僵蚕的生药学及药理活性研究
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摘要
中药的质量评价一直是中药研究与应用的难点与重点问题，制约着中药的现代化和国际化。僵蚕(Bombyx Batryticatus)是中医传统的平肝息风药。原名白僵蚕，亦称僵虫和天虫，来源于蚕蛾科(Bombycidae)昆虫家蚕(BombyxmoriL．)4～5龄的幼虫感染(或人工接种)白僵菌(Beauveria bassiana(Bals．)Vaill．)而致死的干燥全虫。僵蚕多于春、夏、秋三季生产，主要分布于四川、广西、江苏、浙江、安徽、山东、甘肃等省，且以四川省质量最优。僵蚕具有祛风定惊，化痰散结的功效。常用于惊风抽搐，咽喉肿痛，皮肤搔痒，颔下淋巴结炎，面神经麻痹等症。2005版中国药典对僵蚕的性状、粉末特征、性味与归经、功能与主治、用法与用量等方面进行了叙述。本文在此基础上对僵蚕的生药学及药理活性方面进行了较详细地研究，为僵蚕的研究增添了新的内容。
生药学方面，主要包括僵蚕的形成过程、采收加工、性状特征、显微特征、理化特征等方面。将四川省蚕业研究所提供的僵蚕作为标准品，与市售的10家药店的僵蚕进行了比较，并制定了以微量升华法鉴定药材僵蚕质量的新标准。
本文根据微量升华法的原理，自行设计了升华提取装置，可以大量提取到僵蚕升华物，经过理化分析、HPLC、IR、XRD等方法鉴定，此升华物为天然碳酸氢铵。升华物的急性毒性实验表明：分别采取灌胃和腹腔注射给药时，同样的给药剂量，灌胃时小鼠的最大耐受量为4．0／kg，但腹腔注射会使小鼠在短期内剧烈抽搐并且死亡，计算半数致死量为0．812g／kg。升华物的杀虫活性测定实验表明：熏蒸作用对蟋蟀和黄粉虫的效果明显。剂量越大，温度越高杀虫效果越好，但在超过试虫可适的高温条件下，正常组也有不同程度的死亡，说明温度过高对蟋蟀和黄粉虫也有很大影响。
抗惊厥药理实验表明：与正常组相比，僵蚕水提物(高剂量)可明显延长小鼠的惊厥潜伏时间(P<0．01)；僵蚕升华物(低剂量)可明显延长小鼠的死亡时间(P<0．01)；而白僵菌水提物(高剂量)明显缩短了小鼠的惊厥潜伏时间和死亡时间。说明僵蚕升华物为僵蚕的又一抗惊厥成分，同时也验证了白僵菌的毒性。
选取僵蚕升华物和石油醚层组分作为受试药对僵蚕的抗肿瘤活性进行了测定。实验表明：僵蚕升华物对肿瘤活性具有一定的抑制作用，低剂量和高剂量时的抑瘤率分别为43．56％和36．59％。但解剖时发现小鼠的脾脏发黑，说明升华物对H_(22)荷瘤小鼠的免疫器官有一定的负面影响。
体外抑菌实验主要选取了僵蚕升华物和石油醚层组分作为受试药进行了测定。实验表明：与正常组比较，僵蚕升华物对大肠杆菌ATCC25 923有非常显著的抑制作用(P<0．01)；僵蚕石油醚层组分对大肠杆菌ATCC25 923，大肠杆菌E_(cat)~R和所选取的金黄色葡萄球菌都有非常显著的抑制作用(P<0．01)。说明僵蚕升华物为僵蚕抑制大肠杆菌ATCC25 923的活性物质。僵蚕石油醚层组分中含有许多有效抑制大肠杆菌和金黄色葡萄球菌的活性成分，在今后的研究中可对此组分进行分离。
Quality evaluation of traditional Chinese medicine has been a difficult and multiple point on studying and applying for a long time,which has restricted the modernization and internationalization of traditional Chinese medicine.Bombyx Batryticatus is a traditional drug for calming the liver and subduing wind of Chinese medicine.Its original name is Jiang can.It can also be called Bai Jiangcan,Jiang Chong and Tian Chong.It is a dry and full body of 4-5 year old larva Bombyx mori L.which is origined from Bombycidae died of Beauveria bassiana (Bals.) Vaill.(or man-made).Bombyx Batryticatus is usually produced in spring,summer and autumn,which is mainly produced in Si Chuan province,Guang Xi province,Jiang Su province, Zhe Jiang province,An Hui province,Shan Dong province,Gan Su province and so on,and the quality of Si Chuan province is the best.Bombyx Batryticatus have effects such as dispelling wind,relieving convulsion,dissipate phlegm and eliminating stagnation.So,it is usually used for convulsion,Swelling of throat,skin titillation,lymphadenitis,facial paralysis and so on.It was described on characteristics,powder character,property and flavor meridian distribution, function and indication,usage and dosage in the Chinese Pharmacopeia of 2005.This paper had studied the pharmacognosy and pharmacological activity of Bombyx Batryticatus detailly, which increased new contents of Bombyx Batryticatus.
