Murugan Kadarkarai, Aruna Palanimuthu, Panneerselvam Chellasamy, Madhiyazhagan Pari, Paulpandi Manickam, Subramaniam Jayapal, Rajaganesh Rajapandian, Wei Hui, Alsalhi Mohamad Saleh, Devanesan Sandhanasamy, Nicoletti Marcello, Syuhei Ban, Canale Angelo, Benelli Giovanni
Parasitol Res. 2016 Feb;115(2):651-62. doi: 10.1007/s00436-015-4783-6.
Dengue is a mosquito-borne viral disease that has rapidly spread in all regions of the world in recent years. Female mosquitoes, mainly Aedes aegypti, transmit dengue. Approximately 3,900 million people, in 128 countries, are at risk of dengue infection. Recently, a focus has been provided on the potential of green-synthesized nanoparticles as inhibitors of the production of dengue viral envelope (E) protein in Vero cells and downregulators of the expression of dengue viral E gene. Algae are an outstanding reservoir of novel compounds, which may help in the fight against mosquito-borne diseases. In this research, silver nanoparticles (AgNP) were rapidly synthesized using a cheap extract of the alga Centroceras clavulatum. AgNP were characterized by UV–vis spectrophotometry, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), and X-ray diffraction (XRD). In mosquitocidal assays, LC50 values of C. clavulatum extract against A. aegypti larvae and pupae were 269.361 ppm (larva I), 309.698 ppm (larva II), 348.325 ppm (larva III), 387.637 ppm (larva IV), and 446.262 ppm (pupa). C. clavulatum extract also exhibited moderate antioxidant activity, both in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (NO) radical scavenging assays. LC50 values of C. clavulatum-synthesized AgNP were 21.460 ppm (larva I), 23.579 ppm (larva II), 25.912 ppm (larva III), 29.155 ppm (larva IV), and 33.877 ppm (pupa). Furthermore, C. clavulatum-synthesized AgNP inhibited dengue (serotype dengue virus type-2 (DEN-2)) viral replication in Vero cells. Notably, 50 μg/ml of green-synthesized AgNP showed no cytotoxicity on Vero cells while reduced DEN-2 viral growth of more than 80%; 12.5 μg/ml inhibited viral growth of more than 50%. Cellular internalization assays highlighted that untreated infected cells showed high intensity of fluorescence emission, which denotes high level of viral internalization. Conversely, AgNP-treated infected cells showed reduced levels of fluorescence, failing to show significant viral load. Overall, our study showed that alga-mediated synthesis of metal nanoparticles may be considered to develop newer, safer, and cheap tools in the fight against the dengue virus, serotype DEN-2, and its vector A. aegypti, with little cytotoxicity on mammalian cells.
登革热是一种由蚊子传播的病毒性疾病，近年来在世界所有地区迅速蔓延。雌性蚊子，主要是埃及伊蚊，传播登革热。在128个国家中，约有39亿人有感染登革热的风险。最近，人们关注了绿色合成纳米颗粒作为Vero细胞中登革热病毒包膜（E）蛋白产生抑制剂和登革热病毒E基因表达下调剂的潜力。藻类是新型化合物的重要来源，可能有助于对抗蚊媒疾病。在本研究中，使用藻类棒形鹿角菜的廉价提取物快速合成了银纳米颗粒（AgNP）。通过紫外可见分光光度法、傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）、能量色散X射线光谱（EDX）和X射线衍射（XRD）对AgNP进行了表征。在灭蚊试验中，棒形鹿角菜提取物对埃及伊蚊幼虫和蛹的LC50值分别为269.361 ppm（一龄幼虫）、309.698 ppm（二龄幼虫）、348.325 ppm（三龄幼虫）、387.637 ppm（四龄幼虫）和446.262 ppm（蛹）。棒形鹿角菜提取物在2,2-二苯基-1-苦基肼（DPPH）和一氧化氮（NO）自由基清除试验中也表现出中等程度的抗氧化活性。棒形鹿角菜合成的AgNP对幼虫和蛹的LC50值分别为21.460 ppm（一龄幼虫）、23.579 ppm（二龄幼虫）、25.912 ppm（三龄幼虫）、29.155 ppm（四龄幼虫）和33.877 ppm（蛹）。此外，棒形鹿角菜合成的AgNP抑制了Vero细胞中登革热（血清型登革热病毒2型（DEN-2））的病毒复制。值得注意的是，50μg/ml的绿色合成AgNP对Vero细胞无细胞毒性，同时使DEN-2病毒生长降低80%以上；12.5μg/ml抑制病毒生长超过50%。细胞内化试验表明，未处理的感染细胞显示出高强度的荧光发射，这表明病毒内化水平较高。相反，AgNP处理的感染细胞荧光水平降低，未显示出显著的病毒载量。总体而言，我们的研究表明，藻类介导的金属纳米颗粒合成可被视为开发更新、更安全、更廉价的工具，用于对抗登革热病毒血清型DEN-2及其传播媒介埃及伊蚊，且对哺乳动物细胞几乎没有细胞毒性。
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