Tang Yaqin, Hou Shengjie, Li Xianyao, Wu Mengqi, Ma Binbin, Wang Zheng, Jiang Jinying, Deng Meichun, Duan Zhigui, Tang Xing, Liu Yuan, Wang Wenhua, Han Xiaoqing, Jiang Liping
Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha, Hunan, 410013, PR China.
The First Department of General Surgery, The Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, PR China.
Exp Parasitol. 2019 Mar;198:17-25. doi: 10.1016/j.exppara.2019.01.009. Epub 2019 Jan 22.
Toxoplasmosis is a widely distributed parasitic protozoan disease, caused by Toxoplasma gondii (T. gondii). High prevalence of toxoplasmosis and limitations of conventional treatments lead to a search for new therapeutic drugs. Lycosin-I is a linear peptide, derived from the venom of the spider Lycosa singoriensis. The aim of the present study was to determine the anti-parasitic effect of lycosin-Ι against T. gondii. In vitro, the anti-T. gondii activities of lycosin-Ι were evaluated by MTT assay, trypan blue exclusion assay, cell counting assay and plaque assay. Cytokines of IL-6 and IL-8 were measured by quantitative PCR. In addition, the structures of tachyzoites treated with lycosin-Ι were also observed by scanning and transmission electron microscopy. In vivo, mice were challenged with parasites treated by lycosin-I. The results revealed that lycosin-Ι had shown a significant ability to inhibit T. gondii invasion and proliferation. Cytokines of IL-6 and IL-8 were reduced by lycosin-Ι at transcription level in human foreskin fibroblast (HFF) cells infected with T. gondii tachyzoites, but they were increased compared to non-infected cells. For tachyzoites, lycosin-Ι induced their cell membrane alterations with formation of invaginations, some of them appeared to be vacuolated in their cytoplasm. Moreover, lycosin-Ι had prolonged the survival time of mice by controlling T. gondii proliferation. In conclusion, our present study provides the first evidence for anti-T. gondii by using the spider peptide lycosin-Ι. These findings suggest that lycosin-Ι is a potential alternative agent for the treatment of toxoplasmosis.
弓形虫病是一种广泛传播的寄生原生动物疾病,由刚地弓形虫(T. gondii)引起。弓形虫病的高流行率和传统治疗方法的局限性促使人们寻找新的治疗药物。狼蛛毒素-I是一种线性肽,源自狼蛛Lycosa singoriensis的毒液。本研究的目的是确定狼蛛毒素-I对刚地弓形虫的抗寄生虫作用。在体外,通过MTT法、台盼蓝排斥试验、细胞计数试验和噬斑试验评估狼蛛毒素-I的抗刚地弓形虫活性。通过定量PCR检测白细胞介素-6(IL-6)和白细胞介素-8(IL-8)的细胞因子。此外,还通过扫描电子显微镜和透射电子显微镜观察了用狼蛛毒素-I处理的速殖子的结构。在体内,用经狼蛛毒素-I处理的寄生虫攻击小鼠。结果表明,狼蛛毒素-I具有显著抑制刚地弓形虫侵袭和增殖的能力。在感染刚地弓形虫速殖子的人包皮成纤维细胞(HFF)中,狼蛛毒素-I在转录水平上降低了IL-6和IL-8的细胞因子,但与未感染细胞相比有所增加。对于速殖子,狼蛛毒素-I诱导其细胞膜改变并形成内陷,其中一些在细胞质中似乎出现空泡化。此外,狼蛛毒素-I通过控制刚地弓形虫的增殖延长了小鼠的存活时间。总之,我们的研究首次提供了利用蜘蛛肽狼蛛毒素-I抗刚地弓形虫的证据。这些发现表明,狼蛛毒素-I是治疗弓形虫病的潜在替代药物。