| Objective:1.To study the extraction and enrichment of total phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee and analyzed with qualitaty, quantitaty, structural characteristics, and pharmacological activity of phenylethanoid glycosides.2.To study the different extracted parts of Phlomis younghunsbandii Mukerjee which parts has the hepatoprotective activity on carbon tetrachloride induced liver injury in rats, by observing the different extracted parts of Phlomis younghunsbandii Mukerjee influeneced the levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT) in serum, selected which parts has the hepatoprotective activity.3.To explore the hepatoprotective effects and the mechanism of phenylethanoid glycosides of Phlomis younghunsbandii Mukerjee by macroporous adsorptive resins(EPY-50) on CCl4-induced acute liver injury rats.Method:1.By using D101 macroporous adsorption resin, established the extraction and enrichment of total phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee and analyzed qualitative and quantitative reply on the method of Ultraviolet spectrum.2.70 Wistar rats(female) were fed for a week and randomly divided into 7 groups,normal control group, model control group, bifendate group(100 mg.kg-1), EPY-50(200 mg.kg-1),EPY-25(200 mg.kg-1), water extraction and water elution group. While the normal and model control groups were given distilled water and continuous intragastric administration for 7 days.After administration for 1 hour, rats was intraperitoneal injected with 15%Carbon tetrachloride olive oil solution, the normal control group was intraperitoneal injected with olive oil. After 2 and22 hours, all groups were administered again and observed the physical statation,the blood was draw at 1 h after the last administration from the orbital plexus venosus of rats, standing for 30 minutes, 4500 r·min-1centrifugated for 10 minutes and separated the serum, dilution, detected by biochemical analyzer indexes of aspartate aminotransferase(AST), alanine aminotransferase(ALT).3.60 Wistar rats were randomly divided into 6 groups, normal control group, model control group, bifendate group(100 mg. kg-1) and EPY-50 high-dose, medium-dose and low-dose(400,200, 50 mg.kg-1) groups, the normal and model control groups were given distilled water. After administration for 1 hour, rats was intraperitoneal injected with 15%Carbon tetrachloride olive oil solution, the normal control group was intraperitoneal injected with olive oil. After 2 and 22 hours,all groups were administered again and blood was draw at 1 h after the last administration from the orbital plexus venosus of rats. The levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT) in serum, malondialdehyde(MDA), glutamylcysteinylglycine(GSH), and superoxide dismutase(SOD) in the liver tissue were detected and the index and pathematology of liver were also tested.Result:1.Based on the previous research on the chemical components from Phlomis younghunsbandii Mukerjee, to choose the D101 macroporous resin on the enrichment of components of the water extraction from Phlomis younghunsbandii Mukerjee, scanded in wavelength of 200-400 nm by UV spectrum, the results shown that the 50% ethanol elution and the acteoside as the standard solution have the similar UV scanning spectrum and absorption map in detection wavelength of 330 nm. Meanwhile combined with the results of HPLC chromatogram in previous research, analyzed the three phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee, which concluded that phenylethanoid glycosides was the main components of the 50%ethanol elute by macroporous adsorptive resins from Phlomis younghunsbandii Mukerjee. The contents of total phenylethanoid glycosides increased from 0.16% to 2.41% by D101 macroporous adsorption resin, and the contents increased by 15.06 times.2.The results of activity screening experiments shown that compared with the model group and other groups, total phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee could significantly reduce CCl4 induced chemical liver injury in serum AST and ALT levels of rats( P <0.05, P < 0.01).3.By doing research on hepatoprotective mechanism of phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee, the results shown that compared with normal group, the level of AST, ALT of model group in serum were significantly increased(P < 0.01), liver tissue homogenate of MDA contents increased significantly(P < 0.05), SOD, GSH contents decreased significantly(P< 0.05) liver index significantly increased(P < 0.01), modeling method was a good way to display. Compared to the model control group, EPY-50 with high-dose, medium-dose and low-dose(400, 200, 50 mg.kg-1) could significantly decrease the level of AST, ALT(P <0.05), increase the SOD and GSH, and restricted the MDA in the CCL4-induced liver tissue. The liver index and pathological changes were reversed with 50% ethanol elute by macroporous adsorptive resins(EPY-50) from Phlomis younghunsbandii Mukerjee.Conclusion:1.D101 macroporous resin could be successfully realized the enrichment of total phenylethanoid glycosides.2.The experiment choosen CCl4 as the inducer to induce the acute liver injury model in rats.The experimental results shown that this model could cause the serum levels of AST, ALT significantly increased(P < 0.01) and liver tissue homogenate of MDA contents increased significantly(P < 0.05), SOD, GSH contents decreased significantly(P< 0.05), liver index significantly increased(P < 0.01), modeling method is a good repeatively way to display.The total phenylethanoid glycosides as the main components of the 50% ethanol eluation from Phlomis younghunsbandii Mukerjee, which could significantly reduce the levels of AST and ALT(P <0.01) in rat serum, and the total phenylethanoid glycosides was the main active part with hepatoprotective effect.3.The total phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee on CCl4 induced acute liver injury pathology in rats own an protective effect, the mechanism may be related to the unique pharmacological effects of phenylethanoid glycosides, such as antioxidation,scavenging oxygen free radicals, inhibited the content of MDA as a production of lipid peroxidation, recovered the levels of SOD and GSH, kept oxidation and antioxidation system back to the new balance station and reduced the level of the aminotransferase, may be which is the major reason of phenylethanoid glycosides from Phlomis younghunsbandii Mukerjee has the hepatoprotective effect.This study provides the foundation and the new point for the research of natural medicine of liver protective activity from the medicinal plants from Phlomis younghunsbandii Mukerjee and others. |
PDF Full Text Request