Anbukkarasi Muniyandi, Thomas Philip A, Sheu Joen-Rong, Geraldine Pitchairaj
Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli - 620 024, Tamil Nadu, India.
Department of Ocular Microbiology, Institute of Ophthalmology, Joseph Eye Hospital, Tiruchirappalli - 620 001, Tamil Nadu, India.
Biomed Pharmacother. 2017 Jul;91:467-475. doi: 10.1016/j.biopha.2017.04.079. Epub 2017 May 4.
Silver nanoparticles (AgNPs) have been found useful in biological systems and in medicine since they possess a large surface area to volume ratio, which confers on them several unique properties. In the present study, AgNPs that had been biosynthesized using an ethanolic extract of Tabernaemontana divaricata leaf were evaluated for putative antioxidant potential and efficacy in preventing experimental in-vitro selenite-induced opacification of the ocular lens (cataractogenesis). The antioxidant potential of the AgNPs was evaluated in-vitro by looking for radical-scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydrogen peroxide (HO) free radicals as well as by determining reducing power. The anticataractogenic potential of the AgNPs was evaluated in an in-vitro model of selenite-induced cataractogenesis in five groups of Wistar rat lenses cultured in Dulbecco's modified Eagle's medium (DMEM) for 24h: Group I lenses (negative control) were cultured in DMEM alone; Group II lenses were exposed to sodium selenite alone (100μM); Group III lenses were exposed simultaneously to sodium selenite and the T. divaricata extract (250μg/ml); Group IV lenses were exposed simultaneously to sodium selenite and the biosynthesized AgNPs (125μg/ml); and Group V lenses were exposed to the AgNPs alone. In these lenses, gross morphological changes, as well as activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), and levels of reduced glutathione (GSH) and malondialdehyde (MDA), were determined. In-vitro, the AgNPs (which were spherical in shape with an average diameter ranging from 15 to 50nm) showed potent and concentration-dependent radical-scavenging activity on DPPH and HO free radicals as well as reducing power. The gross morphological changes seen in the cultured rat lenses were: all eight control (Group I) lenses remained transparent; dense opacification was noted in all eight selenite-challenged untreated (Group II) lenses; in selenite-challenged, simultaneously T. divaricata extract-treated (Group III) lenses, no opacification occurred in seven of eight (87.5%) lenses and only minimal opacification in one (12.5%) lens; all the eight Group IV (selenite-challenged, simultaneously AgNPs-treated) lenses did not show any opacification; and all the eight Group V lenses (exposed to AgNPs alone) remained as transparent as control lenses. The mean activities of CAT, SOD, GPx and GST, and the mean levels of GSH, were significantly (p<0.05) lower in Group II lenses than those in Groups I, III, IV and V lenses, while the mean MDA level was significantly (p<0.05) higher in Group II lenses than those in Groups I, III, IV and V lenses; oxidative damage possibly occurred in Group II lenses, whereas this appears to have been prevented in Groups III and IV lenses. These observations suggest that the T. divaricata leaf ethanolic extract, and also the AgNPs biosynthesized using the T. divaricata extract, possess effective in-vitro antioxidant activity and the potential to prevent experimental selenite-induced opacification in cultured Wistar rat lenses.
银纳米颗粒(AgNPs)因其具有较大的表面积与体积比而具有多种独特性质,已被发现在生物系统和医学领域中具有应用价值。在本研究中,对使用鸡蛋花(Tabernaemontana divaricata)叶乙醇提取物生物合成的AgNPs的假定抗氧化潜力以及预防实验性体外亚硒酸盐诱导的晶状体混浊(白内障形成)的功效进行了评估。通过检测对1,1-二苯基-2-苦基肼(DPPH)和过氧化氢(HO)自由基的自由基清除活性以及测定还原能力,在体外评估了AgNPs的抗氧化潜力。在五组于杜尔贝科改良 Eagle 培养基(DMEM)中培养24小时的Wistar大鼠晶状体的亚硒酸盐诱导白内障形成的体外模型中,评估了AgNPs的抗白内障潜力:第一组晶状体(阴性对照)仅在DMEM中培养;第二组晶状体仅暴露于亚硒酸钠(100μM);第三组晶状体同时暴露于亚硒酸钠和鸡蛋花叶提取物(250μg/ml);第四组晶状体同时暴露于亚硒酸钠和生物合成的AgNPs(125μg/ml);第五组晶状体仅暴露于AgNPs。在这些晶状体中,测定了总体形态变化以及过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GPx)和谷胱甘肽-S-转移酶(GST)的活性,以及还原型谷胱甘肽(GSH)和丙二醛(MDA)的水平。在体外,AgNPs(呈球形,平均直径范围为15至50nm)对DPPH和HO自由基表现出强大的且浓度依赖性的自由基清除活性以及还原能力。在培养的大鼠晶状体中观察到的总体形态变化为:所有八个对照(第一组)晶状体保持透明;所有八个未处理的亚硒酸盐挑战组(第二组)晶状体出现致密混浊;在亚硒酸盐挑战且同时用鸡蛋花叶提取物处理的组(第三组)晶状体中,八个中有七个(87.5%)未出现混浊,仅一个(12.5%)晶状体有轻微混浊;所有八个第四组(亚硒酸盐挑战且同时用AgNPs处理)晶状体未出现任何混浊;所有八个第五组晶状体(仅暴露于AgNPs)与对照晶状体一样保持透明。第二组晶状体中CAT、SOD.GPx和GST的平均活性以及GSH的平均水平显著低于(p<0.05)第一、三、四和五组晶状体,而第二组晶状体中MDA的平均水平显著高于(p<0.05)第一、三、四和五组晶状体;第二组晶状体可能发生了氧化损伤,而第三和第四组晶状体似乎预防了这种损伤。这些观察结果表明,鸡蛋花叶乙醇提取物以及使用该提取物生物合成的AgNPs具有有效的体外抗氧化活性以及预防培养的Wistar大鼠晶状体实验性亚硒酸盐诱导混浊的潜力。