In this ex vivo study, S. cervi parasitoses were treated with Ivermectin (50 μM), Albendazole (200 μM) alone and Ivermectin + Albendazole (50 + 200 μM) at 37°C for 8 h and the motility and viability of the parasitoses were evaluated. Individually both drugs Ivermectin (Iver) and Albendazole (Alb) are reported to affect the function and integrity of ER, however till date, no reports are available on the functional changes in ER due to a combined Iver and Alb treatment of bovine helminth parasitosis. Here, we report the lethal effect of a combination treatment of Iver and Alb against adult bovine filarial parasitosis Setaria cervi. The underlying mechanism of drug action was elucidated by performing a systematic biochemical, molecular and proteomics based study. Altered calcium homeostasis in drug treated parasitoses lead to reduction in levels of total Endoplasmic Reticulum (ER) calcium by 50 % and 61 % and elevation by 50 % and 63 % in cytosol in Iver alone and Iver + Alb treated parasitoses respectively. Further, it was found that upregulated expression of ER localized GRP94, galactosyltransferase and glycosyltransferase activity in addition to reduction in activity of PDI indicated ER stress mechanisms being operative under combined drug treatment. Marked rise of 79 % reactive oxygen species and reduced antioxidant levels induced oxidative stress in drug treated parasitosis. The collective effect of both ER and oxidative stress might have triggered apoptosis, as evidenced by the elevated calpain activity, reduction of 67 % in cytochrome c oxidase and 83 % rise in caspase-3 activity in the Iver + Alb treated parasitoses respectively.

The ER proteome analysis by 2D gel electrophoresis revealed 76 spots in the control and 56 spots in the treated proteome. A MALDI-MS/MS analysis of some of the differentially expressed spots of the combination drug treated parasitoses identified glucuronosyltransferase as a major upregulated protein with a fold change of 1.81. Trafficking protein, acyl transferase, MATH involved in protein folding were also found to be downregulated. Thus, this study based on biochemical and proteomic approaches indicates that a combination of anti-filarial drugs Iver and Alb can alter calcium homeostasis in bovine filarial parasitosis leading to induction of ER stress culminating into apoptosis.

在该离体研究中，S. Cervi的寄生虫病用伊维菌素（50μM），阿苯达唑（200μM）单独和伊维菌素+阿苯达唑（50 + 200μM），在37℃处理8小时和寄生虫病的活力和活被评估。据报道，伊维菌素 (Iver) 和阿苯达唑 (Alb) 两种药物单独影响 ER 的功能和完整性，但迄今为止，尚无关于 Iver 和 Alb 联合治疗牛蠕虫寄生虫引起的 ER 功能变化的报告。在这里，我们报告了 Iver 和 Alb 联合治疗对成年牛丝虫寄生虫病狗尾草的致死作用. 通过进行系统的生化、分子和蛋白质组学研究，阐明了药物作用的潜在机制。在单独的 Iver 和 Iver + Alb 处理的寄生虫中，药物治疗的寄生虫中钙稳态的改变导致总内质网 (ER) 钙的水平分别降低 50% 和 61%，细胞溶质中的钙水平升高 50% 和 63%。此外，发现 ER 的表达上调除了 PDI 活性的降低外，还定位了 GRP94、半乳糖基转移酶和糖基转移酶活性，表明 ER 应激机制在联合药物治疗下有效。在药物治疗的寄生虫病中，79% 的活性氧物质显着增加，抗氧化剂水平降低，诱导氧化应激。ER 和氧化应激的共同作用可能触发了细胞凋亡，

通过二维凝胶电泳进行的 ER 蛋白质组分析显示，对照中有 76 个点，处理后的蛋白质组中有 56 个点。对组合药物处理的寄生虫的一些差异表达点的 MALDI-MS/MS 分析将葡萄糖醛酸转移酶鉴定为主要上调蛋白质，倍数变化为 1.81。还发现转运蛋白、酰基转移酶、参与蛋白质折叠的 MATH 被下调。因此，这项基于生化和蛋白质组学方法的研究表明，抗丝虫药物 Iver 和 Alb 的组合可以改变牛丝虫寄生虫病中的钙稳态，导致诱导内质网应激，最终导致细胞凋亡。