低剂量连续辐射引起的小鼠免疫系统的变化

为了评估低剂量X射线连续辐射对BALB/c小鼠健康机体免疫系统的影响， 实验采用X射线全身连续照射BALB/c小鼠， 照射第一天剂量为0.07 Gy， 剂量率0.2 Gy/min， 之后每天照射0.08 Gy， 共照射12 d， 累积剂量1.03 Gy， 照射后24和48 h取血、胸腺和脾脏。 流式细胞仪检测免疫细胞周期和凋亡的变化， 胸腺和脾脏指数用重量法获取。 实验结果表明， 小鼠胸腺细胞的周期在照射后24 h被阻滞在G2/M期； 外周血淋巴和胸腺细胞周期48 h被阻滞在 G0/G1期， 细胞凋亡比例在照射后两个时间点都显著增加； 脾脏淋巴细胞周期24 h被阻滞在 G0/G1期， 48 h被阻滞在 S期， 细胞凋亡比例在24和48 h显著减少； 脾脏指数在照射后48 h显著减少。 故低剂量X射线连续全身照射BALB/c小鼠可激活免疫细胞不同的周期监测点， 引起免疫细胞凋亡比例发生变化， 造成一定的辐射损伤， 且这种影响随着免疫器官的不同而不同。 For estimating the effect of low doses X ray continual irradiation to immunity system of mouse， BALB/c mice were continually irradiated to 1.03 Gy by X rays at a dose rate of 0.2 Gy/min in 13 d. At 24 or 48 h after irradiation， the immunocyte cycle and apoptosis were determined by flow cytometry， and the thymus and spleen weights were measured too. The results showed that the cycle of thymocyte were arrested in G2/M at 24 h， the number of peripheral blood lymphocytes and thymocytes in G0/G1 phase at 48 h was up and the percentage of apoptosis had a significance increase in both of time points； the cycle of spleen lymphocytes was delayed in G0/G1 at 24 h， in S phase at 48 h， the apoptosis had a significance decrase at 24 and 48 h； spleen index declined significantly at 48 h. The results suggested that low doses continual X ray whole body irradiation could activate different cell cycle checkpoints， and result in some changes of apoptosis and some damages to immunocytes. The continual X ray irradiation affects the organs differently， it might provide experiment basis for radioprotection.     

低剂量电离辐射预处理对不同肿瘤细胞周期的影响

实验研究了低剂量γ射线预照射对人肝癌细胞系hepG2和人宫颈癌细胞系HeLa的细胞周期进程的影响.结果显示(1)低剂量(5cGy)辐射后,两种细胞的G2/M期细胞短暂累积;(2)低剂量辐射促进肿瘤细胞的生长;(3)高剂量(3Gy)辐射后,hepG2细胞发生G2期阻滞,HeLa细胞发生S期和G2期阻滞;(4)与单纯高剂量照射相比,低剂量辐射预处理后4h,再给予高剂量辐射,可进一步促进hepG2细胞在G2/M期累积,但是预照射对HeLa细胞的周期进程没有明显影响.因此,低剂量辐射预处理对高剂量诱导的细胞周期阻滞的影响依赖于肿瘤细胞的类型.     

低剂量辐射诱导hepG2细胞克隆存活和细胞周期适应性反应

以低剂量γ射线(0.05 Gy)预照射人肝癌细胞hep G2， 8 h后再用高剂量(3 Gy)照射， 测定了细胞的克隆存活率和细胞周期。 结果表明， 低剂量辐射预处理可诱导hep G2细胞产生克隆存活适应性反应， 并且有助于细胞通过G2/M期阻滞； 低剂量辐射诱导的克隆存活适应性反应与增强的通过细胞周期阻滞的能力之间有一定的相关性。 Human hepatoma cells hep G2 were irradiated with 3 Gy of γ ray 8 hours after primed with 0.05 Gy of γ ray， thereafter，cell survival and cell cycle were determined. The results indicated that both survival adaptive response and the enhanced ability to overcome G2/M arrest could be induced by pre irradiation with low dose of γ ray. It is suggested that there is a certain correlation between the survival adaptive response and the enhanced ability to overcome cell cycle arrest.     

