Determination of anti-cancer and antioxidant properties of protein extracts obtained from aquatic Helophorus (Coleoptera: Helophoridae) insects

Tuba Elhazar, B. Kaya, F. Caf
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Abstract

In this study, protein extraction was performed from the aquatic insect Helophorus aquaticus (Linnaeus, 1758) and Helophorus syriacus (Kuwert, 1885) species belonging to the genus Helophorus (Coleoptera: Helophoridae). Then, these protein extracts were studied in vitro. These species were collected from the shallow parts of various streams, springs, creeks, ponds and hot springs from Bingöl city centre and its districts between May to June 2017. The protein amount was determined 34.78 mg/mL in the H. aquaticus and 35.14 mg/mL in the H. syriacus, after that he antioxidant capacity of protein extracts was examined. Metal chelating activity was determined as 90-88.5% and DPPH removal activity 53.19-61.7% for H. aquaticus and H. syriacus respectively. Protein samples belonging to both species were tested for cell vitality with WST-1 in PC-3 (prostate cancer) cells with in vitro cell culture. Upon examination of the test results, it has been found out that protein extracts from both of the studied species caused a decrease in cell inhibition. The highest cell inhibition was observed in samples with 1000 μg/mL insect protein extract added. In this study, protein expression providing apoptosis was examined with the Western blot technique after the effective dose was established. By looking at the proteins of Cyt-C and Caspase 3 with the Western blot technique, the efficacy of the protein extracts from both species was demonstrated effectively for the in vitro PC-3 line in non-apoptosis cell death. As a result of the study, insect proteins were shown to support the production of proteins that ensure cell death with the western blot technique.
水生Helophorus(鞘翅目:helophordae)昆虫蛋白质提取物抗癌和抗氧化性能的测定
本研究对Helophorus aquaticus (Linnaeus, 1758)和Helophorus syriacus (Kuwert, 1885)属Helophorus属水生昆虫(鞘翅目:Helophoridae)进行了蛋白质提取。然后,对这些蛋白提取物进行体外研究。这些物种是在2017年5月至6月期间从Bingöl市中心及其地区的各种溪流、温泉、小溪、池塘和温泉的浅水部分收集的。测定了水杨中蛋白质含量为34.78 mg/mL,叙红中蛋白质含量为35.14 mg/mL,并测定了蛋白质提取物的抗氧化能力。结果表明,水草和红草的金属螯合活性为90 ~ 88.5%,对DPPH的去除活性为53.19 ~ 61.7%。在体外细胞培养的PC-3(前列腺癌)细胞中,用WST-1检测了这两个物种的蛋白质样本的细胞活力。在检查测试结果后,发现两种被研究物种的蛋白质提取物导致细胞抑制减少。添加1000 μg/mL的昆虫蛋白提取物对细胞的抑制作用最强。在本研究中,在有效剂量确定后,用Western blot技术检测凋亡相关蛋白的表达。通过Western blot技术观察Cyt-C和Caspase 3蛋白表达,证实了两种蛋白提取物对体外PC-3细胞系非凋亡细胞死亡的有效作用。作为研究的结果,昆虫蛋白被证明支持蛋白质的生产,确保细胞死亡与western blot技术。
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