蜜蜂巢脾挥发油及其治疗AR的药效学研究
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摘要
蜜蜂巢脾是我国传统的动物中药。具有多种生物学功能和药理学价值。然而，从目前来看，对巢脾有效成分及其药理的研究还是经验多，科学验证少，定性定量少，蜜蜂蜂巢具有治疗鼻炎特别是变应性鼻炎(AR)的作用，然而药效成分和作用机理还不是十分清楚。关于蜜蜂巢脾挥发油的成分研究、药理作用研究国内外尚未报道过。本文对蜜蜂巢脾挥发油进行定性定量分析，对其提取工艺、提取动力学进行优选和数学模拟，并对其治疗AR药效学进行研究。结果如下：
1对蜜蜂巢脾挥发油提取工艺进行了优选研究，得出较佳提取工艺为蜜蜂巢脾粉碎度为Ⅱ、用8倍量水，浸泡20h、提取12h。建立并确定提取动力学方程ln(⊿A_t／⊿_t)=-0.1904t_c-2.9325(R=0.9936)和ln(0.2768-A_t)=-0.1947t-1.2934(R=0.9997)。为蜜蜂巢脾挥发油开发利用提供基础参数数据和有益参考。
2采用气相色谱、质谱联用(GC／MG)技术，首次对意大利蜜蜂巢脾挥发油组成成分进行了分析鉴定，分离鉴定出46种化合物。为蜜蜂巢脾有效成分明确化奠定基础。
3蜜蜂巢脾醇提物(15kg／kg)和蜜蜂巢脾挥发油(1mL／kg、2mL／kg、3mL／kg)均能够显著抑制小鼠二甲苯所致耳肿胀(p＜0.05)、抑制小鼠醋酸所致腹腔毛细血管通透性(p＜0.05)，并呈良好的量效关系。揭示蜜蜂巢脾挥发油能够抑制炎性水肿、抑制渗出和血管通透性增加的急性炎症，具有抗炎作用。
4蜜蜂巢脾醇提物(15kg／kg)和蜜蜂巢脾挥发油(1mL／kg、2mL／kg、3mL／kg)均能够提高小鼠脾脏指数、胸腺指数，且呈良好的量效关系，揭示蜜蜂巢脾挥发油能够通过免疫器官发挥免疫增强作用。是其免疫增强作用机制之一。
5蜜蜂巢脾醇提物及挥发油对实验性AR有确切治疗作用：能缓减AR局部症状，改善局部粘膜水肿，减少肥大细胞和嗜酸性细胞在炎症局部的浸润、抑制AR模型大鼠血清炎性介质组胺的释放。免疫法测定血清中总IgA、总IgM和总IgG含量，表现为蜜蜂巢脾醇提物及挥发油能够显著升高血清IgA、IgG水平，揭示封闭性抗体可与进入机体的抗原物质优先结合，竞争抑制IgE抗体，调节粘膜免疫，从而对AR具有治疗作用的可能机制。
6蜜蜂巢脾挥发油成分明确，蜜蜂巢脾醇提物及挥发油具有抗炎免疫作用，对AR疗效确切，有望成为治疗此病的新型药物。
Honeybee comb ,one kind of Chinese traditional animal medicine, is valuable for its biological function and pharmacology effect. However, the study on the functional compone nts of Honeybee comb and its pharmacology effect are more experiential, less scientific validations, and less qualitative and quantitative analysis. Though honeybee comb has the effect on Allergic Rhinitis (AR), the functional components and its mechanisms are not clear. The reports about the volatile oil which is the component of honeybee comb and the effect of the volatile oil on AR are not funded all over the world. The dissertation firstly studied the components of honeybee comb volatile oil .including the qualitative and quantitative analysis, chosed a best extraction condition of honeybee comb volatile oil , made mathematics kinetics models of the extraction,and studied the effect of honeybee comb volatile oil on the AR and the its parts of immunological mechanisms. The results of the experiment showed that:
1 A best extraction condition of honeybee comb volatile oil by vapor distillation areⅡcomminution degree, 8 times water of honeybee comb, dipping for 20h, distilling for 12h; The optimizing mat hematic kinetics model for the relationship between the yield and the extraction time during the extraction process of volatile oil from honeycomb are ln(0．2768--A_t) =-0．1947t-1．2934 (R= 0．9997) and ln((?)A_t/(?)t) =-0．1904t-2．9325 (R= 0．9936) . the kinetics model is useful for controlling the process of the extraction of the volatile oil from honeycomb.
