CN111184723B - Dsf-1在制备抗肿瘤和抗辐射药物中的应用 - Google Patents
Dsf-1在制备抗肿瘤和抗辐射药物中的应用 Download PDFInfo
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Abstract
本发明公开了DSF‑1在制备抗肿瘤和抗辐射药物中的应用,属于生物医药技术领域。本发明公开了DSF‑1在制备抗肿瘤和抗辐射药物中的应用以及双硫仑衍生物DSF‑1的制备方法。本发明公开的DSF‑1在制备抗肿瘤和抗辐射药物中的应用,结果证实DSF‑1有较好的抗肿瘤效果和辐射防护效果。
Description
技术领域
本发明涉及生物医药技术领域,更具体的说是涉及双硫仑衍生物DSF-1在制备抗肿瘤和抗辐射药物中的应用。
背景技术
双硫仑(disulfiram,DSF),分子式为C10H20N2S4,化学名为二硫化四乙基秋兰姆,又称戒酒硫、酒畏等,商品名为安塔布司(Antabuse),是一种较有前途的抗癌药物。早先发现DSF可以用于治疗酒精依赖,抑制酒精代谢过程中的乙醛脱氢酶,导致毒性物质乙醛在人体内大量累积,引起机体多种不适,达到戒酒目的;并且具有良好的安全性和耐受性。
有报道显示,DSF对多种肿瘤细胞株具有良好的体外抗肿瘤活性,如乳腺癌、黑色素瘤、结直肠癌、神经胶质瘤、肺癌和前列腺癌等。DSF抑制泛素蛋白酶体/核转录因子NFκB通路、多药耐药基因MDR1的表达、拓扑异构酶、基质金属蛋白酶、核蛋白定位因子NPL4,调控MAP激酶通路。DSF在体内的代谢物之一为二乙基二硫代氨基甲酸酯DDTC,包含自由巯基可以给出氢原子还原自由基,是一种很有潜力的抗氧化清除剂,直接应用DDTC具有辐射防护作用。国外研究证实双硫仑能够清除机体内自由基,能够保护r射线造成的质粒DNA和微粒体膜损伤,减轻大鼠和小鼠肝脏微粒体脂质过氧化反应;而且可以保护DNA免受芬顿效应所产生的羟自由基带来的放射损伤。然而DSF在体内代谢途径多,不稳定,DDTC的浓度很低,这导致了DSF在辐射防护作用上的研究受限。
辐射分为电离辐射和非电离辐射。其中,电离辐射容易对人体造成严重损伤。在电离辐射穿透物质的过程中,会产生许多复杂的物理及化学效应,包括:电离效用、热效用、荧光效用和光化学效用。与此同时,电离辐射又具有辐射生物效应,能够杀伤机体细胞,造成生物组织的破坏。辐射损伤先是使生物体内发生电离、激发及生成自由基,自由基的化学性质活泼,有极强的氧化应激损伤能力,可使各种生物膜的不饱和脂肪酸发生过氧化反应,形成脂质过氧化物,这些物质的形成具有一定的毒性,能使蛋白质分子内和分子间发生交联,比如去氢、加成、裂解等反应,从而导致DNA的双螺旋交联出现错误或者无法进行分裂,使得细胞周期发生紊乱,造成细胞变形、破坏和死亡。与此同时,过多的自由基会导致体内一些生物酶的活性发生改变,影响细胞的正常代谢,产生代谢功能紊乱,从而使生物体产生免疫、神经和内分泌系统的调节功能的障碍,可造成长远的损害。体内有自身一些抗氧化的蛋白和小分子,诸如,超氧化物歧化酶(Superoxide dismutase,SOD)、谷胱甘肽(Glutathione,GSH)、过氧化氢酶(Catalase,CAT)以及维生素A、C、E等,可以对自由基起到一定的清除作用。另一方面,电离辐射可直接作用于细胞核内的DNA生物大分子,造成DNA分子断裂或碱基的配对错误,引起急性辐射损伤。因体内自我调节的蛋白质和小分子对于辐射瞬间大量产生的自由基的清除和急性DNA损伤作用非常微弱,需要通过外源性抗氧化剂的协助,清除机体中过量的自由基,阻止细胞氧化应激,修复DNA损伤以维持机体的正常状态。
