蝉蛹肽的制备及其免疫调节活性变化
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石家庄以岭药业股份有限公司

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国家重点研发计划“中医药现代化”重点专项课题


Preparation of Peptides from Cryptotympana atrata and its Immunomodulatory Activity changes
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Shijiazhuang Yiling Pharmaceutical Co., Ltd.

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    摘要:

    该文对蝉蛹肽(Cryptotympana Atrata Peptide, CA-Peptide)的制备工艺进行优化,并比较CA-Peptide和蝉蛹蛋白(Cryptotympana Atrata Protein, CA-Protein)的免疫调节活性。最佳工艺:碱性蛋白酶37071和胰蛋白酶复合酶解,加酶量各4%(m/m),底物浓度5%(m/m)、初始pH值10.5、酶解温度50℃,两种蛋白酶先后酶解1.5 h。动物试验显示,CA-Peptide和CA-Protein均能不同程度改善环磷酰胺所致免疫低下小鼠的相关指标。相同剂量下,CA-Peptide组小鼠的脾脏指数和脾淋巴细胞增值率分别为2.40 mg/g和51.07%,显著(P<0.05)高于CA-Protein组;CA-Peptide组小鼠的血清免疫球蛋白G、免疫球蛋白A、免疫球蛋白M、肿瘤坏死因子α、白细胞介素-4和白细胞介素-10含量分别为701.67、299.19、239.88、20.11、33.86和71.15 ng/L,显著(P<0.05)高于CA-Protein组;CA-Peptide组小鼠的胸腺指数和血清γ干扰素含量分别为1.36 mg/g和36.26 ng/L,显著(P<0.05)低于CA-Protein组;CA-Peptide组小鼠的自然杀伤细胞活性及血清白细胞介素-2含量与CA-Protein组无显著性差异(P>0.05)。结论,酶解工艺显著改变了CA-Protein的免疫调节活性,产物CA-Peptide对多项免疫指标的改善作用优于CA-Protein。

    Abstract:

    In this paper, the preparation process of Cryptotympana Atrata Peptide (CA-Peptide) was optimized, and the immunomodulatory activities of CA-Peptide and Cryptotympana Atrata Protein (CA-protein) were compared. The optimal preparation process: 4% alkaline proteinase 37071 (m/m) and 4% trypsin enzymolysis (m/m), 5% substrate concentration (m/m), initial pH 10.5, enzyme temperature 50℃, two kinds of protease enzymolysis for 1.5 hours respectively. Animal experiments showed that the related indexes of immunocompromised mice induced by cyclophosphamide all could be improved in different degrees by CA-Peptide and CA-Protein. At the same dose, the spleen index and proliferation rate of splenic lymphocytes in CA-Peptide group (2.40 mg/g and 51.07%, respectively) were significantly (P<0.05) higher than those of CA-Protein group. The immunoglobulin G, immunoglobulin A, immunoglobulin M, tumor necrosis factor-α, interleukin-4 and interleukin-10 in CA-Peptide group (701.67, 299.19, 239.88, 20.11, 33.86 and 71.15 ng/L, respectively) were significantly (P<0.05) higher than those of CA-Protein group. The thymus index and serum interferon-γ in CA-Peptide group (1.36 mg/g and 36.26 ng/L, respectively) were significantly (P<0.05) lower than those of CA-Protein group. There was no significant difference in natural killer cell activity and serum interleukin-2 level between CA-Peptide group and CA-Protein group (P > 0.05). In conclusion, the enzymatic hydrolysis process significantly changes immunomodulatory activity of CA-Protein, and the product CA-Peptide has better improvement on many immune indexes than CA-Protein.

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  • 收稿日期:2024-05-23
  • 最后修改日期:2024-08-26
  • 录用日期:2024-08-29
  • 在线发布日期: 2025-05-06
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