| According to relative pharmacology and efficacy record in local Flora about Leucas mollissima Wall.Var.Chinensis Benth, we investigate the anti-bacterial action, anti-inflammatory action, immunization, the effects of removing the phlegm and relieving a cough, and anti-Newcastle disease virus action of its water extract, 95 % alcohol extract, the extracts of alcohol-petroleum ether, chloroform, ethyl acetate, n-butanol and water layer condensate. Then, by comparing the effects of each extract, we can determine the valid target of these drug actions. Above all, our work is a fundamental research for the further purification, isolation and provides theoretical principle to exploitation of Leucas mollissima Wall.Var.Chinensis Benth's.1 Preparation of Leucas mollissima Wall.Var.Chinensis Benth's extracts and the preliminary examination of its chemical compositionObjective: To prepare extracts for our research and get the initial message of the active ingredients. Methodology: A conventional method is used to prepare its water extract and 95 % alcohol extract. We choose petroleum ether, chloroform, Ethyl acetate, and N-butanol as the extracting solvents to separate and purify the chemical constituents in correct order. The system of preliminary examination is applied to study its chemical composition. Results: We get 60.6g frozen drying powder of water extract from 1000g dry herb, the yield is 6.06%; and 141.27g 95% alcohol extract from 2000g dry herb, the yield is 7.06 %; and 30g petroleum ether extract, 17.1g chloroform extract, 20g Ethyl acetate extract, 34.8g N-butanol extract, 23.2g water layer concentrate, their yield is 3.03%, 1.73%, 2.02%, 3.51%, and 2.34%, respectively. The examination shows that this herb contains a variety of materials: steroids or triterpenes, alkaloid, flavonoid, iridoid glycoside, terpenoid lactone, cardiac glycoside, phenols, organic acid, amino acid, peptides, proteins, polysaccharides and amylose, etc. Conclusion: The herb contains a number of active ingredients.2 Researches of antibacterial actionsObjective: To study antibacterial actions of this herb on Staphylococcus aureus, Escherichia coli, Vibrio anguillarum and Bacillus subtilis, get the minimal inhibitory concentration, and determine where the active ingredients are. Methodology: Our study on the herb's antibacterial effects is based on punch method and the Minimal Inhibitory Concentration (hereafter: MIC)of all extracts which are determined by tube dilution method. Results: In the concentration of 200 mg/ml, the extracts of water and 95% alcohol have a significant antibacterial effect on Staphylococcus aureu(sP<0.01), while the water layer condensate has an extremely effect (p<0.01). The Ethyl acetate extract, chloroform extract, N-butanol extract also have the anti-Staphylococcus aureus effect. The water extract (200 mg/ml) only exerts some effects on Vibrio anguillarum but no effect on other 2 sorts of Gram-negative bacteria. However, the 95 % alcohol extract is proved to be functioned on three kinds of Gram-negative bacteria. Through the MIC of extracts on the 4 kinds of bacteria, it's shown that the MIC of extracts on Staphylococcus aureus is much lower than the MIC on the 3 sorts of Gram-negative bacteria, e.g. E. coli. Conclusion: This herb has antibacterial action. The 95% alcohol extract has a little better antibacterial effect than water extract. Our initial inference is that the extracts have more effect on Gram-positive bacteria than on Gram-negative bacteria. The active ingredients of antibacterial upon Staphylococcus aureus are mainly in the water layer condensate, and partly in the extract of chloroform, Ethyl acetate and N-butanol. But the active ingredients of antibacterial on Gram-negative bacteria are in the extract of ethyl acetate.3 Study on antibacterial mechanism through Transmission Electron Microscopy (hearafter: TEM)Objective: To study the herb's antibacterial mechanism on Staphylococcus aureus and Escherichia coli. Methodology: Observe the changes of the 2 kinds of bacteria's structure and characteristics 2h or 6h by TEM dealing with water extract. Results: After initiation of water extract in 200mg/ml concentration, the structure and characteristics