Wang Ningyuan, Huang Yan, Li An, Jiang Hailong, Wang Jie, Li Jianzhong, Qiu Lei, Li Ka, Lu Yiming
School of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou 350108, China.
Department of Biochemical Pharmacy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
Int J Mol Sci. 2016 Nov 22;17(11):1940. doi: 10.3390/ijms17111940.
Tumor necrosis factor (TNF)-α is a pleiotropic cytokine with intense pro-inflammatory and immunomodulatory properties, and anti-TNF-α biologics are effective therapies for various inflammatory diseases such as inflammatory bowel disease (IBD) and sepsis. Snake venom, as a traditional Chinese medicine, has been used in the treatment of inflammatory diseases in China for centuries. In this research, we constructed a venom gland T7 phage display library of the sea snake to screen bioactive compounds that antagonize TNF-α and identified a novel nine-amino-acid peptide, termed hydrostatin-TL1 (H-TL1). In enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) analyses, H-TL1 inhibited the interaction between TNF-α and TNF receptor 1 (TNFR1). Further, H-TL1 attenuated the cytotoxicity of TNF-α in L929 cells as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. H-TL1 also decreased the mRNA expression of TNF-α/TNFR1 downstream targets and suppressed the phosphorylation of well-characterized proteins of downstream signal transduction pathways in HEK-293 cells. In vivo data demonstrated that H-TL1 protects animals against dextran sodium sulfate (DSS)-induced acute colitis and lipopolysaccharide (LPS)-induced acute shock. Given its significant anti-inflammatory activity in vitro and in vivo, H-TL1 is a potential peptide for the development of new agents to treat TNF-α-associated inflammatory diseases.
肿瘤坏死因子（TNF）-α是一种具有强烈促炎和免疫调节特性的多效性细胞因子，抗TNF-α生物制剂是治疗各种炎症性疾病（如炎症性肠病（IBD）和脓毒症）的有效疗法。蛇毒作为一种传统中药，在中国用于治疗炎症性疾病已有数百年历史。在本研究中，我们构建了海蛇毒腺T7噬菌体展示文库，以筛选拮抗TNF-α的生物活性化合物，并鉴定出一种新型九氨基酸肽，命名为hydrostatin-TL1（H-TL1）。在酶联免疫吸附测定（ELISA）和表面等离子体共振（SPR）分析中，H-TL1抑制了TNF-α与TNF受体1（TNFR1）之间的相互作用。此外，通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）测定法确定，H-TL1减弱了TNF-α在L929细胞中的细胞毒性。H-TL1还降低了TNF-α/TNFR1下游靶点的mRNA表达，并抑制了HEK-293细胞中下游信号转导通路特征明确的蛋白质的磷酸化。体内数据表明，H-TL1可保护动物免受葡聚糖硫酸钠（DSS）诱导的急性结肠炎和脂多糖（LPS）诱导的急性休克。鉴于其在体外和体内均具有显著的抗炎活性，H-TL1是开发治疗TNF-α相关炎症性疾病新药的潜在肽。