2 Used the the combination technologis of gas chromatography and mass spectrum (GC/MS), 46 compounds were identified.
3 Both the ethanolic extracts from honeybee comb (15kg/kg) and the honeybee comb volatile oil (1mL/kg^ 2mL/kgx 3mL/kg)can restrain the mice ear edema caused by dimethylbenzene and the mice capillary permeability caused by acetic acid, the honeybee comb volatile oil has the good relationship between the dosages and the effect.the results showed that both of them have the effect of antiinflammation by controlling the acute inflammation which concluding the edema ,the capillary permeability and so on.
4 Both the ethanolic extracts from honeybee comb (15kg/kg) and the honeybee comb volatile oil (1mL/kg、2mL/kg、3mL/kg)can increase the index of mice' s thymus gland and the index of mice's spleen (p<0．05) .the honeybee comb volatile oil has the good relationship between the dosages and the effect, the results showed that strengthening the immunity apparatuses such as thymus gland and spleen may be one of the mechanisms of the compounds and the honeybee comb volatile oil increasing the capacity of immunity.
5 The therapy effect of Both the ethanolic extracts from honeybee comb and the honeybee comb volatile oil on AR was significant and clear : improving the symptoms of rats suffered from AR ; reducing the number of mast cell,the number of eosinophil and other inflammation cells ; reducing the level of rat's histamine of blood serum ; Both the ethanolic extracts from honeybee comb and the honeybee comb volatile oil can increase the levels of IgA and IgG of blood serum , It can be one of the immunology mechanisms of the effect on AR that IgA and IgG may combine with the antigen first in body, complete with IgE,restrain IgE action.and regulate the immunity of nasal mucosal.
6 In conclusion, the compounds of honeybee comb volatile oil is clear.The ethanolic extracts and the volatile oil from honeybee comb , have the effect of antiinflammation, can increase the capacity of immunity, both of which effect on AR is valid, and will be new medicines to AR.
1对蜜蜂巢脾挥发油提取工艺进行了优选研究，得出较佳提取工艺为蜜蜂巢脾粉碎度为Ⅱ、用8倍量水，浸泡20h、提取12h。建立并确定提取动力学方程ln(⊿A_t／⊿_t)=-0.1904t_c-2.9325(R=0.9936)和ln(0.2768-A_t)=-0.1947t-1.2934(R=0.9997)。为蜜蜂巢脾挥发油开发利用提供基础参数数据和有益参考。
2采用气相色谱、质谱联用(GC／MG)技术，首次对意大利蜜蜂巢脾挥发油组成成分进行了分析鉴定，分离鉴定出46种化合物。为蜜蜂巢脾有效成分明确化奠定基础。
3蜜蜂巢脾醇提物(15kg／kg)和蜜蜂巢脾挥发油(1mL／kg、2mL／kg、3mL／kg)均能够显著抑制小鼠二甲苯所致耳肿胀(p＜0.05)、抑制小鼠醋酸所致腹腔毛细血管通透性(p＜0.05)，并呈良好的量效关系。揭示蜜蜂巢脾挥发油能够抑制炎性水肿、抑制渗出和血管通透性增加的急性炎症，具有抗炎作用。
4蜜蜂巢脾醇提物(15kg／kg)和蜜蜂巢脾挥发油(1mL／kg、2mL／kg、3mL／kg)均能够提高小鼠脾脏指数、胸腺指数，且呈良好的量效关系，揭示蜜蜂巢脾挥发油能够通过免疫器官发挥免疫增强作用。是其免疫增强作用机制之一。
5蜜蜂巢脾醇提物及挥发油对实验性AR有确切治疗作用：能缓减AR局部症状，改善局部粘膜水肿，减少肥大细胞和嗜酸性细胞在炎症局部的浸润、抑制AR模型大鼠血清炎性介质组胺的释放。免疫法测定血清中总IgA、总IgM和总IgG含量，表现为蜜蜂巢脾醇提物及挥发油能够显著升高血清IgA、IgG水平，揭示封闭性抗体可与进入机体的抗原物质优先结合，竞争抑制IgE抗体，调节粘膜免疫，从而对AR具有治疗作用的可能机制。
6蜜蜂巢脾挥发油成分明确，蜜蜂巢脾醇提物及挥发油具有抗炎免疫作用，对AR疗效确切，有望成为治疗此病的新型药物。
