Ali Muhammad Muddassir, Farhad Zainab, Wasim Muhammad, Raza Sohail, Almutairi Mikhlid H, Zahra Kainat, Saleem Muhammad Usman, Mehmood Khalid
Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
Institute of Microbiology, University of Veterinary and Animal Sciences, Lahore, Pakistan.
PLoS One. 2024 May 3;19(5):e0296255. doi: 10.1371/journal.pone.0296255. eCollection 2024.
Ivermectin (IVM) is an anti-parasitic drug which is used for treating parasitic infestations. It has been used in humans for treating intestinal strongyloidiasis and onchocerciasis however, currently researchers are investigating its potential for treating coronavirus SARS-CoV-2. Due to its broad-spectrum activities, IVM is being used excessively in animals which has generated an interest for researchers to investigate its toxic effects. Cytotoxic and genotoxic effects have been reported in animals due to excessive usage of IVM. Therefore, this study aims to evaluate the cytotoxic and genotoxic effects of IVM on the Madin-Darby-Bovine-Kidney (MDBK) cell line by examining the expression of a DNA damage-responsive gene (OGG1). Cytotoxicity of IVM was tested using an assay (MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), whereas the genotoxicity was evaluated using comet assay along with micronucleus assay. Moreover, the gene expression of DNA damage response gene (OGG1) was measured by qRT-PCR, after extraction of RNA from the MDBK cell line using the TRIzol method and its conversion to cDNA by reverse-transcriptase PCR. During the experiment, cell viability percentage was measured at different doses of IVM i.e., 25%, 50%, 75%, along with LC50/2, LC50 and LC50*2. It was observed that the gene expression of OGG1 increased as the concentration of IVM increased. It was concluded that IVM has both cytotoxic and genotoxic effects on the MDBK cell line. Furthermore, it is recommended that studies related to the toxic effects of IVM at molecular level and on other model organisms should be conducted to combat its hazardous effects.
伊维菌素（IVM）是一种抗寄生虫药物，用于治疗寄生虫感染。它已被用于人类治疗肠道类圆线虫病和盘尾丝虫病，然而，目前研究人员正在研究其治疗冠状病毒SARS-CoV-2的潜力。由于其具有广谱活性，IVM在动物中被过度使用，这引起了研究人员对其毒性作用进行研究的兴趣。据报道，由于IVM的过度使用，动物出现了细胞毒性和基因毒性作用。因此，本研究旨在通过检测DNA损伤反应基因（OGG1）的表达，评估IVM对马-达-比牛肾（MDBK）细胞系的细胞毒性和基因毒性作用。使用一种检测方法（MTT 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）检测IVM的细胞毒性，而使用彗星试验和微核试验评估基因毒性。此外，在使用TRIzol方法从MDBK细胞系中提取RNA并通过逆转录酶PCR将其转化为cDNA后，通过qRT-PCR测量DNA损伤反应基因（OGG1）的基因表达。在实验过程中，在不同剂量的IVM下测量细胞活力百分比，即25%、50%、75%，以及LC50/2、LC50和LC50*2。观察到随着IVM浓度的增加，OGG1的基因表达增加。得出的结论是，IVM对MDBK细胞系具有细胞毒性和基因毒性作用。此外，建议开展与IVM在分子水平和其他模式生物上的毒性作用相关的研究，以对抗其有害影响。