of the 2 sorts of bacteria under TEM show significant differences with the normal one. Some cell walls were defected, there was a leakage of contents, and some showed a cavity in the bacteria, etc. Conclusion: This herb has influence on the structure and characteristics of Staphylococcus aureus and Escherichia coli.4 Study on anti-inflammatory actionObjective: To study the herb's anti-inflammatory action and determine where the active ingredients are. Methodology: The experiment is implemented by using anti-swelling of mouse's ear caused by Xylene with the extracts. Results: 700mg/kg (body weight) of 95% alcohol extract , 300mg/kg (body weight) of Ethyl acetate extracts show significant effects on the swelling of mouse's ear (p<0.01). 300mg/kg (body weight) of N-butanol extract, 600mg/kg (body weight) water extract and 300mg/kg (body weight) of Water layer condensate have great effects on the swelling of mouse's ear (p<0.05). Conclusion: This herb has significant anti-inflammatory action. The active ingredients of anti-inflammatory action are mainly in ethyl acetate, and partly in the extracts of n-butanol and water layer.5 Study on immunization actionObjective: To study the herb's immunization action and determine where the active ingredients are. Methodology: Observe the influence on the index of mouse's immune organs such as spleen and thymus to determine its immune effect. Results: Under the given concentration, the 95% alcohol extract and Ethyl acetate extracts increase mouse's spleen index (p<0.05). At the same time, the water extract, 95% alcohol extract and petroleum extract increase mouse's thymus index (p<0.05). Conclusion: This herb can improve the body immunity. The active ingredients of immunization action are mainly in the extracts of Ethyl acetate and petroleum.6 Study on the effects of removing the phlegm and relieving a coughObjective: To study the herb's effects of removing the phlegm and relieving a cough and determine where the active ingredients are. Methodology: The experiment of effect on mouse's cough caused by strong ammonia is applied. Through the method of phenol red in trachea, the effect of anti-sputum is observed. Results: Under the given concentration, its extracts of water, 95%alcohol, ethyl acetate and water layer condensate show significant anti-cough effects. The percentage of relieving a cough (R) is 134 %, 132 %, 135 %, and 134 %, respectively. The water layer condensate and N-butanol extract have significant anti-sputum effect (p<0.01), Water extract and 95 % alcohol extract, Ethyl acetate extract show significant anti-sputum effect (p<0.05). The N-butanol extract and water layer condensate also have some anti-sputum effects. Conclusion: The herb shows remarkable effects of removing the phlegm and relieving a cough. The active ingredients of removing the phlegm effect are mainly in Ethyl acetate extract and water layer condensate. But the active ingredients of relieving a cough effect are mainly in N-butanol extract and water layer condensate, and partly in Ethyl acetate extract7 Study on anti-Newcastle disease virus actionObjective: To study the herb's anti-Newcastle disease virus action and determine where the active ingredients are. Methodology: The anti-Newcastle disease virus action is studied in chick embryo through three ways which are A-way, B-way and C-way. Results: In A-way, only 95% alcohol extract (250mg/ml) has significant (P<0.05) anti-Newcastle disease virus action; In B-way, extracts of 95 % alcohol (250mg/ml) and Ethyl acetate (150mg/ml) show remarkable effect against NDV in embryo (P<0.01), and the water extract (250mg/ml) and chloroform extract(150mg/ml) also play obvious effects(P<0.05); In C-way, the extracts of water, 95 % alcohol and Ethyl acetate show very strong inhibition for NDV in vitro(P<0.01), while the chloroform extract and water layer condensate have some effects(P<0.05). By comparing the 3 ways through statistics, the anti- Newcastle disease virus effects have not much difference. Conclusion: This herb can inhibit NDV in chick embryos. The active ingredients of anti-Newcastle disease virus action are mainly in the extracts of ethyl acetate, chloroform and water layer.
|
PDF Full Text Request