Anaphalis margaritacea ethanol extract exhibits potent anti-Trichinella spiralis activity via mitochondrial dysfunction and host tissue protection

Trichinellosis is a food-borne zoonotic disease caused by Trichinella spiralis, for which effective treatment of muscle larvae remains limited. Medicinal plants represent an important resource for the discovery of new anthelmintic agents against this infection. Screening of 30 medicinal plants with documented or putative antiparasitic use identified the 80% ethanol extract of Anaphalis margaritacea (AMEE) as the most active extract against T. spiralis muscle larvae. AMEE caused complete larval mortality within 72 h at 100 μg/mL and showed a clear dose-dependent larvicidal effect in vitro. To provide a preliminary phytochemical basis for this activity, UPLC-Q-TOF-MS/MS profiling was performed, and eight major constituents were tentatively identified in the active AMEE-derived fraction, mainly belonging to organic acid and flavonoid classes. In a murine model of infection, oral administration of AMEE significantly reduced muscle larval burdens by up to 59.7% in a dose-responsive manner. In parallel with parasite reduction, AMEE treatment alleviated parasite-induced muscle fibre damage and inflammatory infiltration, accompanied by decreased expression of Th2-associated cytokines (IL-6 and IL-13) and TGF-β, as well as normalisation of oxidative stress–related genes, including Nqo1 and Sod2. To elucidate the mechanism underlying the anthelmintic activity, integrated analyses combining proteomics, mitochondrial enzyme activity assays and transmission electron microscopy were performed. AMEE exposure markedly suppressed the activities of mitochondrial respiratory complexes I, II and IV, reduced complex V activity, and induced mitochondrial swelling and cristae disruption in muscle larvae, consistent with impaired bioenergetic function, reduced ATP production and larval death. Together, these findings demonstrate that A. margaritacea ethanol extract exerts pronounced anti-T. spiralis activity in vitro and in vivo, associated with mitochondrial dysfunction in muscle larvae and attenuation of infection-induced muscle pathology.

旋毛虫病（Trichinellosis）是由旋毛形线虫（Trichinella spiralis）引发的食源性人畜共患病，目前针对其肌肉幼虫的有效治疗方案仍较为有限。药用植物是开发针对该感染的新型抗蠕虫药物（anthelmintic agents）的重要资源。研究团队对30种经文献记载或推测具备抗寄生虫活性的药用植物开展筛选，最终确定珠光香青（Anaphalis margaritacea）的80%乙醇提取物（AMEE）为抗旋毛形线虫肌肉幼虫活性最强的提取物。AMEE在100 μg/mL浓度下可在72小时内完全杀灭幼虫，且在体外实验中呈现出明确的剂量依赖性杀幼虫效应。为明确该活性的初步植物化学基础，研究人员采用超高效液相色谱-串联四极杆飞行时间质谱（UPLC-Q-TOF-MS/MS）开展成分表征，从活性AMEE组分中初步鉴定出8种主要成分，主要隶属于有机酸类与黄酮类化合物家族。在旋毛虫感染小鼠模型（murine model）中，口服AMEE可呈剂量依赖性显著降低肌肉幼虫负荷，最高降幅可达59.7%。伴随寄生虫负荷降低的同时，AMEE治疗可减轻寄生虫诱导的肌肉纤维损伤与炎症浸润，同时下调Th2型相关细胞因子（Th2-associated cytokines，包括IL-6和IL-13）及转化生长因子β（TGF-β）的表达水平，并使包括Nqo1、Sod2在内的氧化应激相关基因（oxidative stress–related genes）表达恢复至正常水平。为阐明其抗蠕虫活性的潜在作用机制，研究团队整合蛋白质组学（proteomics）、线粒体酶活性检测及透射电子显微镜（transmission electron microscopy）分析开展联合研究。实验结果显示，AMEE处理可显著抑制线粒体呼吸链复合物（mitochondrial respiratory complexes）I、II、IV的活性，降低复合物V的活性，并诱导肌肉幼虫出现线粒体肿胀及嵴膜破坏，这与生物能量功能受损、三磷酸腺苷（ATP）生成减少及幼虫死亡的现象高度吻合。综上，本研究证实珠光香青乙醇提取物在体外及体内均表现出显著的抗旋毛形线虫活性，其作用机制与肌肉幼虫线粒体功能障碍及感染诱导的肌肉病理损伤缓解密切相关。