泰和乌骨鸡多肽若干功能活性研究
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摘要
泰和乌骨鸡是我国传统公认的天然药用鸡种,主要产于江西省泰和县。作为传统中药,乌骨鸡既可以单独食用也可以配药入复方,具有补肝肾、益气血、退虚热、治心腹痛等功效。目前,对于乌骨鸡鸡肉酶解获得多肽的功能活性研究还少有报道。此外,作为中国传统中药,乌骨鸡多肽是否与其药用机理有关值得进行全面的实验研究。鉴于此,本文建立多种小鼠模型,对乌骨鸡多肽的补血、抗炎、镇痛及肝脏保护功能进行了研究。现将本文主要研究内容与结果概括如下：
1.建立了失血法和氟尿嘧啶注射法联合致血虚小鼠模型,对泰和乌骨鸡多肽的补血活性进行了研究,以评价乌骨鸡多肽及其一种分离多肽对血虚小鼠的补血作用。研究测定了实验小鼠红细胞和血红蛋白水平。结果显示,活性肽组RBC水平在第12天时与血虚组相比差异显著(P<0.05)；活性肽组在第6至12天期间升高RBC和HGB水平作用与阿胶组相比明显加快,具有显著性差异(P<0.05)。第18天时,活性肽组HGB水平已超过各组水平,且与正常组相比具有显著性差异(P<0.05)。活性肽组分A组的RBC水平在各个时间点均高于血虚组,但均无显著性差异(P>0.05)。活性肽组分A组在第20天时HGB水平已与正常组无显著性差异(P>0.05),而血虚组和阿胶组仍与正常组有极显著差异(P<0.01)。
2.建立了小鼠足跖肿胀模型、耳二甲苯致炎模型、毛细血管通透性模型,对泰和乌骨鸡多肽的抗炎活性进行了研究。结果发现,乌骨鸡多肽能够显著性(P<0.05)地抑制机体PGE2的合成或释放,但是对于机体的表观肿胀现象抑制效果不佳(P>0.05),表明乌骨鸡多肽具有一定的抗炎功能活性。
3.建立了小鼠热板法实验模型和醋酸扭体法实验模型,对泰和乌骨鸡多肽的镇痛功能活性进行了研究。结果发现,虽然乌骨鸡多肽仅表现出较弱的作用效果,没有显著提高扭体潜伏期,但低剂量和高剂量的乌骨鸡多肽都显著(P<0.01)抑制了小鼠扭体反应次数,且扭体抑制率都达到了68.4%,表明乌骨鸡多肽具有一定抑制化学性损伤的镇痛作用。对于热板法模型,乌骨鸡多肽的镇痛作用不明显(P>0.05),仅有较低的镇痛趋势。表明乌骨鸡多肽对于化学性损伤所致疼痛的抑制作用效果较好,而对热痛刺激的抑制效果则不及前者。
4.建立了小鼠四氯化碳肝损伤模型和D-半乳糖胺肝损伤模型,对泰和乌骨鸡多肽的肝脏保护功能活性进行了研究。结果发现,乌骨鸡多肽对两种模型小鼠的ALT、AST、MDA、SOD、肝指数、脾指数指标作用效果不显著(P>0.05),对小鼠肝损伤没有起到显著的保护作用,对机体脂质过氧化的抵抗作用不明显,对模型药物导致的小鼠肝、脾增大没有显著的抑制作用(P>0.05)。
Taihe black-bone silky fowl (TBSF) is an acknowledged breed of natural medical chicken in Chinese tradition, grows primarily in South China, Taihe, Jiangxi Province. As a traditional Chinese medicine, it can be used independently or in compound prescription, BSF has been proved to have the effects of nourishing liver and kidney, profiting vitality, reducing asthenia heat, curing abdominal pain and so on. At present, the bioactive research of the peptides, which is acquired from the meat of TBSF by enzymolysis, is still reported infrequently. It is worth of an all-sided experiment research, to find out whether there is any relationship between the TBSF peptides and the medicinal mechanism of TBSF. In consideration of aforementioned, this paper established several animal modes to study on the hemopoietic effect, anti-inflammatory effect, analgesic effect and liver protective effect of TBSF peptides. Now the main research content is summarized as follows:
1. To observe the hematopoiesis activity of the TBSF peptide and one of its fractions (A), blood loss combined with myelosuppression drug (Fluorouracil,5-FU) were used to construct the blood deficiency mice model, and the red blood cells (RBC) and hemoglobin (HGB) levels were measured. Results showed that, on the 12th day, the RBC level of the bioactive peptides group has a significant difference (P<0.05) compared with the blood deficiency group; During the period of 6 to 12 days, the effect of rising the RBC and HGB level of the bioactive peptides group is significantly quicker than the Ejiao group (P<0.05). On the 18th day, the HGB level of the bioactive peptides group exceeds the other groups, and is significantly different (P<0.05) from the normal control group. The RBC level of the bioactive peptides component A group is higher than the blood deficiency group, without significant difference (P>0.05), while the blood deficiency group and the Ejiao group are significantly different (P<0.01) from the normal control group. On the 20th day, there is no significant difference (P>0.05) between the bioactive peptides component A group and the normal control group on the level of HGB, while the blood deficiency group and the Ejiao group are significantly different (P<0.01) from the normal control group.