碳离子辐照对人肺癌细胞A549细胞周期进程的影响

选取对数生长期人肺癌细胞A549接受0—6.0 Gy 碳离子照射， 用克隆形成法检测细胞的存活率； 并于照射后12和24 h收集细胞， 用流式细胞术检测细胞周期各时相的细胞百分比， 观察不同剂量碳离子辐照对A549细胞周期进程的影响。 结果显示: 0—6.0 Gy 碳离子照射后细胞存活率显著下降； 照射后12 h细胞发生G0/G1期阻滞， 而照射后24 h， 1.0 Gy 照射组细胞在G0/G1期阻滞， 2.0—6.0 Gy 照射组细胞在G2/M期阻滞。 上述结果表明， 在A549细胞接受碳离子照射后的12 和24 h内， 1.0 Gy 照射可持续激活细胞G1期检查点， 而2.0—6.0 Gy 碳离子照射后其细胞周期进程是随时间变化的。 To investigate the effects of cell cycle progression of A549 cell induced by 12C6+ ion irradiation at different doses, the survival fractions of the A549 cells were determined by colony forming assay; cell cycles were analyzed by FACS at 12 h or 24 h after irradiation. The results showed that the percentage of survival in the A549 cells decreased with irradiation doses. Compared with control group， the percentage of the cells in G0/G1 phase significantly increased at 12 h after irradiation with different doses of 12C6+ ions. However， at 24 h after irradiation the percentage of the cells in G0/G1 phase significantly increased with 1.0 Gy 12C6+ ions， while the cells showed increasing percentage in G2/M phase with 2.0, 4.0 or 6.0 Gy 12C6+ ions. The results suggested that G1 cell cycle checkpoint was activated in 12—24 h after irradiation with 1.0 Gy 12C6+ ions， but after irradiation with 2.0—6.0 Gy 12C6+ ions， the cell cycle progression of the A549 cells changed with time.     

利用拉曼光谱技术研究低剂量X射线辐射对人神经母细胞瘤细胞的影响

低剂量电离辐射引发的生物效应复杂而多样,其研究往往又受到辐射标志物和检测技术手段的限制。将拉曼光谱技术应用于低剂量辐射生物效应研究,利用10 mW,532 nm共聚焦拉曼光谱对经过100,200和500 mGy三种辐射剂量的X射线辐照之后的人神经母细胞瘤细胞进行检测,发现细胞嘌呤核苷酸（722～728和1 572～1 581 cm-1等等）、嘧啶核苷酸（770～785 cm-1等等）等DNA相关的拉曼特征峰受到电离辐射影响而发生变化,说明低剂量X射线辐照造成细胞DNA水平改变。采用流式细胞术对同样条件辐照后培养6 h的人神经母细胞瘤细胞进行细胞周期分析发现,三种剂量的X射线电离辐射均造成细胞在G2期阻滞,同样提示电离辐射引起DNA水平升高。通过划痕实验分析辐照后20 h的细胞迁移能力,结果显示,相较于未接受X射线照射的对照细胞,受到三种剂量电离辐射的人神经母细胞瘤细胞均出现迁移水平下降。研究结果表明,通过拉曼光谱分析发现低剂量X射线电离辐射引起人神经母细胞瘤细胞DNA水平变化,其结果与细胞周期分析和迁移分析的结果相一致,但检测时间大大提前,利用拉曼光谱技术可以实现低剂量辐射损伤等细胞生物学效应的早期发现与监测。     

重离子对人胃癌细胞DNA错配修复基因MSH2表达的影响

采用高传能线密度(LET)重离子辐照人胃癌SGC7901细胞,应用流式细胞技术、蛋白质印迹法(Western blot)及反转录聚合酶链式反应(RT-PCR)观察重离子诱导人胃癌SGC7901细胞周期、凋亡和MSH2表达状况。结果表明:与对照组相比,SGC7901细胞在辐射后72 h G2/M期所占细胞比率(33.26±0.08)和凋亡率(24.16±0.64)均达到峰值,且呈时间依赖性增加;经重离子照射后,DNA错配修复基因MSH2 m RNA和蛋白表达水平在6 h最高。结果提示:重离子在体外诱导SGC7901细胞周期阻滞和凋亡,且具有显著的时间依赖性效应;重离子在一定剂量和时间下,诱导了SGC7901细胞MSH2基因表达。DNA错配修复基因MSH2可能参与了重离子辐照诱导胃癌细胞DNA损伤的修复应答。     

