Discovery of natural products capable of inducing porcine host defense peptide gene expression using cell-based high throughput screening.
Discovery of natural products capable of inducing porcine host defense peptide gene expression using cell-based high throughput screening.
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DOI：
10.1186/s40104-020-00536-0
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发表时间：
2021-01-11
影响因子：
7
通讯作者：
Zhang G
中科院分区：
文献类型：
--
作者：
Wang J;Lyu W;Zhang W;Chen Y;Luo F;Wang Y;Ji H;Zhang G
饲料中抗生素正在全球范围内逐步淘汰。迫切需要抗生素的替代品来维持动物健康和生产性能。宿主防御肽（HDPs）具有广谱的抗菌和免疫调节功能。增强内源性HDPs的合成代表了疾病控制和预防的有前景的抗生素替代策略。为了鉴定具有刺激内源性HDP合成能力的天然产物，我们使用新建立的稳定的荧光素酶报告细胞系IPEC-J2/pBD 3-luc对1261种天然产物进行了高通量筛选。使用RT-qPCR验证命中化合物在猪IPEC-J2肠上皮细胞、3D 4/31巨噬细胞和空肠外植体中诱导HDP基因的能力。增强猪3D 4/31巨噬细胞对革兰氏阴性菌（产肠球菌E.大肠杆菌）和革兰氏阳性菌（金黄色葡萄球菌）的感染。在高通量筛选后，共鉴定出48种最小Z分数为2.0的天然产物，其中21种化合物在后续剂量反应实验中使荧光素酶活性增加至少2倍。进一步发现黄腐酚和脱氧紫草素在诱导pBD 3 mRNA表达方面是最有效的，在IPEC-J2、3D 4/31细胞和空肠外植体中显示出至少10倍的增加。其他化合物，如异土大黄素和毛蕊异黄酮也提高了至少10倍的pBD 3 mRNA表达在IPEC-J2细胞和空肠外植体，但不是3D 4/31细胞。除了pBD 3，其他猪HDP基因，如pBD 2，PG 1 -5，和pEP 2C诱导不同程度的黄腐酚，脱氧紫草素，异根皮苷元，和毛蕊异黄酮，虽然明确的基因和细胞类型特异性的调控模式进行了观察。理想地，这四种化合物对几种代表性炎性细胞因子基因的表达具有最小的影响。在诱导浓度下，这些化合物对3D 4/31巨噬细胞的直接抗菌活性不明显，但对革兰氏阴性菌和革兰氏阳性菌的抗菌活性有显著增强作用（P < 0.05）。我们的研究结果表明，这些新发现的天然HDP诱导化合物有可能被开发为抗生素的新替代品，用于预防和治疗家畜生产中的传染病。
In-feed antibiotics are being phased out in livestock production worldwide. Alternatives to antibiotics are urgently needed to maintain animal health and production performance. Host defense peptides (HDPs) are known for their broad-spectrum antimicrobial and immunomodulatory capabilities. Enhancing the synthesis of endogenous HDPs represents a promising antibiotic alternative strategy to disease control and prevention. To identify natural products with an ability to stimulate the synthesis of endogenous HDPs, we performed a high-throughput screening of 1261 natural products using a newly-established stable luciferase reporter cell line known as IPEC-J2/pBD3-luc. The ability of the hit compounds to induce HDP genes in porcine IPEC-J2 intestinal epithelial cells, 3D4/31 macrophages, and jejunal explants were verified using RT-qPCR. Augmentation of the antibacterial activity of porcine 3D4/31 macrophages against a Gram-negative bacterium (enterotoxigenic E. coli) and a Gram-positive bacterium (Staphylococcus aureus) were further confirmed with four selected HDP-inducing compounds. A total of 48 natural products with a minimum Z-score of 2.0 were identified after high-throughput screening, with 21 compounds giving at least 2-fold increase in luciferase activity in a follow-up dose-response experiment. Xanthohumol and deoxyshikonin were further found to be the most potent in inducing pBD3 mRNA expression, showing a minimum 10-fold increase in IPEC-J2, 3D4/31 cells, and jejunal explants. Other compounds such as isorhapontigenin and calycosin also enhanced pBD3 mRNA expression by at least 10-fold in both IPEC-J2 cells and jejunal explants, but not 3D4/31 cells. In addition to pBD3, other porcine HDP genes such as pBD2, PG1-5, and pEP2C were induced to different magnitudes by xanthohumol, deoxyshikonin, isorhapontigenin, and calycosin, although clear gene- and cell type-specific patterns of regulation were observed. Desirably, these four compounds had a minimum effect on the expression of several representative inflammatory cytokine genes. Furthermore, when used at HDP-inducing concentrations, these compounds showed no obvious direct antibacterial activity, but significantly augmented the antibacterial activity of 3D4/31 macrophages (P < 0.05) against both Gram-negative and Gram-positive bacteria. Our results indicate that these newly-identified natural HDP-inducing compounds have the potential to be developed as novel alternatives to antibiotics for prophylactic and therapeutic treatment of infectious diseases in livestock production.
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影响因子：
2.6
作者：
Moravej, Hoda;Moravej, Zahra;Mirnejad, Reza
通讯作者：
Mirnejad, Reza
影响因子：
6.1
作者：
Chu, Xin Yi;Yang, Shi Zhen;Wu, Jiang Wei
通讯作者：
Wu, Jiang Wei
DOI：
10.12945/j.aorta.2016.16.014
发表时间：
2016-08-01
期刊：
Aorta (Stamford, Conn.)
影响因子：
--
作者：
Curtis, Alexander E;Smith, Tanya A;Elefteriades, John A
通讯作者：
Elefteriades, John A
影响因子：
7.5
作者：
Ma, Ran;Yuan, Fang;Wang, Fulai
通讯作者：
Wang, Fulai
DOI：
10.1016/j.bbamcr.2016.07.005
发表时间：
2016-11-01
影响因子：
5.1
作者：
Jeung, Yun-Ji;Kim, Han-Gyeul;Chung, Kyung-Sook
通讯作者：
Chung, Kyung-Sook