On the pharmacognosy of Bombyx Batryticatus,we mainly studied its formation, collection and processing,characteristics,micro-character,physico-character and chemicharacters. This paper used Bombyx Batryticatus supplied by institute of silkworm in Si Chuan province as standard,and compared it with Bombyx Batryticatus bought from 10 different drugstores in Chang Chun,and discovered a new quality standard on identifying Bombyx Batryticatus by microsublimation.
In this paper,according to the principle of microsublimation,we designed an new extracting device for sublimation.By the method,we could extract a great quantity of sublimation from Bombyx Batryticatus.The sublimation was identified as ammonium bicarbonate by the methods of high performance liquid chromatogram,infrared spectra and Xray diffraction.The acute toxicity test of the sublimation indicated that in the same dosage,the maximum tolerated dose of small mice was 4.0g/kg,when we used intragastric administration, and small mice twitched vigorously and died in a short time,when we used intraperitoneal injection.The median lethal dose was 0.812g/kg of intraperitoneal injection.The insecticidal activity test of the sublimation indicated that fumigating had an evident effect on Velar ifictorus rnicado(S.) and Tenebrio molitor(L.),The higher the does and temperature was,the better the insecticidal effect was.But in the condition of that insects could not accommodate,the control group had a different level of death.So megatemperature also had a large effect on Velarifictorus rnicado(S.) and Tenebrio molitor(L.).
In the anti-convulsion test,compared with the control group,the water extract of Bombyx Batryticatus(high dose) could prolong the convulsion latency of small mice obviously(P<0.01); the sublimation of Bombyx Batryticatus(low dose) could prolong the death time of small mice obviously(P<0.01);but the water extract of Beauveria bassiana.(Bals.) Vuilllt(high dose) decurtated the convulsion latency and death time of small mice obviously.It showed the toxicity of Beauveria bassiana.(Bals.) Vuilllt,and that the sublimation was also an anti-convulsion component of Bombyx Batryticatus.
The sublimation and petroleum ether fraction were examined to the anti-tumor activity of Bombyx Batryticatus.The result indicated that the sublimation of Bombyx Batryticatus had some inhibiting activity to tumor.The inhibition ratio of low dose was 43.56%and which of high dose was 36.59%.But we discovered that spleens of small mice became black.It showed that the sublimation had some negative effects on immune system of H_(22) small mice.
In the antibacterial activity test,the sublimation and petroleum ether fraction of Bombyx Batryticatus were examined.The result indicated that compared whith the control group,the sublimation had a very visible antibacterial activity to Escherichia coli ATCC25 923(P<0.01); The petroleum ether fraction had a very visible antibacterial activity(P<0.01) to Escherichia coli ATCC25 923,Escherichia coli E~R_(ca1) and Staphylococcus aureus we selected In conclusion,the sublimation is a active substance of Bombyx Batryticatus.And there were also some active components which could inhibit Escherichia coli and Staphylococcus aureus in the petroleum ether fraction of Bombyx Batryticatus.We should separate them in the future.
生药学方面，主要包括僵蚕的形成过程、采收加工、性状特征、显微特征、理化特征等方面。将四川省蚕业研究所提供的僵蚕作为标准品，与市售的10家药店的僵蚕进行了比较，并制定了以微量升华法鉴定药材僵蚕质量的新标准。
本文根据微量升华法的原理，自行设计了升华提取装置，可以大量提取到僵蚕升华物，经过理化分析、HPLC、IR、XRD等方法鉴定，此升华物为天然碳酸氢铵。升华物的急性毒性实验表明：分别采取灌胃和腹腔注射给药时，同样的给药剂量，灌胃时小鼠的最大耐受量为4．0／kg，但腹腔注射会使小鼠在短期内剧烈抽搐并且死亡，计算半数致死量为0．812g／kg。升华物的杀虫活性测定实验表明：熏蒸作用对蟋蟀和黄粉虫的效果明显。剂量越大，温度越高杀虫效果越好，但在超过试虫可适的高温条件下，正常组也有不同程度的死亡，说明温度过高对蟋蟀和黄粉虫也有很大影响。
抗惊厥药理实验表明：与正常组相比，僵蚕水提物(高剂量)可明显延长小鼠的惊厥潜伏时间(P<0．01)；僵蚕升华物(低剂量)可明显延长小鼠的死亡时间(P<0．01)；而白僵菌水提物(高剂量)明显缩短了小鼠的惊厥潜伏时间和死亡时间。说明僵蚕升华物为僵蚕的又一抗惊厥成分，同时也验证了白僵菌的毒性。
选取僵蚕升华物和石油醚层组分作为受试药对僵蚕的抗肿瘤活性进行了测定。实验表明：僵蚕升华物对肿瘤活性具有一定的抑制作用，低剂量和高剂量时的抑瘤率分别为43．56％和36．59％。但解剖时发现小鼠的脾脏发黑，说明升华物对H_(22)荷瘤小鼠的免疫器官有一定的负面影响。
体外抑菌实验主要选取了僵蚕升华物和石油醚层组分作为受试药进行了测定。实验表明：与正常组比较，僵蚕升华物对大肠杆菌ATCC25 923有非常显著的抑制作用(P<0．01)；僵蚕石油醚层组分对大肠杆菌ATCC25 923，大肠杆菌E_(cat)~R和所选取的金黄色葡萄球菌都有非常显著的抑制作用(P<0．01)。说明僵蚕升华物为僵蚕抑制大肠杆菌ATCC25 923的活性物质。僵蚕石油醚层组分中含有许多有效抑制大肠杆菌和金黄色葡萄球菌的活性成分，在今后的研究中可对此组分进行分离。