羧甲基-β-1,3-葡聚糖对人肝癌HepG2细胞的放射增敏作用

本研究旨在探讨羧甲基-β-1，3葡聚糖（CMG）对人肝癌HepG2细胞X射线或12C6+离子束辐射敏感性的影响。首先用CCK-8法检测CMG对HepG2细胞的生长抑制情况，得到半数抑制浓度（IC50）为120.6μg/mL。用浓度为0.1×IC50的CMG预处理HepG2细胞24 h，再给予2 Gy X射线或12C6+离子束辐照（CMG+辐照组）；CMG未处理组直接接受2 Gy X射线或12C6+离子束辐照（辐照组）。对比分析辐照组和CMG+辐照组细胞的克隆存活、DNA损伤、凋亡与周期分布、细胞内活性氧（ROS）水平。发现:与X射线辐照组相比，相同剂量的12C6+离子辐照组克隆存活率更小，DNA损伤和周期阻滞更加严重，细胞凋亡率和细胞内ROS水平也更高。与单独X射线或12C6+离子束辐照组相比，CMG+辐照组克隆存活率明显降低，细胞凋亡率随辐照后CMG作用时间的延长而明显增加，CMG使辐照后细胞内ROS维持在一个较高的水平，同时CMG明显加重了单独辐照诱导的DNA损伤和周期阻滞。结果表明，与X射线相比，HepG2细胞对相同剂量的12C6+离子辐射更敏感；CMG可增加HepG2细胞对X射线或12C6+离子辐射的敏感性；CMG可能通过增加受照HepG2细胞内的ROS水平，加剧辐照诱导的DNA损伤，促进辐射诱导细胞凋亡而起到辐射增敏作用。This study aims to investigate the effect of carboxymethy-β-1, 3-glucan (CMG) on the sensitivity of human hepatoma HepG2 cells to X-rays or 12C6+ ions irradiation. First, the inhibitory effect of CMG on the growth of HepG2 cells was detected by CCK-8 assay, and the half maximal inhibitory concentration (IC50) was 120.6 μg/mL. HepG2 cells were pretreated with CMG at a concentration of 0.1×IC50 for 24 h and then irradiated with 2 Gy X-ray or 12C6+ ion beams (CMG + irradiation group). CMG untreated group was directly irradiated by 2 Gy X-rays or 12C6+ ions beam (irradiation group). The clone survival, DNA damage, cell apoptosis, cell cycle distribution, and intracellular reactive oxygen species (ROS) levels in irradiation group and CMG + irradiation group were comparatively analyzed. The results showed that the clone survival rate was lower, DNA damage and cycle arrest were more serious, and the rate of apoptosis and intracellular ROS levels were higher in 12C6+ ions irradiation group than those in the same dose of X-rays irradiation group. Compared with X-rays or 12C6+ ions irradiation group, the clone survival rate of CMG + irradiation group was significantly decreased, and the apoptosis rate significantly increased with the prolongation of CMG treatment post-irradiation; CMG maintained intracellular ROS at a higher level after irradiation, CMG also significantly aggravated radiation-induced DNA damage and cycle arrest. These results indicated that HepG2 cells were more sensitive to 12C6+ ions radiation than those at the same dose of X-rays. CMG increased the sensitivity of HepG2 cells to X-rays or 12C6+ ions irradiation by increasing intracellular ROS level, exacerbating radiation-induced DNA damage and promoting radiation-induced apoptosis in irradiated HepG2 cells.     