2. To observe the anti-inflammatory activity of the TBSF peptide, the carrageenan-induced paw edema in mice model, ear xylene cause inflammation model and capillary permeability model were used in the research. The result showed that, the peptides of TBSF could inhibit the synthesis or releases of PGE2 in body tissues significantly, but to the apparent edema of body, no significant inhibit effect was found. Those suggest that the peptides of TBSF exhibited anti-inflammatory activities to some extent.
3. Investigations on the analgesic effects were conducted, including hot-plate test model and acetic acid-induced writhing test model. The results showed that the low and high doses of TBSF peptides did not increase the latency time of mice significantly, only a weak effect of analgesic was showed, but the peptides could significantly decreased the number of writhing, and inhibited the writhing response by 68.4%, showed their analgesic effects to chemical injury. To the hot-plate test model, the analgesic effect was not so significant, a low analgesic tendency exhibited. Those suggest that TBSF peptide has a better analgesic effect to chemical injury, rather than the thermal pain stimulus.
4. To study on the liver protective effect of TBSF peptides, CCI4 and D-galactosamine induced hepatic injury model were established respectively. The results were found that the ALT, AST, MDA, LI, and SI indexes were not significantly inhibited, and the SOD level was not increased. Those means remarkable protective effect was not showed to the liver injury, the effect on the inhibition of lipid peroxidation to body tissues was inconspicuous, and peptides did not show a prominent inhibitory action to the pharmaceuticals induced liver and spleen accretion.
1.建立了失血法和氟尿嘧啶注射法联合致血虚小鼠模型,对泰和乌骨鸡多肽的补血活性进行了研究,以评价乌骨鸡多肽及其一种分离多肽对血虚小鼠的补血作用。研究测定了实验小鼠红细胞和血红蛋白水平。结果显示,活性肽组RBC水平在第12天时与血虚组相比差异显著(P<0.05)；活性肽组在第6至12天期间升高RBC和HGB水平作用与阿胶组相比明显加快,具有显著性差异(P<0.05)。第18天时,活性肽组HGB水平已超过各组水平,且与正常组相比具有显著性差异(P<0.05)。活性肽组分A组的RBC水平在各个时间点均高于血虚组,但均无显著性差异(P>0.05)。活性肽组分A组在第20天时HGB水平已与正常组无显著性差异(P>0.05),而血虚组和阿胶组仍与正常组有极显著差异(P<0.01)。
2.建立了小鼠足跖肿胀模型、耳二甲苯致炎模型、毛细血管通透性模型,对泰和乌骨鸡多肽的抗炎活性进行了研究。结果发现,乌骨鸡多肽能够显著性(P<0.05)地抑制机体PGE2的合成或释放,但是对于机体的表观肿胀现象抑制效果不佳(P>0.05),表明乌骨鸡多肽具有一定的抗炎功能活性。
3.建立了小鼠热板法实验模型和醋酸扭体法实验模型,对泰和乌骨鸡多肽的镇痛功能活性进行了研究。结果发现,虽然乌骨鸡多肽仅表现出较弱的作用效果,没有显著提高扭体潜伏期,但低剂量和高剂量的乌骨鸡多肽都显著(P<0.01)抑制了小鼠扭体反应次数,且扭体抑制率都达到了68.4%,表明乌骨鸡多肽具有一定抑制化学性损伤的镇痛作用。对于热板法模型,乌骨鸡多肽的镇痛作用不明显(P>0.05),仅有较低的镇痛趋势。表明乌骨鸡多肽对于化学性损伤所致疼痛的抑制作用效果较好,而对热痛刺激的抑制效果则不及前者。
4.建立了小鼠四氯化碳肝损伤模型和D-半乳糖胺肝损伤模型,对泰和乌骨鸡多肽的肝脏保护功能活性进行了研究。结果发现,乌骨鸡多肽对两种模型小鼠的ALT、AST、MDA、SOD、肝指数、脾指数指标作用效果不显著(P>0.05),对小鼠肝损伤没有起到显著的保护作用,对机体脂质过氧化的抵抗作用不明显,对模型药物导致的小鼠肝、脾增大没有显著的抑制作用(P>0.05)。
Taihe black-bone silky fowl (TBSF) is an acknowledged breed of natural medical chicken in Chinese tradition, grows primarily in South China, Taihe, Jiangxi Province. As a traditional Chinese medicine, it can be used independently or in compound prescription, BSF has been proved to have the effects of nourishing liver and kidney, profiting vitality, reducing asthenia heat, curing abdominal pain and so on. At present, the bioactive research of the peptides, which is acquired from the meat of TBSF by enzymolysis, is still reported infrequently. It is worth of an all-sided experiment research, to find out whether there is any relationship between the TBSF peptides and the medicinal mechanism of TBSF. In consideration of aforementioned, this paper established several animal modes to study on the hemopoietic effect, anti-inflammatory effect, analgesic effect and liver protective effect of TBSF peptides. Now the main research content is summarized as follows:
1. To observe the hematopoiesis activity of the TBSF peptide and one of its fractions (A), blood loss combined with myelosuppression drug (Fluorouracil,5-FU) were used to construct the blood deficiency mice model, and the red blood cells (RBC) and hemoglobin (HGB) levels were measured. Results showed that, on the 12th day, the RBC level of the bioactive peptides group has a significant difference (P<0.05) compared with the blood deficiency group; During the period of 6 to 12 days, the effect of rising the RBC and HGB level of the bioactive peptides group is significantly quicker than the Ejiao group (P<0.05). On the 18th day, the HGB level of the bioactive peptides group exceeds the other groups, and is significantly different (P<0.05) from the normal control group. The RBC level of the bioactive peptides component A group is higher than the blood deficiency group, without significant difference (P>0.05), while the blood deficiency group and the Ejiao group are significantly different (P<0.01) from the normal control group. On the 20th day, there is no significant difference (P>0.05) between the bioactive peptides component A group and the normal control group on the level of HGB, while the blood deficiency group and the Ejiao group are significantly different (P<0.01) from the normal control group.
2. To observe the anti-inflammatory activity of the TBSF peptide, the carrageenan-induced paw edema in mice model, ear xylene cause inflammation model and capillary permeability model were used in the research. The result showed that, the peptides of TBSF could inhibit the synthesis or releases of PGE2 in body tissues significantly, but to the apparent edema of body, no significant inhibit effect was found. Those suggest that the peptides of TBSF exhibited anti-inflammatory activities to some extent.
3. Investigations on the analgesic effects were conducted, including hot-plate test model and acetic acid-induced writhing test model. The results showed that the low and high doses of TBSF peptides did not increase the latency time of mice significantly, only a weak effect of analgesic was showed, but the peptides could significantly decreased the number of writhing, and inhibited the writhing response by 68.4%, showed their analgesic effects to chemical injury. To the hot-plate test model, the analgesic effect was not so significant, a low analgesic tendency exhibited. Those suggest that TBSF peptide has a better analgesic effect to chemical injury, rather than the thermal pain stimulus.
4. To study on the liver protective effect of TBSF peptides, CCI4 and D-galactosamine induced hepatic injury model were established respectively. The results were found that the ALT, AST, MDA, LI, and SI indexes were not significantly inhibited, and the SOD level was not increased. Those means remarkable protective effect was not showed to the liver injury, the effect on the inhibition of lipid peroxidation to body tissues was inconspicuous, and peptides did not show a prominent inhibitory action to the pharmaceuticals induced liver and spleen accretion.
引文
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