Quality evaluation of traditional Chinese medicine has been a difficult and multiple point on studying and applying for a long time,which has restricted the modernization and internationalization of traditional Chinese medicine.Bombyx Batryticatus is a traditional drug for calming the liver and subduing wind of Chinese medicine.Its original name is Jiang can.It can also be called Bai Jiangcan,Jiang Chong and Tian Chong.It is a dry and full body of 4-5 year old larva Bombyx mori L.which is origined from Bombycidae died of Beauveria bassiana (Bals.) Vaill.(or man-made).Bombyx Batryticatus is usually produced in spring,summer and autumn,which is mainly produced in Si Chuan province,Guang Xi province,Jiang Su province, Zhe Jiang province,An Hui province,Shan Dong province,Gan Su province and so on,and the quality of Si Chuan province is the best.Bombyx Batryticatus have effects such as dispelling wind,relieving convulsion,dissipate phlegm and eliminating stagnation.So,it is usually used for convulsion,Swelling of throat,skin titillation,lymphadenitis,facial paralysis and so on.It was described on characteristics,powder character,property and flavor meridian distribution, function and indication,usage and dosage in the Chinese Pharmacopeia of 2005.This paper had studied the pharmacognosy and pharmacological activity of Bombyx Batryticatus detailly, which increased new contents of Bombyx Batryticatus.
On the pharmacognosy of Bombyx Batryticatus,we mainly studied its formation, collection and processing,characteristics,micro-character,physico-character and chemicharacters. This paper used Bombyx Batryticatus supplied by institute of silkworm in Si Chuan province as standard,and compared it with Bombyx Batryticatus bought from 10 different drugstores in Chang Chun,and discovered a new quality standard on identifying Bombyx Batryticatus by microsublimation.
In this paper,according to the principle of microsublimation,we designed an new extracting device for sublimation.By the method,we could extract a great quantity of sublimation from Bombyx Batryticatus.The sublimation was identified as ammonium bicarbonate by the methods of high performance liquid chromatogram,infrared spectra and Xray diffraction.The acute toxicity test of the sublimation indicated that in the same dosage,the maximum tolerated dose of small mice was 4.0g/kg,when we used intragastric administration, and small mice twitched vigorously and died in a short time,when we used intraperitoneal injection.The median lethal dose was 0.812g/kg of intraperitoneal injection.The insecticidal activity test of the sublimation indicated that fumigating had an evident effect on Velar ifictorus rnicado(S.) and Tenebrio molitor(L.),The higher the does and temperature was,the better the insecticidal effect was.But in the condition of that insects could not accommodate,the control group had a different level of death.So megatemperature also had a large effect on Velarifictorus rnicado(S.) and Tenebrio molitor(L.).
In the anti-convulsion test,compared with the control group,the water extract of Bombyx Batryticatus(high dose) could prolong the convulsion latency of small mice obviously(P<0.01); the sublimation of Bombyx Batryticatus(low dose) could prolong the death time of small mice obviously(P<0.01);but the water extract of Beauveria bassiana.(Bals.) Vuilllt(high dose) decurtated the convulsion latency and death time of small mice obviously.It showed the toxicity of Beauveria bassiana.(Bals.) Vuilllt,and that the sublimation was also an anti-convulsion component of Bombyx Batryticatus.
The sublimation and petroleum ether fraction were examined to the anti-tumor activity of Bombyx Batryticatus.The result indicated that the sublimation of Bombyx Batryticatus had some inhibiting activity to tumor.The inhibition ratio of low dose was 43.56%and which of high dose was 36.59%.But we discovered that spleens of small mice became black.It showed that the sublimation had some negative effects on immune system of H_(22) small mice.
In the antibacterial activity test,the sublimation and petroleum ether fraction of Bombyx Batryticatus were examined.The result indicated that compared whith the control group,the sublimation had a very visible antibacterial activity to Escherichia coli ATCC25 923(P<0.01); The petroleum ether fraction had a very visible antibacterial activity(P<0.01) to Escherichia coli ATCC25 923,Escherichia coli E~R_(ca1) and Staphylococcus aureus we selected In conclusion,the sublimation is a active substance of Bombyx Batryticatus.And there were also some active components which could inhibit Escherichia coli and Staphylococcus aureus in the petroleum ether fraction of Bombyx Batryticatus.We should separate them in the future.
引文
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