两种保护剂对C离子诱导小鼠急性肝损伤的应答

比较了N-乙酰半胱氨酸(NAC)及乙酰左旋肉毒碱(ALCAR)对12C6+离子照射小鼠的损伤效应,并探讨了其可能的作用机制。利用4Gy剂量的12C6+离子束对预先给予NAC(100mg/kg)和ALCAR(100mg/kg)保护的昆明小鼠进行单次全身照射。随后检测肝组织中总抗氧化能力(TAC)、DNA单链断裂和细胞凋亡率。结果显示,与照射对照组相比,提前给予NAC和ALCAR均极显著地增强了肝组织的抗氧化能力(P0.001),减轻了12C6+离子导致的肝组织中DNA断裂(P0.001)和细胞凋亡(P0.001)。此外,还发现ALCAR组抗重离子辐照损伤的能力显著地高于NAC组(P0.05)。实验结果提示了NAC和ALCAR可通过抵御组织内的氧化胁迫,阻止DNA链的断裂和细胞的凋亡,实现对C离子辐照损伤的保护效应。而且ALCAR比NAC可能更适合成为有潜力、有希望的抗C重离子辐射药物。     

人肝癌SMMC-7721细胞对低剂量γ射线辐射超敏感性的研究

研究了人肝癌细胞SMMC-7721在低剂量γ射线照射下超敏感性和增强的辐射抗性响应。选用对数生长期细胞接受0—6 Gy不同剂量的^60Coγ射线的照射。利用流式细胞仪对细胞进行分选计数，并用克隆形成法检测细胞存活率。发现SMMC-7721细胞存在低剂量辐射超敏感性和增强的辐射抗性响应，即在0—0．3Gy之间细胞表现出单位剂量杀伤增强现象，在0．3—1Gy细胞表现一定的辐射抗性，在1Gy以上，细胞的存活符合线性平方模型。     

不同LET辐射致DNA集簇性损伤的研究

利用辐照质粒DNA构象变化的分子模型,以DNA糖苷酶Fpg和AP核酸内切酶EndoIII识别并切割辐射所致DNA碱基损伤,将其转换为DNA断裂损伤,通过电泳分析DNA分子构象变化,研究比较γ射线、质子和7Li离子诱发DNA集簇性损伤。50Gy以上高剂量γ辐射对质粒DNA的损伤主要表现为单链断裂(SSB)和很少比例的双链断裂(DSB),并能产生一定水平的集簇性损伤。相比之下,高能质子束和高LET的7Li离子直接所致DNA的断裂损伤以及所产生的集簇性碱基损伤比γ射线的要严重,质子10Gy照射就可诱发明显的集簇损伤。     

~(12)C~(6+)离子束辐照紫苏干种子当代效应

利用兰州重离子研究装置(HIRFL)提供的12C6+离子束辐照紫苏干种子(辐照剂量为40,80和120Gy,剂量率4Gy/min),探讨了重离子束辐照对紫苏M1代的生物学效应。结果发现,经不同剂量的12C6+离子束辐照后,紫苏种子的发芽率、发芽势、存活率、株高、分枝数、单株产量和千粒重等生物学性状均发生了变化,其中发芽势、单株产量和千粒重随辐照剂量的提高而降低,且有明显的剂量效应关系,但发芽率、大田成活率、株高和分枝数却随辐照剂量的增大,呈现出明显的"抛物线"趋势;紫苏幼苗根尖细胞的微核率和染色体畸变率随辐照剂量增加呈线性增加关系。这表明:12C6+重离子束辐照紫苏种子,具有明显的当代损伤效应,在本试验剂量范围内,低剂量辐照对发芽率和成活率有促进作用。     

