珍珠菜提取物ZE4对宫颈癌抑制作用的初步研究

目的观察珍珠菜提取部位ZE4对宫颈癌体内外的抑制作用。方法体外采用MTT法、3H-TdR参入法、集落形成实验观察药物对HeLa细胞的杀伤作用,Hoechst荧光染色法观察药物对HeLa细胞凋亡的诱导作用,划痕实验检测药物对HeLa细胞迁移的影响。体内以抑瘤率和脏器指数为指标观察药物对荷U14小鼠的作用。结果ZE4可抑制He-La细胞的增殖和迁移,并可诱导细胞发生凋亡,3H-TdR实验结果显示药物作用48h的IC50为40.56mg·L-1;对荷U14小鼠给予400mg·kg-1药物其抑瘤率可达49.9%。结论ZE4对宫颈癌具有很好的抑制作用。     

五味沙棘散提高电离辐射后小鼠存活率及保护细胞增殖的效果观察

目的探索验证和开发新型辐射防护药物五味沙棘散的电子辐射防护效果。方法综合采用小鼠30 d活存率实验及细胞增殖实验、流式细胞术检测细胞周期实验,阐明五味沙棘散的辐射防护作用机制。结果五味沙棘散组明显优于模型组,其中高剂量组可以显著延长辐射后死亡小鼠平均存活天数。五味沙棘散5μg/ml可以明显提高辐射后V79细胞增殖能力,与4 Gy组比较,细胞吸光度分别从0.59、0.86、1.17提高到0.69、1.08、1.95,差异非常显著(P<0.01)。五味沙棘散可以通过缓解G2期阻滞,增加G1期比例,保护V79细胞免于辐射诱导的DNA损伤发挥辐射防护作用。结论蒙药五味沙棘散具有很好的电离辐射防护作用。     

4’，5，7三羟基异黄酮抑制骨髓瘤基质细胞bcl-2，bcl-xl，CyclinD1，ICAM-1和IL-6基因表达

目的:研究4’,5,7三羟基异黄酮(Genistein)对人多发骨髓瘤(MM)基质细胞增殖和bcl-2,bcl-xl,CyclinD1,ICAM-1和IL-6基因表达的影响。方法:体外培养人多发骨髓瘤基质细胞,依据浓度梯度分别给予Genistein,用噻唑蓝染色法(MTT)观察Genistein对骨髓瘤基质细胞增殖的影响;10,20,40 mg.L-1Genistein与溶剂对照组分别作用骨髓瘤基质细胞48 h后,半定量RT-PCR法检测骨髓瘤基质细胞bcl-2,bcl-xl,CyclinD1,ICAM-1和IL-6基因表达。结果:Genistein作用骨髓瘤基质细胞48 h,随浓度增加,细胞增殖逐渐下降;10,20,40 mg.L-1Genistein处理组bcl-2,bcl-xl,CyclinD1,ICAM-1,IL-6基因mRNA的表达呈浓度依赖性下降,与溶剂对照组比较,差异有统计学意义(P<0.05,P<0.01)。结论:Genistein可能通过下调骨髓瘤基质细胞bcl-2,bcl-xl,CyclinD1,ICAM-1和IL-6基因mRNA的表达,抑制骨髓瘤基质细胞的生长。     

4′,5′,7-三羟基异黄酮对NB4人急性早幼粒白血病细胞生长和分化的影响

目的：研究4′,5′,7-三羟基异黄酮（genistein）对NB4白血病细胞生长和分化的影响,揭示其诱导细胞凋亡作用。方法：分别用NBT还原力分析、DNA电泳分析观察药物诱导分化和凋亡作用;Northern杂交分析和质粒DNA断裂分析检测bcl-2基因表达和DNA拓扑异构酶II活性。结果：Genistein可明显抑制NB4细胞生长,并致细胞大量死亡。在浓度高于40 μmol.L^-1时,可诱导典型的凋亡形态,同时,细胞bcl-2基因表达受到抑制。Genistein可促进DNA拓扑异构酶II介导的DNA断裂, 但诱导NB4细胞分化作用较弱。结论：Genistein通过诱导细胞凋亡来杀伤和抑制NB4细胞的生长,其机制与抑制DNA拓扑异构酶II活性有关。     

