Comparative Studies On The Chemical Components And Pharmacological Effects Of Lonicera Macranthoides Hand-Mazz. And Lonicera Japonica Thunb

Background:As the Lonicera macranthoides Hand-Mazz. was added in2005edition of "Chinese Pharmacopoeia", while the Lonicera japonica Thunb. herbs were divided into Lonicera japonica and Lonicera confusa DC., there exsits much controversy on the difference of chemical elements, clinical application and so on between Lonicera japonica and Lonicera confusa DC.,which is represented by Lonicera macranthoides Hand-Mazz.. Then clinical applications, quality assessment, policy management, market liquidity, economic value, national culture, and other serious questions of these two herbs were caused. So far, there is no systematic study of the main pharmacodynamics of Lonicera macranthoides Hand-Mazz, and no comparative study between Lonicera macranthoides Hand-Mazz. and Lonicera japonicaObjective1. To compare the disparity of main chemical composition in Lonicera macranthoides Hand-Mazz. and Lonicera japonica Thunb..2. To compare the disparity of pharmacological effects of Lonicera macranthoides Hand-Mazz. and Lonicera japonica Thunb.. Methods1. Determination of chemical composition Use an HPLC method for simultaneous determination of chlorogenic aicd, caffeic acid and galuteolin in Fengqiu Lonicera japonica Thunb., Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz. Apply an HPLC method for determination of isochlorogenic acid A. Adopt an HPLC-ELSD method for simultaneous determination of macranthoidin B and dipsacoside B. Apply an UV method for flavonoids.2. Antibacterial activity Plate paper method were applied to test the drug sensitivity.96-well Plate microporous dilution method were applied to assay the MIC and MBC of the following bacterial:Sarcina, Bacillus coli Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus typhi and Bacillus subtilis3. Anti-inflammatory action research(1) An increased capillary permeability model in mice was set up by acetic acid. And mice were divided into6groups, which are normal group, model group(isovolumic purified water), positive group(aspirin0.2g·kg-1), three groups of herbs(10g·kg-1). The inhibition rate was calculated.(2) An acute inflammation reaction model employing xylene as inflammagen to induce mouse ear edema was set up. Grouped as above. The inhibition rate was calculated.(3) Carrageena were employed to induce rat paw swelling. Calculate the inhibition rate by measuring the net value of rat paw swelling in different groups.(4) Rats with granuloma were induce by placing cotton into shoulder cavity. Beside normal group, grouped as above. Calculate the inhibition rate by measuring the net value of tampon granuloma.4. Antipyretic action researchPyretic rats model were set up by the subcutaneous injection of yeast. Grouped as above. Compare the antipyretic action by measuring rats temperature at different time points.5. Immunoregulatory action research(1) The immunosuppressed mice were set up by cyelophosphamide(Cy). Mice were divided into6groups. They are normal group, model group(isovolumic purified water), Cy group(isovolumic purified water), positive group(lentinan4.5mg-kg-1), three groups of herbs(10g·kg-1). Calculate the indexes of spleen and thymus by determine the immune organ weight.(2) The immunosuppressed mice were set up by cyelophosphamide(Cy). Grouped as above.Calculate the clearance index K and the phagocytosis index.(3) Grouped as above, beside normal group. Observing the effect of three kinds of herbs to increase the amount of white cells of rats.Results1. Determination of chemical composition The content of chlorogenic aicd in Fengqiu Lonicera japonica Thunb., Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz. were4.938%,4.575%、9.415%, caffeic acid were0.013%、0.021%、0.021%、galuteolin were0.081%、0.091%、0.038%, isochlorogenic acid A were1.858%、1.9195%、5.129%, and flavonoids were6.815%、6.688%、11.750%. Macranthoidin B and dipsacoside B in Xiushan Lonicera macranthoides Hand-Mazz. were4.352%and2.467%respectively, however, they were not detected in Fengqiu Lonicera japonica Thunb. and Pingyi Lonicera japonica Thunb..2. Antibacterial activity Inhibition zones of Fengqiu Lonicera japonica Thunb., Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz. to Sarcina, Bacillus coli Pseudomonas aeruginosa Staphylococcus aureus, Bacillus typhi and Bacillus subtilis were13.0、4.8、3.2、7.4、8.0、10.0mm,3.2、4.6、7.0、7.2、2.0、6.6mm,8.2、13.4、9.8、10.4、8.2、9.0mm, respectively, MIC were500.0、500.0、250.0、500.0、7.8、250.0mg·mL、1,125.5,125.5、15.7、15.7、15.7、15.7mg·mL-1,7.8、62.8、62.8、31.4、62.8、7.8mg·mL-1, respectively, MBC were500.0、500.0、1000、>1000、1000、500mg·mL-1,>1000、500.0、>1000、>1000、>1000、1000mg·mL-1,250、>1000、>1000、1000、>1000,250mg·mL-1, respectively.3. Anti-inflammatory action research Fengqiu Lonicera japonica Thunb., Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz. had significantly anti-inflammatory action. The inhibition ratios to increased capillary permeability were58.65%、45.19%、55.77%(P<0.001), the maximal inhibition ratios to paw edema were46.67%、47.62%、48.28%(P<0.05), The inhibition ratios to the proliferation of granulation tissue were30.9%、27.2%、37.8%(P<0.001) respectively. Fengqiu Lonicera japonica Thunb. could significantly inhibit ear edema(inhibition ratios was45.1%(P<0.05)), However, Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz.only had inhibition trend(inhibition ratios are38.6%and12.2%(P>0.05).4. Antipyretic action research All these herbs only had antipyretic trend, but did not form significant difference with model group(P>0.05).5. Immunoregulatory action research Fengqiu Lonicera japonica Thunb., Pingyi Lonicera japonica Thunb. and Xiushan Lonicera macranthoides Hand-Mazz. could significantly increase organ indexes(thymus indexes were9.44±1.47,8.43±1.29,9.18±1.16mg·10g-1, spleen indexes were20.16±1.73,19.63±2.42,20.08±2.34mg·10g-1respectively), the clearance index K ((2.03±0.59)x10-2,(1.56±0.74)x10-2>(1.80±0.65)x10-2) and the phagocytosis index (5.26±0.87.4.74±1.00、5.21±1.03)(P<0.001). The amount of white cells of rats be significantly increased by Fengqiu Lonicera japonica Thunb. and Pingyi Lonicera japonica Thunb.(both were9.15±1.65)x109·L-1)(P<0.05), however, Xiushan Lonicera macranthoides Hand-Mazz. only had increasing trend((8.45±1.30)×109·L-1)(P>0.05).Conclusion1. The contents of chlorogenic aicd, isochlorogenic acid A and flavonoids in Xiushan Lonicera macranthoides Hand-Mazz. were more than in the other two kinds of herbs. The contents of caffeic acid in Xiushan Lonicera macranthoides Hand-Mazz. and Pingyi Lonicera japonica Thunb., which were more than in Fengqiu Lonicera japonica Thunb., were equal. The content of galuteolin in Xiushan Lonicera macranthoides Hand-Mazz. was less than in other two kinds of herbs. And macranthoidin B and dipsacoside B were not detected in Fengqiu Lonicera japonica Thunb. and Pingyi Lonicera japonica Thunb.. In the measured seven chemical components, beside galuteolin, the other six chemical components in Xiushan Lonicera macranthoides Hand-Mazz. were more than in the other two kinds of herbs.2. The three kinds of herbs had antibacterial action, anti-inflammatory action and antipyretic trend. They all had immunoregulatory action. And these four aspects of pharmacological effects of the herbs were consistent.