timewas4h; The high, middle, low three dosages group of total flavonoids fromScorzonera can significantly reduce aspartate aminotransferase (ALT), alanineaminotransferase (AST), lactate dehydrogenase (LDH), malondialdehyde (MDA),nitric oxide (NO) concentration level in cell cultural supernatants (P<0.05, P <0.01),while Scorzonera total flavonoids can significantly inhibit the decreasing ofsuperoxide dismutase(SOD)(P<0.05,P<0.01);(4) The high, middle, low three dosages group of total flavonoids can significantlyinhibite the content of HBsAg and HBeAg, and reduce HBV-DNA of HepG2.2.15cellcultural supernatants(P<0.05, P<0.01).The above studies show that Scorzonera total flavonoids can protecte liver andanti-hepatitis B virus. It provides experimental data for the development andutilization of Scorzonera resources and new medicines about hepatoprotective andanti-hepatitis B virus.