因此,对DSF结构进行改造,提供双硫仑衍生物DSF-1在制备抗肿瘤和抗辐射药物中的应用是本领域技术人员亟需解决的问题。
发明内容
有鉴于此,本发明提供了双硫仑衍生物DSF-1在制备抗肿瘤和抗辐射药物中的应用。
为了实现上述目的,本发明采用如下技术方案:
双硫仑衍生物DSF-1在制备抗肿瘤和抗辐射药物中的应用。
进一步,所述双硫仑衍生物DSF-1的制备方法如下:
取2mL甲基哌嗪,溶于乙醇;加入1mL浓度为0.72g/mL的NaOH溶液;冰水浴下缓慢滴加1.1mL二硫化碳,溶液起初为淡绿色,很快变为土黄色;室温搅拌6h,体系为棕黄色;将2mL含有2.3g碘单质的乙醇溶液分次加入,析出白色沉淀,得到双硫仑衍生物DSF-1。
进一步,所述双硫仑衍生物DSF-1的结构式如下:
经由上述的技术方案可知,与现有技术相比,本发明公开提供了DSF-1在制备抗肿瘤和抗辐射药物中的应用,DSF-1有较好的抗肿瘤效果和辐射防护效果。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据提供的附图获得其他的附图。
图1附图为本发明DSF-1的合成路线图;
图2附图为本发明DSF在有无FBS时对MCF7细胞的增殖抑制作用;
图3附图为本发明DSF在有无FBS时对A549细胞的增殖抑制作用;
图4附图为本发明DSF-1在有无FBS时对MCF7细胞的增殖抑制作用;
图5附图为本发明DSF-1在有无FBS时对A549细胞的增殖抑制作用;
图6附图为本发明DSF在有无NAC时对MCF7细胞的增殖抑制作用;
其中,*P<0.05,**P<0.01;
图7附图为本发明DSF在有无NAC时对A549细胞的增殖抑制作用;
其中,**P<0.01;
图8附图为本发明DSF-1在有无NAC时对MCF7细胞的增殖抑制作用;
其中,**P<0.01;
图9附图为本发明DSF-1在有无NAC时对A549细胞的增殖抑制作用;
其中,**P<0.01;
图10附图为本发明未添加DSF、DSF-1以及NAC溶液的MCF7细胞流式图;空白对照;
图11附图为本发明MCF7细胞加入30μmol/L的DSF溶液,作用24h后的细胞流式图;
图12附图为本发明MCF7细胞加入30μmol/L DSF和NAC的混合液,作用24h后的细胞流式图;
图13附图为本发明MCF7细胞加入10μmol/L DSF-1溶液,作用24h后的细胞流式图;
图14附图为本发明MCF7细胞加入10μmol/L DSF-1和NAC的混合液,作用24h后的细胞流式图;
图15附图为本发明未添加DSF、DSF-1以及NAC溶液的A549细胞流式图;空白对照;
图16附图为本发明A549细胞加入50μmol/L的DSF溶液,作用24h后的细胞流式图;
图17附图为本发明A549细胞加入50μmol/L DSF和NAC的混合液,作用24h后的细胞流式图;
图18附图为本发明A549细胞加入20μmol/L DSF-1溶液,作用24h后的细胞流式图;
图19附图为本发明A549细胞加入20μmol/L DSF-1和NAC的混合液,作用24h后的细胞流式图;
图20附图为本发明DSF-1作用于MCF-7细胞的凋亡百分比;
图21附图为本发明DSF-1作用于A549细胞的凋亡百分比;
图22附图为本发明Western Blot检测DSF-1作用于MCF7细胞后凋亡蛋白表达结果;