12C6+离子束辐照菘蓝干种子当代效应

利用兰州重离子研究装置(HIRFL) 提供的12C6+ 离子束辐照菘蓝干种子(辐照剂量为10,35,60,90 和140 Gy,剂量率20 Gy/min),探讨了重离子束辐照对菘蓝M1代的生物学效应。研究发现,不同剂量的12C6+ 离子束辐照后,菘蓝种子的发芽率、成苗率、株高、根长和根冠比等生物学性状以及对菘蓝中靛玉红和4(3H) 喹唑酮含量均发生了变化,其中株高和根长随辐照剂量的增加而降低;菘蓝叶和根中的4(3H) 喹唑酮和靛玉红的含量随辐照剂量增加呈马鞍形增加关系。这表明:12C6+ 重离子束辐照菘蓝种子具有明显的当代损伤效应, 并可显著提高菘蓝中靛玉红和4(3H) 喹唑酮的含量,其辐照适宜诱变剂量为35 Gy。To investigate the M1 biological effects of heavy ions on Isatis indigotica Fort, its dry seeds were irradiated by 12C6+ beam with the dose of 0, 10, 35, 60, 90 and 140 Gy respectively,at the rate of 20 Gy/min delivered by the Heavy Ion Research Facility in Lanzhou (HIRFL). The results showed that biological characters such as germinating rate, germinating potential, survival rate, plant height, root height and root-shoot ratio were changed after irradiation. Moreover, the plant height and root height decreased in a dos dependent manner. The indirubin and 4(3H) quinazolinone content of Isatis indigotica Fort was improved and exhibited obviously “saddle” trends with irradiation dose increasing.Data suggest that exposure with low-dose 12C6+ to seeds of Isatis indigotica Fort has obvious injury effects at the first generation, and the active ingredient content of Isatis indigotica Fort may be improved by carbon ion beamirradiation. It is concluded that the suitable irradiation dose of mutation breeding is 35 Gy for the seeds of Isatis indigotica Fort.     

姜黄素对重离子辐射损伤小鼠睾丸组织的防护作用研究

探讨姜黄素(Curcumin,简称Cur)对重离子辐射损伤小鼠睾丸组织的防护作用。小鼠灌胃不同剂量的Cur后给予4 Gy剂量~(12)C~(6+)离子束全身单次照射。24 h后对小鼠睾丸组织形态学变化进行观察,并测定丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性和过氧化氢酶(CAT)活性。结果显示:中、高浓度Cur预处理组对小鼠睾丸组织的形态具有较好的保护作用;低、中浓度Cur预处理组MDA含量显著降低(P0.05);与单纯照射组相比,低浓度Cur预处理组SOD活性水平和中浓度Cur预处理组CA了活性水平显著提高(P0.05)。结果表明:Cur对重离子全身辐射小鼠的抗氧化系统有一定的激活效应,对辐射损伤有一定的防护作用,其机制可能与Cur清除自由基,保护脂质和蛋白质有关。     

12C6+离子对阿维链霉菌累进辐照诱变效应研究

选用12C6+ 离子束对阿维链霉菌诱变选育高产菌株与原始菌株进行辐照诱变, 研究其累进辐照效应。实验结果表明,在辐照剂量为10 Gy时, 原始菌株比诱变高产菌株存活率高, 抗辐射能力强;辐照剂量高于30 Gy时,诱变高产菌株比原始菌株存活率高, 抗辐射能力强。原始菌株正突变率最高的辐照剂量为50 Gy, 致死率99.43%,正突变率最高, 达34.2%;对诱变高产菌株辐照剂量为30 Gy,致死率94.97%,正突变率最高, 达23.5% 。累进辐照效应降低了最佳辐照剂量。 Mutagenic effect on the mutant high producing strain ZJAV Y1 203 and the original strain ZJAV A1 irradiated by ion beam of 12C6+ have been investigated. The experimental results indicated that the original strain has higher survival rate and stronger resistance to radiation than mutant high producing Strain at dose of 10 Gy. The mutant high producing strain has higher survival rate and stronger resistance to radiation than the original strain at the dose higher than 30 Gy. The lethality was 97% and the highest rate of orthomutation was 34.2%, when ZJAV A1 was irradiated by 50 Gy 12C6+ beam. The lethality was 94.97% and the highest rate of orthomutation was 23.5% when ZJAV Y1 203 was irradiated by 30 Gy 12C6+ beam. The best radiation dose is decreased by progressivity irradiation.     