雌二醇通过雌激素受体α/糖原合酶激酶-3β/β联蛋白信号通路调节成骨细胞增殖

目的研究观察雌二醇对小鼠成骨细胞前体细胞增殖影响并研究经典wnt通路在其增殖过程中的作用。方法本研究利用小鼠胚胎成骨细胞前体细胞Mc3T3-E1为细胞模型,不同浓度雌二醇处理Mc3T3-E1细胞,四甲基偶氮唑蓝(MTT)检测细胞增殖率,蛋白印迹法和免疫荧光技术检测经典Wnt信号通路信号分子表达。结果研究表明不同浓度雌二醇处理Mc3T3-E1细胞,MTT显示随着雌二醇浓度增加,Mc3T3-E1细胞增殖率增加,其中以10-7 mol/L浓度处理后细胞获得最佳增殖率。蛋白印迹法结果显示随着雌激素浓度增加,小鼠Mc3T3-E1细胞中p-GSK-3β,β联蛋白(catenin),细胞周期蛋白(Cyclin)D1的表达增加。经雌激素受体(ER)α沉默后,相对于空白对照组小鼠Mc3T3-E1细胞p-GSK-3β,β-catenin及Cyclin D1表达下降,而ERβ沉默后,同对照组相比成骨细胞中各分子表达则无明显变化。结论雌二醇可通过ERα/GSK-3β/β-catenin信号通路调节小鼠成骨细胞增殖。     

红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用

目的探讨红三叶异黄酮对人宫颈癌HeLa细胞的抑制作用及其作用机制。方法以MTT法检测红三叶异黄酮20、40、80μg/m L分别在24、48、72 h对人宫颈癌HeLa细胞增殖作用的影响,并在倒置显微镜下观察HeLa细胞形态学改变。流式细胞仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡的影响。实时荧光定量PCR仪检测红三叶异黄酮20、40、80μg/m L对HeLa细胞凋亡中调节基因Bax、Bcl-2表达水平的影响。结果红三叶异黄酮对人宫颈癌HeLa细胞的增殖抑制作用,表现为时间、浓度的相关性。红三叶异黄酮对HeLa细胞的形态学改变,表现为随药物浓度增加细胞固缩变小,并出现凋亡小体。流式细胞仪检测分析显示红三叶异黄酮能引起HeLa细胞凋亡,并随药物浓度的增加而明显,当红三叶异黄酮达到一定有效浓度时,诱导其细胞凋亡。通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖,能使Bax表达上升、Bcl-2表达下降,且呈剂量相关性。结论红三叶异黄酮对人宫颈癌HeLa细胞的增殖具有抑制作用,并呈一定的剂量和时间相关性,提示通过调节Bax mRNA与Bcl-2 mRNA的表达,促进抑制HeLa细胞增殖。     

大豆异黄酮活性组分对H2O2诱导的PC12细胞损伤的影响

目的:探讨大豆异黄酮活性组分对H2 O2 诱导的PC12细胞损伤的保护作用。方法:以H2 O2 损伤PC12细胞为氧化应激损伤的模型,采用光学显微镜观察细胞形态,MTT法判断细胞的存活率,LDH法检测细胞的损伤程度。结果:不同浓度大豆异黄酮能明显改善H2 O2 导致的细胞甲瓒减少,大豆异黄酮处理组细胞存活率明显高于H2 O2 处理组(P <0 .0 1) ;而LDH漏出率则明显低于H2 O2 处理组(P <0 .0 5 )。结论:大豆异黄酮活性组分对H2 O2 诱导的PC12细胞损伤具有保护作用。     

