Antimicrobial peptide temporin-1CEa isolated from frog skin secretions inhibits the proinflammatory response in lipopolysaccharide-stimulated RAW264.7 murine macrophages through the MyD88-dependent
Antimicrobial peptide temporin-1CEa isolated from frog skin secretions inhibits the proinflammatory response in lipopolysaccharide-stimulated RAW264.7 murine macrophages through the MyD88-dependent
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从青蛙皮肤分泌物中分离出的抗菌肽 temporin-1CEa 通过 MyD88 依赖性信号通路抑制脂多糖刺激的 RAW264.7 小鼠巨噬细胞的促炎反应
DOI：
10.1016/j.molimm.2021.01.007
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发表时间：
2021
影响因子：
3.6
通讯作者：
Dejing Shang
中科院分区：
文献类型：
--
作者：
Juan Zhang;Yue Sun;Yao Kang;Dejing Shang
Temporin-1CEa是从中国林蛙（Rana chensinensis）皮肤分泌物中分离得到的一种蛋白质，对革兰氏阳性菌和革兰氏阴性菌具有广谱抗菌活性，并具有抗肿瘤活性。LK2（6）和LK2（6）A（L）是通过替换氨基酸获得的temporin-1CEa的类似物，并显示出改善的抗癌活性。本研究探讨了temporin-1CEa及其类似物LK2（6）和LK2（6）A（L）在脂多糖（LPS）刺激的小鼠RAW 264.7巨噬细胞中的抗炎活性及其作用机制。结果表明，Temporin-1 CEa及其类似物通过抑制核因子-κ B和丝裂原活化蛋白激酶的蛋白表达以及MyD 88依赖的信号通路，抑制细胞因子肿瘤坏死因子-α和白细胞介素-6的产生。等温滴定量热法研究表明，temporin-1CEa、LK2（6）和LK2（6）A（L）与LPS（一种重要的炎症诱导剂）具有亲和力，Kd值分别为0.1、0.03和0.06 μ M。圆二色性和zeta电位实验表明，temporin-1CEa及其类似物通过带正电荷的多肽和带负电荷的LPS之间的静电结合与LPS相互作用，从而中和LPS的毒性。
Temporin-1CEa, which is isolated from the skin secretions of the Chinese brown frogRana chensinensis, exhibits broad-spectrum antimicrobial activity against gram-positive and gram-negative bacteria and antitumor activity. LK2(6) and LK2(6)A(L) are the analogs of temporin-1CEa obtained by replacing amino acids and displayed an improved anticancer activity. In the present study, the anti-inflammatory activity and mechanism of action of temporin-1CEa and its analogs LK2(6) and LK2(6)A(L) in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages were investigated. The results showed that temporin-1CEa and its analogs decreased the production of the cytokines tumor necrosis factor-α and interleukin-6 by inhibiting the protein expression of nuclear factor-κB and mitogen-activated protein kinase and the MyD88-dependent signaling pathway. Isothermal titration calorimetry studies revealed that temporin-1CEa, LK2(6) and LK2(6)A(L) exhibited binding affinities to LPS, an important inflammatory inducer, withKdvalues of 0.1, 0.03 and 0.06 μM, respectively. Circular dichroism and zeta potential experiments showed that temporin-1CEa and its analogs interacted with LPS by electrostatic binding between the positively charged peptides and negatively charged LPS, resulting in the neutralization of LPS toxicity.