12C6+束辐照引起的实验猪皮肤急性损伤反应的研究

探讨了正常皮肤对重离子辐照急性损伤反应的耐受性, 为重离子治癌临床应用提供安全性检测的实验依据。实验前10 min, 实验猪肌肉注射复方氯胺酮1.2 mg/kg进行麻醉, 然后在兰州重离子研究装置（HIRFL）辐照终端利用12C6+束照射, 辐照剂量分别为0, 12, 21和27 Gy, 辐照分3次完成, 剂量率约为1.2 Gy/min, Bragg峰区照射, 辐照后每隔7 d对照射野拍照并活检取样, 做HE组织病理学观察。不同剂量12C6+离子束辐照实验猪皮肤后, 皮肤外观反应随辐照剂量增大而加快, 表现为肿胀和色素沉积等; 皮肤组织结构的变化明显, 上皮细胞排列紊乱、 萎缩、 空泡变性; 基本恢复正常所需时间也越长, 且都存在明显的剂量效应关系。结果表明, 辐照剂量范围为0—27 Gy时, 重离子对正常皮肤的辐照是安全的。The tolerance of the normal skin to the acute radiation injury reaction induced by heavy ion beams has been studied experimentally. The experimental pigs were injected with 1.2 mg/kg ketamine in 10 min before irradiation and were irradiated with 0, 12 , 21 and 27 Gy 12C6+ ion at a dose rate of 1.2 Gy/min at the Heavy Ion Research Facility in Lanzhou（HIRFL）. The total radiation dose was finished by 3 times at Bragg Peak Region of Heavy Ion Beams.The radiation fields of skin were taken photo and performed biopsy. The contaneous tissues of radiation fields were stained by HE and examined histopatholo gical changes every seven days after irradiation. The results indicated that the cutaneous appearance reaction became more faster with radiation dosage rising and presented with swollen, melanin forming and so on after irradiated by the carbon ions at different dosage. The Pathological examination showed noticeable changes in histological and structural of experimental pigs skin, such as atrophy, vacuolation, denaturation and arranged irregularly in epithelial cells. Furthermore, the time for return to normality became longer with the increasing of radiation dosage. All indexes demonstrated correlation between the does and effects. It is concluded that the irradiation of heavy ion beams to normal skin is security when the radiation dose range is about 0—27 Gy.     

12C6+离子束辐照番茄种子当代效应

利用辐射能量为80 MeV/u 的12C6+重离子束辐照番茄种子, 辐照剂量分别为30, 60, 90, 120和160 Gy研究其对番茄M1代的生物学效应。结果表明, 随着辐照剂量的增大, 番茄的发芽率和成苗率降低, 且成苗率明显低于发芽率, 发现辐照损伤主要抑制了根的生长; MDA和脯氨酸含量变化的总体趋势为随着辐照剂量的增大先升后降再升高, 说明高剂量C离子辐照对生物膜造成更严重的损伤; APX活性随着辐照剂量的增大呈先升后降再升高的趋势, 表明APX在清除活性氧中起主要作用; POD和SOD活性的总体趋势是随着辐照剂量的增大而降低, 且明显低于对照组。综合分析表明, 12C6+重离子束辐照番茄种子, 对M1代具有明显的损伤效应, 高剂量辐照对番茄种子造成的损伤更大, 使酶的活性降低, 抑制植株生长。 To investigate the M1 biological effects of heavy ions irradiation on Lycopersicon esculentum Mill., its seeds were irradiated by 12C6+heavy ions (80 MeV/u) with the dosages of 30, 60, 90, 120 and 160 Gy respectively . The results showed that with doses increased gradually, germination rate and seedling rate of Lycopersicon esculentum Mill. were decreased, and the latter was lower than the former, mainly due to the inhibition of root growth. The irradiation increased the content of MDA and proline evidently, showing irradiation could damage biomembrane, and also decreased the activities of POD and SOD with distinct inhibition pattern. However, the low dose and high dose irradiation promoted APX activity, illustrating APX was induced to protect irradiation injury. In brief, exposure to 12C6+ heavy ions had obvious injury effects on the seeds of Lycopersicon esculentum Mill.. Heavy ions irradiation damaged biomembrane, inhibited activities of enzymes, and finally inhibited the growth of the first generation of these seeds.