灰蔸巴多糖的提取及其降血糖活性的研究
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摘要
灰蔸巴是一种寄生在海拔1 500米以上的峨眉山茶树干上的红蜘蛛吐丝筑的巢(也有称其为老茶树头部生长出来的菌科植物),亦称其为灰兜巴、灰蔸芭、闭口袋,在四川峨眉山地区用此药水煎液治疗糖尿病,效果较好。关于该药的疗效、有效成分及药理研究,并未有详尽的研究报道。
本文对灰蔸巴提取液进行了药效研究,确定其降血糖的有效成分为灰蔸巴多糖,确定灰蔸巴的降血糖疗效,同时对有效降血糖成分进行分离提取工艺研究以及纯化。为临床糖尿病的药物治疗提供新的线索,对开发和研究治疗糖尿病的新物质,特别是从天然资源中找到既能治疗糖尿病又无明显不良反应的药物,具有特别重要的意义。
在进行灰蔸巴降血糖研究中以四氧嘧啶诱导糖尿病动物模型,建立了稳定可靠的动物模型。在检测血糖指标的同时,检测了实验动物血清及脏器中的SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN等相关指标,服用灰蔸巴的糖尿病模型动物其血糖值下降,血脂接近正常,抗氧化能力有所提高,说明灰蔸巴多糖不仅具有降血糖疗效,同时可提高机体的抗氧化活性,增加机体降血脂能力,灰蔸巴能改善受损胰岛β细胞的功能。
本文首次采用先给予健康实验动物灌胃灰蔸巴多糖,再用四氧嘧啶诱导糖尿病模型的方式,确定灰蔸巴多糖具有预防由四氧嘧啶引起的糖尿病。通过其血清及脏器中的SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN等相关指标,可以得看出服用灰蔸巴多糖的实验动物其抗氧化和降脂能力均有所提高,说明灰蔸巴多糖能够保护胰岛β细胞免遭四氧嘧啶的损害。
本文对灰蔸巴进行了大量的毒理实验,说明灰蔸巴是一种药性缓和、毒副作用小的中药材,具有广阔的应用前景。
Huidouba, which is an effective Chinese traditional medicine to treat the diabete in local, is the cobweb that secreted by parasitic spider on the tree at an elevation of 1500m on Emei mountain in Sichuan province. The extraction process of Huidouba polysaccharide and its anti-diabetic activity were studied in this work. There is no report on this field until now.
The method to induce the diabetic animals was investigated at first. After the animals were feed for 3 days and drunk sugar water, the freshly prepared solutions of Alloxan (140mg/kg body weight) wereintraperitoneally injected to the 16 hours fasted mice, while 120mg/kg first day and 100mg/kg next day for rats. Three hours after that, the animals were fed with 25% glucose solution(1ml/100g), then, the blood glucose levels were tested with blood sample from orbital venous plexus. The animal was selected as diabetic model whose glucose level was higher than 11.1mM and sustained at that level. In the period of feeding, fresh water and clean living conditions must be guaranteed. paddings were changed every two days and water was supplied every day. Animals were kept in a special animal house at an ambient temperature of 22±1℃and 50-65% relative humidity. The induction rate was connected with the amount of Alloxan, injection way and feeding method.
The chemical identification results indecate that Huidouba consists polysaccharide and no protein, Huidouba may contain steroid, phenols, tannins, saponins and alkaloids. So, we extracted crude polysaccharide in traditional method. In our research, the Huidouba polysaccharide (HDBW-1) could decrease the blood glucose level of alloxan-induced diabetic mice by 44.3% and decrease the blood glucose level of alloxan-induced diabetic rats by 42.4%. Significantly, the hypoglycemic effect of Huidouba polysaccharide(HDBW-1) is superior to that of Huidouba ethanol extract(HDBE) and Huidouba water extract(HDBW).
Moreover, the Huidouba polysaccharide exhibits no obvious hypoglycemic effects on normal mice or rats(P>0.05). So we could draw the conclusion that the effective component of Huidouba is polysaccharide(HDBW-1).
Huidouba polysaccharide was extracted in this work. The extraction process was optimizated by response surface method. I studied the effect of Huidouba polysaccharide on diabetic animals. The results show that Huidouba polysacharide has obvious hypocemic effects on diabetic animals. Furthermore, It is shown that the anti-oxidation and lipid lowing activities increased from the changes of SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN after treatment with Huidouba polysaccharide. The healthy animals were fed by Huidouba polysaccharide for 30 days, then, the animals was induced to diabete by Alloxan. There is no significant difference between the blood glucose levels before and after induction. It is shown that the Huidouba polysaccharide could prevent the diabete. In another experiment, the results showed that the Huidouba polysaccharide could treat the diabete because the glucose level of diabetic animals decreased after they were fed with Huidouba polysaccharide. It is shown that the anti-oxidation and lipid lowing activities increased from the changes of SOD、GSH-Px、MDA、TG、TCHO、LDL-C、HDL-C、BUN after treatment with Huidouba polysaccharide. It could be concluded that the Huidouba polysaccharide could protect isletβcell from the injury of Alloxan.
Through isolation and purification, the Huidouba polysaccharide was devided into three components-HDBW-1Cα、HDBW-1Cβand HDBW-1Cγ.The monosaccharide composition of all the three components are the same-mannose, rhamnse, glucose, galactose, xylose, arabinose, fucose, glucuronic acid and galacturonic acid.
