重离子辐射肺脏的肺肾旁效应及益气滋阴法防护机制研究

The radiation biology pay more and more attention to the mechanism and protection method of the heavy ion induced bystander effects because heavy ion radiation could induce more serious damage to adjacent cells and adjacent organs than derect radiation sites. Heavy ion beam is served as the best anticancer way in the 21th centry, however, the mechanism and protection method of bystander effects should be understanded deeply.This thesis is based on the Related theory of lung and kidney and the storage essence theory of kidney. After the 12C6+ ion beam irradiation to the the rat's upper part of the lungs; Using biological technology to detect lung (containing the adjacent tissues) renal epithelial cells, bone marrow cells after injury repair, cell proliferation and apoptosis, DNA double strand breaks and repair capacity etc.; Using immunohistochemistry and biochip technology to detect ACTH, Cor, E2 etc.related to the adrenal gland axis, hypothalamus pituitary gonadal axis and to detect the change of related factor of the renin angiotensin system; Using methylation sensitive amplified polymorphism (MSAP) to detect the changes of the lung's renal epithelial cells and the changes of bone marrow whole genome methylation, Through the above analysis of the results to analyse side effect of carbon ion radiation damage to the lung tissues and the adjacent organs and its mechanism; According to the TCM theory of mutual promotion between the lung and kidney and by using the method of benefiting qi and nourishing yin to intervene and treat with the compound of traditional Chinese drugs''huangqitang",and detect the changes of these indicators, aim to provide scientific basis for the protection of heavy ion bystander damage.

重离子被誉为21世纪放疗最理想射线，但其对旁细胞及器官可引起辐射旁效应，甚至可能比受辐射部位的损伤更严重，其机制及有效防护亟待系统研究。本课题基于肺肾相关及肾藏精理论，通过碳离子束辐射大鼠肺上部后，利用现代生物学技术检测肺（含旁组织）、肾上皮、骨髓等组织细胞增殖与凋亡、染色体异型、损伤后修复情况及肺P物质；免疫化学、生物芯片等技术检测血中肾上腺轴、下丘脑-垂体-性腺轴、肾素-血管紧张素系统相关因子及EPO、ROS、NO等含量的变化；甲基化敏感扩增多态性(MSAP)法检测肺、肾上皮、骨髓细胞基因组甲基化情况，以期探讨重离子辐射对肺旁组织及旁器官（重点为肾）的旁效应及其机制；并用益气滋阴方黄芪汤对模型大鼠进行干预，检测其对上述指标的影响，为中医药防护重离子辐射旁效应损伤提供科学依据。

放射治疗是治疗恶性肿瘤的重要手段之一，重离子束是目前最理想的放疗射线。但是在临床上用重离子束照射肿瘤部位时，可导致机体未照射部位的组织或器官产生辐射旁效应，包括DNA损伤、表观遗传学改变、基因表达改变、细胞增殖和凋亡、炎症反应等。. 本课题用重离子束照射大鼠右肺部，铅皮屏蔽身体其余部位建立辐射损伤模型，以细胞凋亡、DNA损伤、炎症反应、全基因组甲基化改变、氧化应激及对内分泌网络系统的影响为检测终点，在照射后的不同时间点检测右肺及未受照射组织左肺、左肾及心、肝、脾、骨髓等的变化，并且预先用归芪益元膏干预，探讨其防护重离子辐射旁效应损伤的作用机制，为中医药防护辐射旁效应损伤提供新思路、新方法。. 研究结果发现，重离子束照射大鼠右肺部后，左肺、左肾组织产生了与直接照射右肺相似的细胞凋亡、DNA损伤、炎症反应，肾脏中血管紧张素Ⅱ、血管紧张素Ⅰ型受体表达明显升高，股骨骨髓细胞周期出现明显变化，外周血象明显下降，左肺、左肾、心、肝、脾组织全基因组低甲基化表达及氧化应激反应。表明右肺受照后，左肺、心、肝、脾、肾也对辐射产生了响应，辐射诱导了体内旁效应的发生。预先灌胃归芪益元膏后对上述损伤皆有所改善，表明归芪益元膏对重离子辐射诱发的体内旁效应有一定的防护作用。