山莓叶形态解剖与抗菌成分及毒副作用研究
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摘要
山莓(Rubus corchorifolius L.f.)是蔷薇科悬钩子属(Rubus L.)落叶灌木植物,资源极其丰富。山莓叶具有抗腹泻作用,已有悠久的应用历史。本文对山莓叶的形态结构、功能成分及其抗菌效果与毒副作用等方面进行了系统而较深入的研究。研究结果为山莓叶作为药物饲料添加剂以替代抗生素在饲料中应用提供了依据。主要研究结果如下:
1、山莓为阳性植物,叶为典型的背腹型叶,并有阳叶和阴叶之分,其结构存在明显差异。
2、不同月份山莓叶粗提物的提取率差异显著:鞣质、香豆素和黄酮类物质的含量存在地域差异;从山莓叶挥发油中分离出49个组分,鉴定出45个,其中1,2,4-三甲氧基丁烷达52.1%;分析中发现鞣质类中含没食子酸,脂溶性香豆素类提取物中含香豆素,分离出特征性黄酮化合物山奈酚-3-O-β-D-(6”-对羟基桂皮酰基)-葡萄糖苷在山莓叶中首次发现。
3、不同月份粗提物都有良好的抑菌效果,但对供试5种菌表现出一定的差异性:以7月的作用最强,其中鞣质对痢疾志贺氏菌的抑菌作用最强,对大肠杆菌次之,且都优于复方黄连素;50 mg/mL的香豆素抑菌效果除对大肠杆菌作用较弱外,对其它细菌效果较强(强于复方黄连素);黄酮都有很好的抑菌效果,对金黄色葡萄球菌的作用最强;挥发油具有很强的杀菌作用,对痢疾志贺氏菌的作用最强,大肠杆菌次之,0.5%浓度时,仍稍优于0.4%的黄连素。
4、碳廓清试验表明,鞣质提取物组和高剂量的香豆素组对小鼠的胸腺指数有影响,其余组份与对照没有明显差异;黄酮组校正吞噬指数显著高于其它组,说明山莓黄酮可提高小鼠巨噬细胞的吞噬活性,但不同剂量之间没有明显的差异:但香豆素、鞣质和山莓粗提物对小鼠的巨噬细胞的吞噬活性没有影响,其组织形态结构未发现病理变化,无毒副作用。
The Rubus corchorifolius L.f. is a wild deciduous shrub of rose family (Rosaceae), which is abundant in the wild. The effect of anti-diarrhea of its leaf has been known for a long time. In order to establish the theory basis of application of R. corchorifolius as a feed additive instead of antibiotics, the morphological and anatomical characteristics, antibacterial effects and toxic side effects in its functional compositions have been systematicly studied. The results are as follows:
1. R. corchorifolius is a sun plant with bifacial leaf which can be divided into sun-leaf and shade-leaf, there are obviously differences in structure between the two leave types.
2. There are great differences in the crude extraction rates of the leaves of if. corchorifolius in different months. The components, such as tannin, coumarin and flavone, vary in materials tested from different region. 49 compositions have been separated from its leaf essential oil; and 45 of them were identified. The content of (R) Butane. 1.2.4-trimethoxy reached 52.1%. Gallic acid was found in tannin, the kampfreol-3-O-β-D-6"-P- hydroxycoumaroyl-glucosie, one of flavonoid compounds, was firstly separated in the plant.
3. The leaf rude extraction in different months have different antibacterial activities to the 5 bacterium tested which in July is the best. Tannin is the best on inhibiting Shigella dusenterlea, the second on inhibiting Escherichia coli. Both are better than that of berberine. The antibacterial activity (except inhibiting Escherichia coli) of coumarin (50mg/mL) was better than that of berberine. Flavones have strong antibacterial activities, especially inhibiting Staphylococcus aureu. Essential oil has a strong sterilization effect; inhibiting Shigella dusenterlea is the best, inhibiting Escherichia coli the second. Sterilization effect of essential oil is a little better than that of berberine at the concentration of 0.5%.
4. There were no significant differences of liver index and spleen index between experimental group and control group, except the extract of tannin and coumarin of high dosage. The corrected phagocytic index (a value) of the flavone group was significantly higher than that of other groups (P≤5). It shows that the flavone in R. corchorifolius can enhance macrophage phagocytic activity. But there were no significant differences in phagocytic activity among each dosage of flavone. And there were no obvious differences in a value among the rest three groups and the CK. There were no pathological changes found by the observation of paraffin section of spleen, liver and kidney. It shows that the application of the compounds tested are safety to animals.