At last, it is also shown there is little toxicity for Huidouba polysaccharide after the acute, sub-acute, long term toxcicity experiments and special toxcicity experiments. It would be a potential drug to treat the diabetes.
本文对灰蔸巴提取液进行了药效研究,确定其降血糖的有效成分为灰蔸巴多糖,确定灰蔸巴的降血糖疗效,同时对有效降血糖成分进行分离提取工艺研究以及纯化。为临床糖尿病的药物治疗提供新的线索,对开发和研究治疗糖尿病的新物质,特别是从天然资源中找到既能治疗糖尿病又无明显不良反应的药物,具有特别重要的意义。
在进行灰蔸巴降血糖研究中以四氧嘧啶诱导糖尿病动物模型,建立了稳定可靠的动物模型。在检测血糖指标的同时,检测了实验动物血清及脏器中的SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN等相关指标,服用灰蔸巴的糖尿病模型动物其血糖值下降,血脂接近正常,抗氧化能力有所提高,说明灰蔸巴多糖不仅具有降血糖疗效,同时可提高机体的抗氧化活性,增加机体降血脂能力,灰蔸巴能改善受损胰岛β细胞的功能。
本文首次采用先给予健康实验动物灌胃灰蔸巴多糖,再用四氧嘧啶诱导糖尿病模型的方式,确定灰蔸巴多糖具有预防由四氧嘧啶引起的糖尿病。通过其血清及脏器中的SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN等相关指标,可以得看出服用灰蔸巴多糖的实验动物其抗氧化和降脂能力均有所提高,说明灰蔸巴多糖能够保护胰岛β细胞免遭四氧嘧啶的损害。
本文对灰蔸巴进行了大量的毒理实验,说明灰蔸巴是一种药性缓和、毒副作用小的中药材,具有广阔的应用前景。
Huidouba, which is an effective Chinese traditional medicine to treat the diabete in local, is the cobweb that secreted by parasitic spider on the tree at an elevation of 1500m on Emei mountain in Sichuan province. The extraction process of Huidouba polysaccharide and its anti-diabetic activity were studied in this work. There is no report on this field until now.
The method to induce the diabetic animals was investigated at first. After the animals were feed for 3 days and drunk sugar water, the freshly prepared solutions of Alloxan (140mg/kg body weight) wereintraperitoneally injected to the 16 hours fasted mice, while 120mg/kg first day and 100mg/kg next day for rats. Three hours after that, the animals were fed with 25% glucose solution(1ml/100g), then, the blood glucose levels were tested with blood sample from orbital venous plexus. The animal was selected as diabetic model whose glucose level was higher than 11.1mM and sustained at that level. In the period of feeding, fresh water and clean living conditions must be guaranteed. paddings were changed every two days and water was supplied every day. Animals were kept in a special animal house at an ambient temperature of 22±1℃and 50-65% relative humidity. The induction rate was connected with the amount of Alloxan, injection way and feeding method.
The chemical identification results indecate that Huidouba consists polysaccharide and no protein, Huidouba may contain steroid, phenols, tannins, saponins and alkaloids. So, we extracted crude polysaccharide in traditional method. In our research, the Huidouba polysaccharide (HDBW-1) could decrease the blood glucose level of alloxan-induced diabetic mice by 44.3% and decrease the blood glucose level of alloxan-induced diabetic rats by 42.4%. Significantly, the hypoglycemic effect of Huidouba polysaccharide(HDBW-1) is superior to that of Huidouba ethanol extract(HDBE) and Huidouba water extract(HDBW).
Moreover, the Huidouba polysaccharide exhibits no obvious hypoglycemic effects on normal mice or rats(P>0.05). So we could draw the conclusion that the effective component of Huidouba is polysaccharide(HDBW-1).
Huidouba polysaccharide was extracted in this work. The extraction process was optimizated by response surface method. I studied the effect of Huidouba polysaccharide on diabetic animals. The results show that Huidouba polysacharide has obvious hypocemic effects on diabetic animals. Furthermore, It is shown that the anti-oxidation and lipid lowing activities increased from the changes of SOD、GSH-Px、MDA、TG、TC、LDL-C、HDL-C、BUN after treatment with Huidouba polysaccharide. The healthy animals were fed by Huidouba polysaccharide for 30 days, then, the animals was induced to diabete by Alloxan. There is no significant difference between the blood glucose levels before and after induction. It is shown that the Huidouba polysaccharide could prevent the diabete. In another experiment, the results showed that the Huidouba polysaccharide could treat the diabete because the glucose level of diabetic animals decreased after they were fed with Huidouba polysaccharide. It is shown that the anti-oxidation and lipid lowing activities increased from the changes of SOD、GSH-Px、MDA、TG、TCHO、LDL-C、HDL-C、BUN after treatment with Huidouba polysaccharide. It could be concluded that the Huidouba polysaccharide could protect isletβcell from the injury of Alloxan.
Through isolation and purification, the Huidouba polysaccharide was devided into three components-HDBW-1Cα、HDBW-1Cβand HDBW-1Cγ.The monosaccharide composition of all the three components are the same-mannose, rhamnse, glucose, galactose, xylose, arabinose, fucose, glucuronic acid and galacturonic acid.
At last, it is also shown there is little toxicity for Huidouba polysaccharide after the acute, sub-acute, long term toxcicity experiments and special toxcicity experiments. It would be a potential drug to treat the diabetes.
引文
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