9401号药对小鼠皮肤放射反应试验

研究目的：烟酰胺是近年来国内外研究较多的放射增敏剂,经研究发现,它对体外培养的恶性肿瘤细胞以及对人类恶性肿瘤移植瘤均有明显的放疗增敏作用,而且毒性低。9401号是以烟酰胺为母体化合物合成的烟酰氨基酸类化合物,其放射增敏作用比母体化合物烟酰胺效果更好,而且毒性更低。为了观察9401号对正常组织是否有增敏作用,我们研究了9401号对小鼠皮肤的放射增敏作用。材料与方法：实验动物：选用ICR小鼠,由中国医学科学院实验动物研究所提供,体重18—22g。许可证号：SCXK京2004—0001。药物：9401号（N-烟酰基-亚氨基二乙酸）,为白色粉状晶体,含量大于99％,熔点182～191℃。由中国医学科学院放射医学研究所药物室合成。照射方法：照射采用。Coγ射线垂直照射,剂量率81．79cGy／min。一个剂量点一次照射4只小鼠,将小鼠放入特制的有机玻璃模型内固定,仅照射小鼠后腿,其它部位用铅快保护。采用Denekamp9单次照射法。实验动物分组：实验分为单纯照射组和加药照射组。动物给药剂量为1000mg／kg体重（半数致死量的四分之一）,药物浓度为100mg／ml,于照射前2-3小时一次腹腔给药。照射剂量分别为1Gy,2Gy,3Gy,4Gy,5Gy和6Gy,每-剂量组8只小鼠。     

牛至挥发油体内外抗痢疾杆菌的作用

目的观察牛至挥发油对痢疾杆菌的抑菌和杀菌作用。方法应用宋内痢疾杆菌和福氏痢疾杆菌F2a腹腔感染小鼠，观察牛至挥发油不同时间灌胃给药后对染菌小鼠的保护作用；体外测定该挥发油对痢疾杆菌不同菌群的最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。结果牛至挥发油对感染宋内痢疾杆菌、福氏痢疾杆菌F2a的小鼠有保护作用，对痢疾杆菌不同菌群有明显的抑菌和杀菌作用，MIC和MBC分别为50～100 和125～500 mg · mL 1。结论牛至挥发油是一种抗痢疾杆菌感染的有效药物。     

卷柏中单糖抗辐射作用的研究

目的：分析卷柏（Selaginella tamariscina）中糖类化学成分,观察其对中国仓鼠肺成纤维细胞（V79）和BALB/c小鼠的辐射防护作用,为探讨卷柏抗辐射作用机制提供参考依据。方法：薄层色谱法分析卷柏中糖类化学成分;V79细胞经木糖、果糖、阿拉伯糖处理24 h后接受60Co γ射线照射,分别观察细胞活力、克隆形成率和细胞周期;BALB/c小鼠接受6 Gy 60Co γ射线照射前1周给予木糖,观察受照射小鼠的 30 d存活率。结果：卷柏中含有葡萄糖、木糖、果糖、阿拉伯糖、海藻糖等;其中木糖、果糖、阿拉伯糖在20~200 μg?mL-1浓度范围内可剂量依赖地促进照射后V79细胞增殖,在200 μg?mL-1时可提高照射后V79细胞克隆形成率,增加S期细胞比例,缓解G2/M期阻滞,修正紊乱的细胞周期;照射前分别给予木糖100,500 mg?kg-1可剂量依赖地提高受照射小鼠存活率达20%,30%。结论：卷柏中的单糖类成分可提高受照射小鼠的30 d存活率,其机制可能是通过调节受照射后紊乱的细胞周期而发挥辐射防护作用。     

In vitro and in vivo protective effects of granulocyte colony-stimulating factor against radiation-induced intestinal injury

Intestinal injury is a major cause of death after high-dose radiation exposure. The use of granulocyte-colony stimulating factor (G-CSF) to treat radiation injury has focused on enhancing recovery from hematopoietic radiation syndrome. We evaluated G-CSF for its ability to protect against radiation-induced intestinal injury in rat intestinal epithelial cells (IEC-6) and BALB/c mouse models. For in vitro tests, pre-radiation addition of G-CSF to IEC-6 prevented cytotoxicity and the loss of cell viability. Pre-radiation G-CSF treatment also reduced radiation-induced cleavage of caspase-3 and p53 in IEC-6. For in vivo tests, examination 12 h after abdominal irradiation showed that G-CSF-treated mice were protected against apoptosis of the jejunal crypts. G-CSF-treated mice also showed attenuated intestinal morphological changes 3.5 days after abdominal radiation (10 Gy). G-CSF also reduced the levels of proinflammatory cytokines interleukin-6 and tumor necrosis factor-α after radiation. This study showed that G-CSF may protect against radiation-induced intestinal damage through its anti-apoptotic and anti-inflammatory effects. These results suggest that G-CSF is promising candidate for protection against intestinal mucosal injury following irradiation.     