Honeybee comb ,one kind of Chinese traditional animal medicine, is valuable for its biological function and pharmacology effect. However, the study on the functional compone nts of Honeybee comb and its pharmacology effect are more experiential, less scientific validations, and less qualitative and quantitative analysis. Though honeybee comb has the effect on Allergic Rhinitis (AR), the functional components and its mechanisms are not clear. The reports about the volatile oil which is the component of honeybee comb and the effect of the volatile oil on AR are not funded all over the world. The dissertation firstly studied the components of honeybee comb volatile oil .including the qualitative and quantitative analysis, chosed a best extraction condition of honeybee comb volatile oil , made mathematics kinetics models of the extraction,and studied the effect of honeybee comb volatile oil on the AR and the its parts of immunological mechanisms. The results of the experiment showed that:
1 A best extraction condition of honeybee comb volatile oil by vapor distillation areⅡcomminution degree, 8 times water of honeybee comb, dipping for 20h, distilling for 12h; The optimizing mat hematic kinetics model for the relationship between the yield and the extraction time during the extraction process of volatile oil from honeycomb are ln(0．2768--A_t) =-0．1947t-1．2934 (R= 0．9997) and ln((?)A_t/(?)t) =-0．1904t-2．9325 (R= 0．9936) . the kinetics model is useful for controlling the process of the extraction of the volatile oil from honeycomb.
2 Used the the combination technologis of gas chromatography and mass spectrum (GC/MS), 46 compounds were identified.
3 Both the ethanolic extracts from honeybee comb (15kg/kg) and the honeybee comb volatile oil (1mL/kg^ 2mL/kgx 3mL/kg)can restrain the mice ear edema caused by dimethylbenzene and the mice capillary permeability caused by acetic acid, the honeybee comb volatile oil has the good relationship between the dosages and the effect.the results showed that both of them have the effect of antiinflammation by controlling the acute inflammation which concluding the edema ,the capillary permeability and so on.
4 Both the ethanolic extracts from honeybee comb (15kg/kg) and the honeybee comb volatile oil (1mL/kg、2mL/kg、3mL/kg)can increase the index of mice' s thymus gland and the index of mice's spleen (p<0．05) .the honeybee comb volatile oil has the good relationship between the dosages and the effect, the results showed that strengthening the immunity apparatuses such as thymus gland and spleen may be one of the mechanisms of the compounds and the honeybee comb volatile oil increasing the capacity of immunity.
5 The therapy effect of Both the ethanolic extracts from honeybee comb and the honeybee comb volatile oil on AR was significant and clear : improving the symptoms of rats suffered from AR ; reducing the number of mast cell,the number of eosinophil and other inflammation cells ; reducing the level of rat's histamine of blood serum ; Both the ethanolic extracts from honeybee comb and the honeybee comb volatile oil can increase the levels of IgA and IgG of blood serum , It can be one of the immunology mechanisms of the effect on AR that IgA and IgG may combine with the antigen first in body, complete with IgE,restrain IgE action.and regulate the immunity of nasal mucosal.
6 In conclusion, the compounds of honeybee comb volatile oil is clear.The ethanolic extracts and the volatile oil from honeybee comb , have the effect of antiinflammation, can increase the capacity of immunity, both of which effect on AR is valid, and will be new medicines to AR.
引文
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