从医用水蛭 Whitmania pigra 中分离得到的动脉粥样硬化中巨噬细胞迁移的肽抑制剂。
A peptide inhibitor of macrophage migration in atherosclerosis purified from the leech Whitmania pigra.
机构信息
Marine College, Shandong University, Weihai, 264209, PR China; Weihai International Biotechnology Research and Development Centre, Shandong University Weihai, 264209, PR China.
Marine College, Shandong University, Weihai, 264209, PR China; Weihai International Biotechnology Research and Development Centre, Shandong University Weihai, 264209, PR China.
出版信息
J Ethnopharmacol. 2020 May 23;254:112723. doi: 10.1016/j.jep.2020.112723. Epub 2020 Feb 28.
ETHNOPHARMACOLOGICAL RELEVANCE
Atherosclerosis has become a worldwide public health problem that seriously threatens human health. Leech is traditional Chinese medicine that can be utilized to treat cardiovascular disease. Based on the anti-atherosclerosis activity of leech hydrolysate, we separated and purified the leech peptide capable of inhibiting macrophage migration and studied the pathways of the anti-migration leech peptide.
MATERIALS AND METHODS
The leech peptide capable of inhibiting macrophage migration that measured by cell migration assays from the leech Whitmania pigra was separated and purified by Q Sepharose FF strong alkaline anion exchange column chromatography, Superdex 30, Superdex peptide and G10 gel column chromatography. And the purity, molecular weight of the leech peptide was determined by high-performance liquid chromatography and high-resolution mass spectrometry. The pathways of anti-migration to macrophages of the leech peptide were studied by inhibitors, Western blotting and RT-PCR.
RESULTS
We obtained a purified leech peptide with a sequence of EAGSAKELEGDPVAG from the leech Whitmania pigra. We also showed that the anti-migration to macrophages of the leech peptide was blocked by c-Jun N-terminal kinase (JNK) inhibitor and p38 mitogen-activated protein kinase (p38 MAPK) inhibitor. Moreover, the result of RT-PCR and Western blotting revealed that the leech peptide induced an increase in JNK, p38 phosphorylation and the transcription of mitogen-activated protein kinase kinase kinase 4 (MEKK4) and apoptosis signal-regulating kinase 2 (ASK2). These data indicated that the anti-migration to macrophages of the leech peptide occurred through JNK and p38 MAPK pathways. In addition, the results demonstrated that the leech peptide had no significant effect on the immunological activity of macrophages including phagocytic ability, lysozyme activity, and levels of expression of inflammatory factors.
CONCLUSION
A sequence peptide was obtained from the hydrolysate of leech Whitmania pigra that inhibits macrophage migration.
民族药理学相关性
动脉粥样硬化已成为全球公共卫生问题，严重威胁人类健康。水蛭是一种传统的中药，可用于治疗心血管疾病。基于水蛭水解物的抗动脉粥样硬化活性，我们分离并纯化了能够抑制巨噬细胞迁移的水蛭肽，并研究了抗迁移水蛭肽的途径。
材料和方法
通过细胞迁移实验从水蛭Whitmania pigra 中分离和纯化能够抑制巨噬细胞迁移的水蛭肽，采用 Q Sepharose FF 强碱阴离子交换柱层析、Superdex 30、Superdex 肽和 G10 凝胶柱层析进行分离和纯化。通过高效液相色谱和高分辨率质谱法测定水蛭肽的纯度和分子量。通过抑制剂、Western blot 和 RT-PCR 研究了水蛭肽对巨噬细胞迁移的抑制途径。
结果
从水蛭 Whitmania pigra 中获得了一个具有 EAGSAKELEGDPVAG 序列的纯化水蛭肽。我们还表明，水蛭肽对巨噬细胞的迁移抑制作用被 c-Jun N 端激酶（JNK）抑制剂和 p38 丝裂原活化蛋白激酶（p38 MAPK）抑制剂阻断。此外，RT-PCR 和 Western blot 的结果表明，水蛭肽诱导 JNK、p38 磷酸化以及丝裂原活化蛋白激酶激酶激酶 4（MEKK4）和凋亡信号调节激酶 2（ASK2）的转录增加。这些数据表明，水蛭肽对巨噬细胞的迁移抑制作用是通过 JNK 和 p38 MAPK 途径发生的。此外，结果表明，水蛭肽对巨噬细胞的免疫活性（包括吞噬能力、溶菌酶活性和炎症因子表达水平）没有显著影响。
结论
从水蛭 Whitmania pigra 的水解物中获得了一种抑制巨噬细胞迁移的序列肽。
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