Effects of radioprotective compounds and anti-inflammatory agents on the acute toxicity of trichothecenes

Antidotic effects of radioprotective and anti-inflammatory drugs against the acute toxicity of T-2 toxin and fusarenon-X, the trichothecene mycotoxins of Fusarium species, were examined in mice and rats with two toxicological parameters, the lethal toxicity and the increment of intestinal fluid volume. The s.c. and i.v. LD50 values of T-2 in male 6-week-old mice were 2.1 and 3.8 mg/kg, respectively, and the s.c. LD50 in 4-week-old mice was 1.6 mg/kg. Pretreatment of mice with steroidal anti-inflammatory agents such as prednisolone and hydrocortisone resulted in a marked reduction in the lethal toxicity of T-2 toxin. However, pretreatment of mice with several radioprotective agents exhibited neither a prolongation of survival time nor a decrease in lethal toxicity in the mice. Enteropooling assay revealed that the steroidal anti-inflammatory drugs also depressed the trichothecene-induced increment of intestinal fluid volume in rats, while no significant effects were observed with non-steroidal inhibitors of the arachidonate cascade. In rats given fusarenon-X, the content of cyclic AMP in the mucosa of ileum was dose- and time-dependently increased, but its maximal increment was only 1.5-fold of the control value. These findings indicate an involvement of the action site of steroidal anti-inflammatory drugs in the development of acute toxicity of trichothecenes in rodents.     

Experimental exposure of arsenic in cultured rat intestinal epithelial cells and cell line: toxicological consequences.

Arsenic is a naturally occurring metalloid and the drinking water contamination by inorganic arsenic remains a major public health problem. The trivalent arsenic (arsenite) is more toxic than the pentavalent form (arsenate), and is known to cause gastrointestinal toxicity. Specific immortal cell lines are considered to be suitable for toxicity screening and testing of chemicals as they are easy to handle and possess most of the biochemical pathways present in the corresponding cells present in vivo. The present study was designed to evaluate and compare the in vitro toxicity of arsenite on rat intestinal epithelial cell line (IEC-6) and primary cultures of rat intestinal epithelial cells (IEC). To evaluate in vitro toxicity, cultures of IEC and IEC-6 cells were assessed for viability, morphometric analysis, membrane transport enzymes and structural constituents for membrane damage, dehydrogenase activity test for respiratory and energy producing processes and esterase activity test for intra and extra cellular degradation, following the post exposures to arsenite (0-20 ppm). Significantly similar concentration-dependent changes in these toxicity-screening parameters in IEC and IEC-6 were observed. Highest tested concentration of arsenite (20 ppm) was found to be detrimental in both IEC and IEC-6. Furthermore, to evaluate arsenite toxicity in epithelial cells of rat intestine, intestinal loops were filled with arsenite solutions and incubated for 30 min in situ. In situ studies also showed a significant arsenite concentration-dependent decline in epithelial cell membrane transport enzyme activities and total hexose and sialic acid contents. Concomitant release of membrane enzymes, hexose and sialic acid in the intestinal luminal fluid following higher arsenite exposures further indicated partial membrane damage. Similar morphological changes in IEC and IEC-6 were also evident. These findings also suggest that IEC-6 cell lines are suitable for initial screening of gastrointestinal cellular toxicity caused by arsenite.     

Ginsenoside Rd prevents and rescues rat intestinal epithelial cells from irradiation-induced apoptosis

