Preclinical studies on Orientin, as a countermeasure against radiation injury

Effects of radiation and Orientin in expression genes in bone marrow. Radiation Treatment:C57BL/6 male mice (8-10 weeks old) were divided into four groups: Irradiated control (IR); Orientin+Irradiated (ORI+IR);Vehicle treated and unirradiated (CT) mice and Orientin alone treated mice (ORI). Unanesthesized mice were exposed to 6Gy TBI (gamma rays) at a dose rate of 0.95Gy/min. Orientin was administered subcutaneously at 5mg/kg body weight in 150µl volume/mouse 24h and 3h pre irradiation while untreated control animals received PBS alone. Gene expression: An amplification-free, hybridization based direct digital counting method developed by NanoString Technologies was used for gene expression analysis using mouse cancer immune panel (NS_Mm_CancerImm_C3400). RNA was extracted from bone marrow cells on day 9 after irradiation using Ambion miRVANA kit and a digital multiplexed Nanostring nCounter gene expression assay was performe. Raw data from the NanoString assays were technically normalized followed by biological normalization with a set of 20 housekeeping genes.

辐射与荭草素（Orientin）对骨髓基因表达的影响。辐射处理：将8~10周龄的C57BL/6雄性小鼠分为四组：照射对照组（IR）、荭草素联合照射组（ORI+IR）、溶剂处理未照射对照组（CT）以及单独荭草素处理组（ORI）。对未麻醉小鼠以0.95Gy/min的剂量率实施6Gy全身照射（TBI，伽马射线）。荭草素以5mg/kg体重的剂量、每只小鼠150µl的给药体积，于照射前24小时和3小时皮下注射；未处理对照组仅给予磷酸盐缓冲液（PBS）。基因表达分析：采用由NanoString Technologies（纳米String技术公司）开发的无需扩增、基于杂交的直接数字计数方法，搭配小鼠癌症免疫组试剂盒（NS_Mm_CancerImm_C3400）开展基因表达分析。于照射后第9天从骨髓细胞中提取RNA，使用Ambion miRVANA试剂盒，并进行数字多重Nanostring nCounter基因表达检测。针对NanoString检测得到的原始数据，先进行技术标准化处理，随后以包含20个持家基因的数据集完成生物学标准化。