| [Background]Our country is a big country of agriculture and animal husbandry is vigorous developing. But recent years, animal bacterial diseases, viral disease and some zoonosis have seriously threatened animal husbandry and people's health. At present the majority medicine and feed additives used in the animal husbandry is antibiotic. In recent years, people realized that using antibiotic makes the drug residue more and more seriously and had already caused the environmental pollution and threatened people's health. Abusing antibiotic easily makes bacteria tolerant to the drug.The Chinese medicinal herbs have the characteristics of lower poisonous and side effect, lower drug residue and difficultly making bacteria tolerant to the drug. So more and more people set their sights on developing the Chinese medicinal herbs resources. Our country possesses the greatest resources of the Chinese medicinal herbs, but plenty of them are waiting to be developed, therefore reasonably developing the resources of the Chinese medicinal herbs is important and has promising future. At present, more and more researcher study on the Chinese medicinal herbs at home and abroad. Research results indicate that Solanum Lyratum Thunb has remarkable antibacterial and antiviral effect and can enhance immunity. But the research of Solanum Lyratum Thunb was not enough, nobody extracted total alkaloids from Solanum Lyratum Thunb and there was no report about the pharmacology of Solanum Lyratum Thunb alkaloids until now.[Objects](1) Studies on the extraction technology of Solanum Lyratum Thunb alkaloid; (2) Appraises the safety of Solanum Lyratum Thunb alkaloid; (3) Measures antibacterial and antiviral effect of Solanum Lyratum Thunb alkaloid; (4) Studies on the influence on immunity of Solanum Lyratum Thunb alkaloid.[Methods](1) Liposoluble alkaloids and water-soluble alkaloids were extracted from Solanum Lyratum Thunb with alcohol and LD50 was measured through reforming Karber way;(2) Antibacterial effect of Solanum Lyratum Thunb alkaloids was studied by cup-plate method and minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of Solanum Lyratum Thunb alkaloids wasmeasured by double dilution tests.(3) Their TD0, TD50, IC50 and TI to chick embryo fibrobast (CEF ) cells were determined. The two kinds of alkaloids were added onto the CEF cells respectively before, after, at the same time the virus or after being put together with virus for 2 hours and their in vitro antiviral effect on NDV was studied.(4) Balb/c mice were divided into four groups: group of control that were injected normal saline(C);group that were injected high(H),moderate(M),low(L) dosage Solanum Lyratum Thunb alkaloids respectively. The four groups were administered for seven days. Phagocytosis of mononuclear macrophage, DTH induced by DNFB, thymus index and spleen index were determined; The mRNA expression level of IL-2,IL-12,IFN- γ and TNF- α were examined by RT-PCR at the 8th day.[Results](1) The extraction result shows that: the content of liposoluble alkaloids and water-soluble alkaloids in the full grass of Solanum Lyratum Thunb is 0.2770% and 1.0095% ; the content of total alkaloids is 1.2865% .(2) The toxicity test indicats: the LD50 of liposoluble alkaloids on mice is 8.46g · kg-1 and the LD50 of water-soluble alkaloids on mice is 8.74g · kg-1.(3) The results shows that water-soluble alkaloids have little antibacterial effect while liposoluble alkaloids have good antibacterial effect against E.coli, Pasteurella multocida, staphylococcus aureus and streptococcus. Its antibacterial diameter is 19.4 ±0.62mm, 20.5 ± 0.57mm, 22.5 ±1.59mm, 22.1±0.82mm.Its antibacterial effect is evident better than Berberine sulpuate ( P<0.01 ) and Lincomycin hydrochloride(P<0.05), is no evident difference comparing with that of Norfloxacin nicotinate(P>0.05). The MIC of Solanum Lyratum Thunb Liposoluble alkaloids to E.coli,Pasteurella multocida, staphylococcus aureus and streptococcus is 6.4mg/ml, 3.2mg/ml, 1.6mg/ml, 3.2mg/ml and MBC is 12.8mg/ml, 12.8mg/ml, 6.4mg/ml, 12.8 mg/ml.(4) The TD0, TD50, IC50 and TI of watersoluble alkaloids is 6.25mg/ml, 11.32mg/ml, 2.48mg/ml and 5.The TD0 , TD50, IC50 and TI of liposoluble alkaloids is 6.25mg/ml, 14.27mg/ml, 1.56mg/ml and 9;The two kinds of alkaloids possess certain antiviral effect, depending on their concentration. Watersoluble alkaloids have better antiviral effect if they are added onto the CEF cells before the virus. Liposoluble alkaloids have better antiviral effect if they are added onto the CEF cells... |
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