Panax ginseng has been shown to have a protective effect for irradiated animals or cells. Ginsenosides are the most active components isolated from ginseng, and ginsenoside Rd has been identified as one of the effective compounds responsible for the pharmaceutical actions of ginseng. In the present study, we studied the molecular mechanisms for the radio-protective action of ginsenoside Rd in rat intestinal epithelial IEC-6 cells. Cells were irradiated with gamma-ray, and apoptosis was examined using Hoechst staining and Western blot analysis. Treatment with ginsenoside Rd before gamma-irradiation inhibited irradiation-induced apoptosis in IEC-6 cells. Administration of Rd after irradiation also inhibited apoptosis in these cells. Irradiation of IEC-6 cells resulted in inactivation of Akt phosphorylation that was abrogated by Rd. On the other hand, irradiation activated phosphorylation of ERK1/2 but did not affect that of p38 MAPK. Inhibition of Akt phosphorylation prevented the reduction of apoptosis by Rd following irradiation. Pretreatment with an inhibitor of the MEK pathway further decreased the number of apoptotic cells. Rd decreased the ratios of Bax/Bcl-2 and Bax/Bcl-xL, the levels of cytochrome c, and the cleaved form of caspase-3 in irradiated IEC-6 cells. Our results suggest that ginsenoside Rd protects and rescues rat intestinal epithelial cells from irradiation-induced apoptosis through a pathway requiring activation of PI3K/Akt, inactivation of MEK, and also inhibition of a mitochondria/caspase pathway.     

Hydrogen attenuates radiation-induced intestinal damage by reducing oxidative stress and inflammatory response

The small intestine is known to be particularly sensitive to radiation, and the major limiting factor of radiotherapy is the gastrointestinal syndrome that subsequently develops after its administration. The detrimental effects of radiation are mostly mediated via the overproduction of reactive oxygen species (ROS), especially the hydroxyl radical (·OH). Because hydrogen is a selective ·OH scavenger, we hypothesized that hydrogen might exert a protective effect against radiation-induced intestinal damage. Herein, radiation models were built both in mice and in an intestinal crypt epithelial cell (IEC-6) line. In the animal experiment, we demonstrated that hydrogen-rich saline significantly reduced radiation-induced intestinal mucosal damage, improved intestinal function, and increased the survival rate. In addition, radiation-induced oxidative stress damage and systemic inflammatory response were also mitigated by hydrogen treatment. Moreover, hydrogen treatment decreased cell apoptosis and maintained intestinal epithelial cell proliferation in mice. In vitro experiments using the IEC-6 cell line showed that hydrogen-rich medium significantly inhibited ROS formation, maintained cell viability, and inhibited cell apoptosis. Importantly, hydrogen treatment prevented mitochondrial depolarization, cytochrome c release, and activity of caspase-3, caspase-9, and PARP. Moreover, the decreased expression of Bcl-xl and Bcl-2 and the increased expression of Bax protein were also blocked by hydrogen treatment. In conclusion, our study concurrently demonstrated that hydrogen provides an obviously protective effect on radiation-induced intestinal and cell injuries. Our work demonstrated that this protective effect might be due to the blockage of the mitochondrial apoptotic pathway.     

Geraniin down regulates gamma radiation-induced apoptosis by suppressing DNA damage

Gamma ray irradiation triggers DNA damage and apoptosis of proliferating stem cells and peripheral immune cells, resulting in the destruction of intestinal crypts and lymphoid system. Geraniin is a natural compound extracts from an aquatic plant Nymphaea tetragona and possesses good antioxidant property. In this study, we demonstrate that geraniin rescues radiosensitive splenocytes and jejunal crypt cells from radiation-induced DNA damage and apoptosis. Isolated splenocytes from C57BL/6 mice treated with geraniin were protected against radiation injury of 2 Gy irradiation through the enhancement of the proliferation and attenuation of DNA damage. Also, geraniin inhibited apoptosis in radiosensitive splenocytes by reducing the expression level and immunoreactivity of proapoptotic p53 and Bax and increasing those of anti-apoptotic Bcl-2. In mice exposed to radiation, geraniin treatment protected splenocytes and intestinal crypt cells from radiation-induced cell death. Our results suggest that geraniin presents radioprotective effects by regulating DNA damage on splenocytes, exerting immunostimulatory capacities and inhibiting apoptosis of radiosensitive immune cells and jejunal crypt cells. Therefore, geraniin can be a radioprotective agent against γ-irradiation exposure.     

