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Initial Studies on the Antibacterical Activity in Vitro and the Safety Evaluation of Propolis Extraction

Author: LiYan
Tutor: JiangShanXiang
School: Nanjing Agricultural College
Course: Veterinary
Keywords: propolis extraction total flavonoids antibacterial evaluate safety
CLC: R285
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract


In domestic and foreign study, some research found that propolis has extensive pharmacologic actions, it known as "purple gold". Propolis can enhance immune function significantly, it also against cancer, regulates blood sugar, antibacterial, and inhibit virus and has other effects. Now it mainly used in health care. In this experiment, we study the toxicity of propolis extraction and in vitro antibacterial activity, in order to provide a theoretical basis for the clinical application in veterinary.1. Extract propolis by cold soaking method and determine content of total flavonoidsIn order to removal the impurity, obtain the propolis extraction, and measure the content of total flavonoid. We use 70% ethanol to extract propolis by hyperacoustic assistant,1:5 of material and liquid ratio, ultrasound 15min, drying the propolis extraction by vacuum oven until the weight of extraction keep constant, and measure the content of total flavonoid by UV spectrophotometer at 415 nm.The result showed that the rate of propolis extract rate is 64.33%, the content of total flavonoid is 18.4%. This extraction method is simple, economic; the method to determination content of total flavonoid is simple, convenient, and accurate enough to determine content of total flavonoid.2. Antibacterial activity of propolis extraction in vitroIn order to evaluate the bacteriostasis effect of propolis extraction, we use Filter paper method to examine inhibition faction of propolis extraction against Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Bordetella by measure the bacteriostasis diameter. The results showed that:The results showed that the sensitivity of Staphylococcus aureus and Bacillus subtilis to propolis extraction are midrange sensitive.The sensitivity of E. coli, Bordetella inhibition are low sensitive.Propolis extraction has a capacity of inhibite the bacterial.3. Evaluation the safety of propolis extraction3.1 Evaluation the general safety of propolis extractionIn order to measure the acute toxicity and sub-chronic toxicity of propolis extraction, we use the maximum tolerated method to determine the propolis extraction LD50, we give propolis extraction to rats by oral administration for 30 day to evaluate the sub-chronic toxicity of propolis extraction. The result found that the propolis extraction LD50>5000 mg/kg, it belong to an actual non-toxic level. In the sub-chronic toxicity test,, the results show that:the realation between the weight and the age is normal; there is no significant difference between organ coefficient of each dose group and negative control group (P>0.05), hematology and blood chemistry examination found that some indexes were significantly different from negative control group (P<0.05), but they changed between the normal value, there is no statistical significance. Pathological examination found that there is no significant pathological changes and necrosis; only found little hyperemia in the kidney, it suggested that there is no sub-chronic toxicity in rats.3.2 Evaluation the inherent safety of propolis extractionObserve the effect to the formation of Micronucleus and the appearance of sperm, in order to evaluate its mutagenic toxicity. In perm structural test, we give propolis for 5d successively, and after the first time administeration for 35d, to put the mice to death, making photos to observe the appearance of sperm;in the micronucleus test, we give propolis extraction 2 times, interval 24h, after the second time administeration for 6h, to put the mice to death, making photos to observe the formation of Micronucleus. The result of the sperm structural test and micronucleus test were negative, propolis extraction has no teratogenic effect.3.3 Stimulus effect to muscle and hemolysis of propolis injectionStimulus effect to muscle and hemolysis of propolis injection were carried out, to evaluate its security of clinical appliation. In Stimulus effect to muscle test,after administeration for 48 h,put the rabbit to death, to observe the reaction orders of quadriceps femors muscle,and make pathological section to observe the injection position of muscle; in the hemolysis test, we observe the the absorption value of urobilinogen and the blood chemistry examination was carried out.The result suggest that:in the stimulus effect test, only found slight hyperemis at the administration position, the summation of reaction progression is 4, pathological examination found that there is no serious necrosis and hyperemia; hemolysis test suggested that the absorption value of urobilinogen between the medication administration team and contral team has no significant difference (P>0.05), blood chemistry examination found that there were no difference between the medication administration team and contral team, the result was negative.It suggested that the propolis injection has no stimulus to muscle, hemolysis in internal.

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