Yu Yihao, Song Qingyang, Li Hongmei, Wang Shujing, Zhao Xiaomin, Zhao Ningning, Zhang Xiao
Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Shandong Agricultural University, Tai'an, China.
Shandong Provincial Key Laboratory of Zoonoses, Shandong Agricultural University, Tai'an, China.
Acta Parasitol. 2025 Jun 11;70(3):130. doi: 10.1007/s11686-025-01063-6.
Recent years have seen increased focused on developing new anti-parasitic drugs that are both highly effective and low in toxicity, as the widespread use of existing anti-parasitic drugs has raised growing concerns. Natural products have gained significant interest due to their diverse biological activities with minimal toxic side effects. Our previous studies have already demonstrated the good anti-E. tenella effect of Perillyl Alcohol. To further investigate its efficacy against other protozoa, we selected Toxoplasma gondii as the researchsubject.
In this study, we utilized the CCK-8 assay in vitro to assess the cytotoxicity of Perillyl Alcohol on DF-1 cells. The impact of Perillyl Alcohol of T. gondii tachyzoite invasion and intracellular proliferation were investigated in vitro. In vivo, we evaluated the effect of Perillyl Alcohol on the pathogenicity of T. gondii, including host survival time, liver and spleen tissue damage, cysts formed in the brain. Furthermore, we analyzed the expression levels of isoprenylation-related genes using quantitative PCR (qPCR).
The half maximal inhibitory concentration (CC50) of Perillyl Alcohol against DF-1 cells was determined to be 525.0. In vitro studies showed that treatment with Perillyl Alcohol effectively inhibited the invasion rate and intracellular proliferation of T. gondii tachyzoite. The half maximal inhibitory concentration (IC50) was calculated to be 314.3, and further analysis yielded a selectivity index (SI) of 1.67. In vivo, Perillyl Alcohol treatment prolonged the survival time and increased the survival rate of T. gondii-infected mice, while reducing the parasite burden in liver and spleen tissues. It also demonstrated a certain protective effect against T. gondii-induced tissue damage, including effectively alleviating hepatosplenomegaly and mitigating the elevation of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels induced by hepatic injury. Building on this foundation, we further explored the impact of Perillyl Alcohol on the formation of brain cysts and found that it could significantly reduce the number of brain cysts induced by the Pru strain of T. gondii infection. After treatment with Perillyl Alcohol, the expression levels of isoprenylation-related enzymes Tgmecs, Tgdxr, and Tghdr were significantly reduced.
These results demonstrate that Perillyl Alcohol may suppress the growth of T. gondii by significantly inhibiting the expression of enzymes involved in isoprenoid biosynthesis.
近年来，随着现有抗寄生虫药物的广泛使用引发了越来越多的关注，人们越来越注重开发高效低毒的新型抗寄生虫药物。天然产物因其具有多样的生物活性且毒副作用极小而备受关注。我们之前的研究已经证明了紫苏醇对柔嫩艾美耳球虫具有良好的抗虫效果。为了进一步研究其对其他原生动物的疗效，我们选择了刚地弓形虫作为研究对象。
在本研究中，我们利用体外CCK-8法评估紫苏醇对DF-1细胞的细胞毒性。体外研究紫苏醇对刚地弓形虫速殖子入侵和细胞内增殖的影响。在体内，我们评估了紫苏醇对刚地弓形虫致病性的影响，包括宿主存活时间、肝脏和脾脏组织损伤、脑内形成的囊肿。此外，我们使用定量PCR（qPCR）分析异戊二烯化相关基因的表达水平。
紫苏醇对DF-1细胞的半数最大抑制浓度（CC50）测定为525.0。体外研究表明，紫苏醇处理可有效抑制刚地弓形虫速殖子的入侵率和细胞内增殖。计算得出半数最大抑制浓度（IC50）为314.3，进一步分析得出选择性指数（SI）为1.67。在体内，紫苏醇处理延长了感染刚地弓形虫小鼠的存活时间并提高了存活率，同时降低了肝脏和脾脏组织中的寄生虫负荷。它还对刚地弓形虫诱导的组织损伤表现出一定的保护作用，包括有效减轻肝脾肿大以及减轻肝损伤诱导的血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平的升高。在此基础上，我们进一步探究了紫苏醇对脑囊肿形成的影响，发现它可以显著减少由刚地弓形虫Pru株感染诱导的脑囊肿数量。用紫苏醇处理后，异戊二烯化相关酶Tgmecs、Tgdxr和Tghdr的表达水平显著降低。
这些结果表明，紫苏醇可能通过显著抑制类异戊二烯生物合成中相关酶的表达来抑制刚地弓形虫的生长。