Tocopherol succinate: A promising radiation countermeasure

Vitamin E is composed of a family of eight isomers known as tocols; consisting of four tocopherols and four tocotrienols that exist in four isomeric forms: alpha (α), beta (β), gamma (γ), and delta (δ). Earlier we have demonstrated the radioprotective efficacy of the succinate ester of α-tocopherol (α-tocopherol succinate, TS). CD2F1 mice were injected subcutaneously with 400 mg/kg of TS and irradiated with different doses of ⁶⁰Co γ-radiation to determine its radioprotective efficacy. The dose reduction factor (DRF) for TS was also determined. We also investigated effects of TS on cytokine production by multiplex Luminex and message by quantitative RT-PCR. Peripheral blood cells were enumerated from irradiated (3 and 7 Gy) and non-irradiated mice. TS significantly protected mice against lethal doses of ⁶⁰Co γ-radiation and the DRF was 1.28. TS stimulated granulocyte-colony stimulating factor (G-CSF) with a peak at 24 h after drug injection, and also stimulated G-CSF message as judged by RT-PCR in bone marrow cells at 12 and 24 h after injection. Further, pancytopenia studies revealed that TS significantly reduced thrombocytopenia, neutropenia, and monocytopenia. TS had no significant effect on lymphocytes indicating that it may be helpful only for the myeloid cell compartments. The stimulation of G-CSF by TS supports its effects on myeloid cell compartments. Our studies indicate that TS may be developed as a radioprotectant for humans against the potentially lethal effects of radiation exposure.     

A comparative evaluation of the influence of the complex drug Ukrain and its components on the effects of radiation

The development of cancer therapy is associated with an increased application of therapeutic schedules that combine radiation and different drugs. Ukrain is a promising drug proved to be efficient in the treatment of different tumors when used either before, during or after radiation therapy. Therefore, the issue of the influence of this drug on the effects of radiation is of interest. In previous studies, the ability of Ukrain to ameliorate the harmful consequences of irradiation was demonstrated. Ukrain is a semi-synthetic thiophosphoric derivative of Chelidonium majus L. alkaloids, i.e., it is a complex preparation. Therefore, it is necessary to elucidate which of its components are the active agents that mediate its antiradiation effects. The aim of this study was to compare the radioprotective effects of Ukrain and its components in experiments with animals irradiated by doses within a broad range (from LD50/30 to LD100/30). A total of 1,240 outbred male white mice were gamma-irradiated with an IGUR apparatus using 137Cs as the radiation source. Doses applied ranged from 5.25-7.5 Gy with a 0.75 increment between dosages. Dose rate was 1.41 Gy/min. The experiments have shown that the radioprotective effects of Ukrain are far superior to all its components taken separately, both measured by survival of mice irradiated by different doses, and by the protection coefficient a. These observations suggest that the influence of Ukrain on radiation effects does not result from the antiradiation properties of its components but rather from the concerted action of the specific combination found in Ukrain.     

A semiquinone glucoside derivative provides protection to male reproductive system of the mice against gamma radiation toxicity

Present investigation was carried out to evaluate the radioprotective efficacy of a novel Semiquinone glucoside derivative (SQGD), isolated from Bacillus sp. INM‐1, in the male reproductive system of BALB/c mice. Animals were administered 50 mg/kg b.wt. (i.p.) SQGD 2 h before whole body γ‐irradiation (10 Gy). Radiation‐induced cellular toxicity and its modulation by SQGD pretreatment was evaluated in the mice testes by quantitative histological and protein expression analysis. SQGD pretreatment protects irradiated mice from radiation‐induced testicular atrophy and germ cells degeneration, which may lead to emptiness of seminiferous tubules. Significant decrease in P53 and P21^{(Cip/WAF‐1)} expression was observed in the irradiated mice pretreated (2 h) by SQGD at 6 h compared with only irradiated mice. However, contrary to P53, expressions of P21 at latter time, that is, 24–72 h was found to be increased significantly in the irradiated mice pretreated by SQGD. Significant increase in the intact PARP‐1 protein expression were observed in the testes of the mice pretreated by SQGD 2 h before irradiation at 24–72 h compared with the only irradiated mice, whereas significant increase in PARP‐1 cleaved fragment was noticed at 24 h. Similarly, significant increase in NF‐kB and BCL‐2/BAX expressions ratio was noticed in SQGD‐treated mice (± irradiation) compared with irradiated mice, suggested a role of SQGD in the activation of prosurvival signaling in the testicular germinal cells population of the irradiated mice and thus contributed to protection against lethal γ‐irradiation. © 2012 Wiley Periodicals, Inc. Environ Toxicol 29: 558–567, 2014.