1、山莓为阳性植物,叶为典型的背腹型叶,并有阳叶和阴叶之分,其结构存在明显差异。
2、不同月份山莓叶粗提物的提取率差异显著:鞣质、香豆素和黄酮类物质的含量存在地域差异;从山莓叶挥发油中分离出49个组分,鉴定出45个,其中1,2,4-三甲氧基丁烷达52.1%;分析中发现鞣质类中含没食子酸,脂溶性香豆素类提取物中含香豆素,分离出特征性黄酮化合物山奈酚-3-O-β-D-(6”-对羟基桂皮酰基)-葡萄糖苷在山莓叶中首次发现。
3、不同月份粗提物都有良好的抑菌效果,但对供试5种菌表现出一定的差异性:以7月的作用最强,其中鞣质对痢疾志贺氏菌的抑菌作用最强,对大肠杆菌次之,且都优于复方黄连素;50 mg/mL的香豆素抑菌效果除对大肠杆菌作用较弱外,对其它细菌效果较强(强于复方黄连素);黄酮都有很好的抑菌效果,对金黄色葡萄球菌的作用最强;挥发油具有很强的杀菌作用,对痢疾志贺氏菌的作用最强,大肠杆菌次之,0.5%浓度时,仍稍优于0.4%的黄连素。
4、碳廓清试验表明,鞣质提取物组和高剂量的香豆素组对小鼠的胸腺指数有影响,其余组份与对照没有明显差异;黄酮组校正吞噬指数显著高于其它组,说明山莓黄酮可提高小鼠巨噬细胞的吞噬活性,但不同剂量之间没有明显的差异:但香豆素、鞣质和山莓粗提物对小鼠的巨噬细胞的吞噬活性没有影响,其组织形态结构未发现病理变化,无毒副作用。
The Rubus corchorifolius L.f. is a wild deciduous shrub of rose family (Rosaceae), which is abundant in the wild. The effect of anti-diarrhea of its leaf has been known for a long time. In order to establish the theory basis of application of R. corchorifolius as a feed additive instead of antibiotics, the morphological and anatomical characteristics, antibacterial effects and toxic side effects in its functional compositions have been systematicly studied. The results are as follows:
1. R. corchorifolius is a sun plant with bifacial leaf which can be divided into sun-leaf and shade-leaf, there are obviously differences in structure between the two leave types.
2. There are great differences in the crude extraction rates of the leaves of if. corchorifolius in different months. The components, such as tannin, coumarin and flavone, vary in materials tested from different region. 49 compositions have been separated from its leaf essential oil; and 45 of them were identified. The content of (R) Butane. 1.2.4-trimethoxy reached 52.1%. Gallic acid was found in tannin, the kampfreol-3-O-β-D-6"-P- hydroxycoumaroyl-glucosie, one of flavonoid compounds, was firstly separated in the plant.
3. The leaf rude extraction in different months have different antibacterial activities to the 5 bacterium tested which in July is the best. Tannin is the best on inhibiting Shigella dusenterlea, the second on inhibiting Escherichia coli. Both are better than that of berberine. The antibacterial activity (except inhibiting Escherichia coli) of coumarin (50mg/mL) was better than that of berberine. Flavones have strong antibacterial activities, especially inhibiting Staphylococcus aureu. Essential oil has a strong sterilization effect; inhibiting Shigella dusenterlea is the best, inhibiting Escherichia coli the second. Sterilization effect of essential oil is a little better than that of berberine at the concentration of 0.5%.
4. There were no significant differences of liver index and spleen index between experimental group and control group, except the extract of tannin and coumarin of high dosage. The corrected phagocytic index (a value) of the flavone group was significantly higher than that of other groups (P≤5). It shows that the flavone in R. corchorifolius can enhance macrophage phagocytic activity. But there were no significant differences in phagocytic activity among each dosage of flavone. And there were no obvious differences in a value among the rest three groups and the CK. There were no pathological changes found by the observation of paraffin section of spleen, liver and kidney. It shows that the application of the compounds tested are safety to animals.
引文
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