El-Garawani Islam, El-Seedi Hesham, Khalifa Shaden, El Azab Islam H, Abouhendia Marwa, Mahmoud Shaymaa
Department of Zoology, Faculty of Science, Menoufia University, Menoufia 32511, Egypt.
Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, S-10691 Stockholm, Sweden.
Pharmaceutics. 2020 Nov 19;12(11):1111. doi: 10.3390/pharmaceutics12111111.
The usage of insects as a sustainable and functional natural products resource is a new promise in complementary and alternative medicine. The present study aimed to investigate the ability of (housefly) larval hemolymph (insect blood) to display the enhanced in vitro antioxidant and cytotoxic effects. The oxidative stress (OS) was elicited by inducing lipopolysaccharides (LPS) treatment as an exogenous stressor. Determination of superoxide dismutase 1 (SOD1), glutathione (GSH), malondialdehyde (MDA) and total antioxidant capacity (TAC), and mRNA and protein expressions of SOD1, was investigated as confirmatory markers of oxidative stress induction. Cytotoxicity on cancerous MCF-7 and normal Vero cells were also evaluated using an MTT assay at 24 h post-injection. The injection of LPS induced a significant ( < 0.05) increase in SOD, GSH and TAC, whereas, the MDA was diminished. Hemolymph was collected from normal and treated larvae after 6, 12 and 24 h. The superoxide dismutase (SOD1) transcripts were significantly ( < 0.05) upregulated 6 and 12 h post-treatment, while a significant downregulation was observed after 24 h. Western blot analysis showed that SOD1 was expressed in the hemolymph of the treated larvae with an increase of 1.2 folds at 6 and 12 h and 1.6 folds at 24 h relative to the control group. LPS-treated larval hemolymphs exhibited significant cytotoxicity with respect to the untreated ones against MCF-7 while Vero cells showed no cytotoxicity for both hemolymphs. The DPPH free radical scavenging activity was examined and a significant antioxidant potential potency was observed at 6 h (50% maximal inhibitory concentration (IC): 63.3 ± 3.51 µg/mL) when compared to the control larval hemolymph (IC: 611.7 ± 10.41 µg/mL). Taken together, larval hemolymph exhibited enhanced antioxidant and consequently increased cytotoxic capacities under stressed conditions.
将昆虫用作可持续且具功能性的天然产物资源是补充和替代医学中的一个新希望。本研究旨在探究家蝇幼虫血淋巴(昆虫血液)展现出增强的体外抗氧化和细胞毒性作用的能力。通过诱导脂多糖(LPS)处理作为外源性应激源来引发氧化应激(OS)。测定超氧化物歧化酶1(SOD1)、谷胱甘肽(GSH)、丙二醛(MDA)和总抗氧化能力(TAC),以及SOD1的mRNA和蛋白质表达,作为氧化应激诱导的确认标志物。在注射后24小时,还使用MTT法评估了对癌细胞MCF - 7和正常Vero细胞的细胞毒性。LPS注射导致SOD、GSH和TAC显著(<0.05)增加,而MDA减少。在6、12和24小时后从正常和处理后的幼虫中收集血淋巴。处理后6小时和12小时,超氧化物歧化酶(SOD1)转录本显著(<0.05)上调,而在24小时后观察到显著下调。蛋白质印迹分析表明,SOD1在处理后幼虫的血淋巴中表达,相对于对照组,在6小时和12小时增加1.2倍,在24小时增加1.6倍。LPS处理的幼虫血淋巴对未处理的血淋巴而言,对MCF - 7表现出显著的细胞毒性,而Vero细胞对两种血淋巴均未显示细胞毒性。检测了DPPH自由基清除活性,与对照幼虫血淋巴(IC:611.7±10.41μg/mL)相比,在6小时时观察到显著的抗氧化潜力效能(50%最大抑制浓度(IC):63.3±3.51μg/mL)。综上所述,幼虫血淋巴在应激条件下表现出增强的抗氧化能力,并因此增强了细胞毒性能力。