The Stability Of Baicalin-copper Complex And It’s Effect Against Experimental Hepatic Injury In Mice

Baicalin (BA), a flavone glucuronide, is the major active constituent of isolated root of medicinal herb Scutellaria baicalensis Georgi that have many kinds of bioactivities, such as antimicrobial, diuresis and antipyretic effects. Baicalin can coordinate with metal ions because it have hydroxyl group, carbonyl group and many carboxyl group. Our previous research found that baicalin-copper complex (BA-Cu) has stronger antimicrobial activity against Hay bacillus, Salmonella, Candida albicans than baicalin. BA-Cu also has stronger anti-tumor activity against lung cancer cells A549and liver cancer cells HepG2. In this study, we investigated the stability of BA-Cu, the effect of this complex on CCl4-induced hepatic injury in mice, and the anti-inflammatory activity of the complex.BA-Cu dissolved in1%NaOH solution, formulated into30μg/mL solution, respectively at60℃, relative humidity90%±5%,4500lx±500lx glare by UV-visible spectrophotometer at289nm measured the BA-Cu content changes on the0day,5th day and10th day. The results show that high humidity and strong light had no significant effect on BA-Cu (solution content changes within5%), however high temperature had (solution content changes beyond5%).The mice which used for study effect of baicalin-copper complex against hepatic injury were divided into eight groups. The first group was control group. The second group and third group mice were only intramuscular injection BA-Cu solution50,100mg/kg (once daily,12day), respectively. The fourth group mice were intragastric administration with150mg/kg DDB solution (once daily,12day). The fifth group was CCl4group. The sixth group, seventh group and eighth group mice were respectively injected intraperitoneally with CCl4after intramuscular injection with BA-Cu (50,100mg/kg) or intragastric administration with DDB (150mg/kg) once daily for12day. All animal were sacrificed at the16hours after the last administration, and typical acute hepatic injury was observed16hours after CCl4administration. The liver index and histopathology results show that only BA-Cu (all doses,50,100mg/kg) or DDB pretreatment on the mice had no significant difference compared with the control. The CCl4increased liver index, ALT, AST serum levels and decreased SOD level, induced hepatic cord degeneration, and inflammatory infiltration. These changes were altered by preadministration BA-Cu on the mice. These findings suggest that BA-Cu has hepatoprotective effect on CCl4-induced hepatic injury in mice.The contents of Cu in the liver were determined by graphite furnace atomic absorption spectrometry. The1.0g liver samples of the first group, second group and third group mice were collected, after acidification treatment, constant volume with ultrapure water to50mL volumetric flask. The results show that the contents of Cu in the liver of BA-Cu (50,100mg/kg) group were3.8317ppm and5.9374ppm, did not exceed the limit of the LD50(110ppm, Chinese Academy of Sciences report). Hence, BA-Cu did not lead to heavy metal poisoning.The mice which used for research effect of baicalin-copper complex on auricular swelling model were divided into four groups. The first group was control group. The second group and third group mice were only intramuscular injection BA-Cu solution50,100mg/kg (once daily,12day), respectively. The fourth group mice (positive control group) were intragastric administration with20mg/kg dexamethasone solution, once daily for12day. After the last administration,30μL xylene was used for induced each mouse’s right ear swelling, left ear was not used. All animal were sacrificed at the1hour after the xylene treatment, with a punch to remove the ear piece at the same site of two ears, weighed pieces, then figure out the weight difference of left and right ear piece, and inhibition rate (%). The results show that BA-Cu significantly attenuated xylene-induced mice auricular swelling, high dose group has more remarkable anti-inflammation effect (inhibition rate were59%).These test results show that BA-Cu has markedly hepatoprotective effect and anti-inflammation effect. BA-Cu is stable at room temperature, easy to preserving.