Antigenotoxic Potential of Terminalia chebula fruit (myrobalan) Against Cadmium in Allium Test

Prarthna Jafferey; H.S. Rathore

摘要

Allium cepa bulbs were grown in pure tap water (Group I) and in five concentrations (10-1M to 10-5M) alone (Group II) and all five concentration of cadmium chloride as in group II but each contained myrobalan in it at 0.10 mg/ml (Group III). Parameters of study were mean root length, mitotic index, abnormal mitosis and chromosomal aberrations and morphology of root tips. Cadmium chloride exposure significantly inhibited root growth, declined mitotic index, caused abnormal mitosis and aberrations (Group II). In the presence of myrobalan (Group III) cadmium-induced mitodepression, abnormal mitosis and aberrations could be appreciably prevented. No morphological changes in the root tips of any group could be noticed. Probable protective role of myrobalan is discussed. Introduction Cadmium is environmental pollutant which is both genotoxic (1,2,3) and carcinogenic (4) for human beings and occupationally exposed people are at potentially high health risk (5). Reports on antagonistic herbal compounds towards cadmium toxicity are meager. One report showed cytoprotective role of Glycyrrhizae radix extract and its active component liquiritigenin against Cd-Induced cell death (6) and another revealed attenuation of cadmium chloride - induced oxidative stress and genotoxicity by Pluchea lanceolata (7). Recently myrobalan (fruit of Terminalia chebula) a component of reputed ancient Indian herbal formulation “Trifla” meaning three nuts (Terminalia chebula, Terminalia bellirica (Belliric myrobalan) and Emblica officinalis dried nut) could successfully reduce genotoxicity of lead (8) and aluminium (9) in Allium test. Interestingly, cadmium also exerts genotoxic effects in Allium cepa test model 10,11). The aim of present study was to find out whether myrobalan can also antagonize Cd-genotoxicity. Material and Methods Allium cepaEqual sized (1.5 to 2.00 cm) healthy dry brown pink onion bulbs of commercial variety onions (2n=16) were obtained from the local market.Test herbal drugDried young nuts of medicinal plant Terminalia chebula (myrobalan) locally called “Bal Harad” were procured from local herbal medicine shop. Dried nuts were gently baked for few minutes in steel container. This treatment caused swelling of nuts. After cooling the nuts were grinded in electrical mixer to obtain very fine powder. Powdered material was stored for further use in the present experiment.Selection of Dose of myrobalanEarlier studies from this laboratory (12) had revealed that myrobalan at 0.1 mg/ml concentration did not exert any ill effect. Hence this dose was selected for present study.Test Chemical Cadmium chloride monohydrate as CdCl2.H2O, MW 201.32, and purity 99% of Sarabhai India was used. Salt was dissolved in tap water to prepare solutions of different concentrations ranging from M -1 to M -5 . Experimental design was planned as per internationally accepted protocol (13) which consisted of the following steps.(i)The pink brown dry outer scales and some of the brownish bottom plate of each bulb were removed carefully leaving root primordial intact.(ii)For each concentration of test compound i.e. CdCl2, a series of 12 test tubes were arranged in a test tube rack. Five series of the test tubes were filled with the different molar concentrations (M -1 to M -5 ) of solutions of CdCl2 in tap water (Gr II). Twelve tubes were filled with only pure tap water and maintained to provide control (Gr I). 60 tubes were filled with five concentration of cadmium chloride solution as in Gr II but having myrobalan in it at 0.10 mg/ml concentration (Gr III).(iii)Each descaled onion was placed on the top of each tube with root primordial downward in the liquid.(iv)After 24 hours test suspension in (Gr III) and test solutions in (Gr II) and tap water in (Gr I) were changed. Change of liquid was repeated after 48 hours.(v)After 48 hours two onions out of twelve in each series with most poorly growing roots were removed. Same day i.e. after 48 hours distal 2 mm of five roo

机译：洋葱洋葱鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎的鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞茎鳞片mg / ml（第III组）。研究的参数是平均根长，有丝分裂指数，异常有丝分裂和染色体畸变以及根尖的形态。暴露于氯化镉中会显着抑制根的生长，降低有丝分裂指数，导致异常的有丝分裂和畸变（第二组）。在存在myrobalan（第III组）镉诱导的丝裂抑制的情况下，可以明显预防异常的有丝分裂和畸变。没有观察到任何组的根尖的形态变化。讨论了myrobalan的可能的保护作用。简介镉是环境污染物，对人类而言既具有遗传毒性（1,2,3），又具有致癌性（4），职业接触者具有潜在的高健康风险（5）。关于拮抗草药化合物对镉毒性的报道很少。一份报告显示甘草提取物及其活性成分liquiritigenin对Cd诱导的细胞死亡具有细胞保护作用（6），另一份报告则表明氯化镉减弱了杉木（Pluchea lanceolata）诱导的氧化应激和遗传毒性（7）。最近，myrobalan（Terminalia chebula的果实）是古代印度草药配方“ Trifla”的一种成分，意为三种坚果（Terminalia chebula，Terminalia bellirica（Belliric myrobalan）和Emblica officinalis干果）可以成功地降低铅（8）和铝的遗传毒性（ 9）在葱属测试中。有趣的是，镉在葱属洋葱试验模型10,11中也具有遗传毒性作用。本研究的目的是发现myrobalan是否也可以拮抗镉的基因毒性。材料和方法从当地市场购买大小相等（1.5至2.00厘米）的健康干棕粉洋葱（商品号为2n = 16）的健康干燥棕色粉红色洋葱鳞茎。测试草药干燥的药用植物榄仁（Terminal榄）的年轻坚果在当地称为“ Bal Harad”是从当地草药店购买的。将干燥的坚果在钢制容器中轻轻烘烤几分钟。这种处理导致坚果肿胀。冷却后，将螺母在电动混合器中研磨，以获得非常细的粉末。粉末状的材料被保存以供本实验进一步使用。myrobalan剂量的选择来自该实验室的早期研究（12）显示，浓度为0.1 mg / ml的myrobalan不会产生任何不良作用。因此，选择此剂量进行本研究。测试化学氯化镉一水合物为CdCl2.H2O，MW 201.32，纯度为Sarabhai India的99％。将盐溶解在自来水中以制备浓度范围从M -1到M -5的溶液。根据国际公认的方案（13）计划实验设计，该方案包括以下步骤：（i）小心地除去每个球茎的粉红色棕色干燥的外部鳞屑和一些棕褐色的底板，使根原基完整无损。在每种浓度的测试化合物（即CdCl2）中，在试管架中布置了一系列的12个试管。在五个系列的试管中充满不同摩尔浓度（M -1至M -5）的CdCl2在自来水（Gr II）中的溶液。仅向十二个试管中注入纯净的自来水，并进行维护以提供控制（Gr I）。按照Gr II的要求，将60个试管装满5种浓度的氯化镉溶液，但其中的myrobalan浓度为0.10 mg / ml（Gr III）。（iii）将每个去鳞的洋葱放在每支试管的顶部，将根基从头向下放置（iv）24小时后，更换（Gr III）中的测试悬浮液，（Gr II）中的测试溶液和（Gr I）中的自来水。 48小时后重复换液。（v）48小时后，除去根系生长最差的每个系列中十二个中的两个洋葱。同一天，即48小时后，远端2毫米（五个圆环）

Antigenotoxic Potential of Terminalia chebula fruit (myrobalan) Against Cadmium in Allium Test

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