图23附图为本发明DSF-1作用于MCF7细胞后,PARP相对表达量结果;
图24附图为本发明DSF-1作用于MCF7细胞后,BAX相对表达量结果;
图25附图为本发明Western Blot检测DSF-1作用于A549细胞后凋亡蛋白表达结果;
图26附图为本发明DSF-1作用于A549细胞后,PARP相对表达量结果;
图27附图为本发明DSF-1作用于A549细胞后,BAX相对表达量结果;
图28附图为本发明致死剂量照射小鼠30天存活率结果;
图29附图为本发明小鼠外周血白细胞WBC数量;
图30附图为本发明小鼠外周血红细胞RBC数量;*P<0.05;
图31附图为本发明小鼠外周血血小板PLT数量;
图32附图为本发明小鼠外周血血红蛋白HGB含量;*P<0.05;
图33附图为本发明小鼠脾脏和胸腺指数;*P<0.05;
图34附图为本发明小鼠性腺指数;
图35附图为本发明小鼠肺和胰腺指数;
图36附图为本发明小鼠脾结节个数;
图37附图为本发明小鼠股骨有核细胞含量;
图38附图为本发明小鼠骨髓DNA含量;
图39附图为本发明小鼠的肝组织GSH含量;*P<0.05。
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
A549人肺癌细胞和MCF-7人乳腺癌细胞均由北京协和医学院基础研究所提供。DSF(纯度>99%)、N-乙酰-L-半胱氨酸(N-acetyl-L-cysteine,NAC)、Cell Counting Kit-8(CCK-8)试剂盒购自大连美仑生物公司;胎牛血清(Fetal Bovine Serum,FBS)购自以色列BI公司;磷脂酰丝氨酸蛋白抗体-碘化吡啶Annexin V-FITC/PI双染细胞凋亡检测试剂盒购自凯基生物有限公司;PARP抗体和BAX抗体购自美国proteintech公司;抗氧化试剂盒购自南京建成科技有限公司。
所有实验均三次重复,用统计软件Graphpad Prism 8进行数据分析及T检验,α=0.05为检验水准。
实施例1
双硫仑衍生物DSF-1的制备方法如下:
取2mL甲基哌嗪,溶于乙醇;加入1mL浓度为0.72g/mL的NaOH溶液;冰水浴下缓慢滴加1.1mL二硫化碳,溶液起初为淡绿色,很快变为土黄色;室温搅拌6h,体系为棕黄色;将2mL含有2.3g碘单质的乙醇溶液分次加入,很快有白色沉淀析出,反应4h体系为淡黄色,最后得到白色粉末2.085g,产率33.04%。合成路线见图1。
薄层层析(二氯甲烷:甲醇=8:1)显示为1个点;纯化后通过质谱和核磁氢谱、碳谱确证化合物的结构,为双(4-甲基-1-1-哌嗪基硫代甲酰基)二硫。
DSF-1为白色固体,熔点、氢谱、碳谱和质谱数据分别如下:
m.p.143.6~144.8℃;
1H NMR(CDCl3,
400MHz,)δ:2.38(s,6H,CH3),2.61[t,8H,N(CH2)2],4.34[t,8H,CSN(CH2)2];
13C NMR(CDCl3)δ:193.55(CS2),54.53(NCCN),45.61(CH3);
LC-MS m/z:351.081(M+).
抗肿瘤作用
实施例2细胞增殖情况检测
DSF和DSF-1分别用DMSO溶解后,用DMEM培养基将其稀释为50、10、5、1和0.5μmol/L的溶液;用含10%FBS的DMEM培养基将其稀释为10、5、1、0.5、0.1μmol/L的溶液。NAC用含10%FBS的DMEM培养基溶解,浓度为1mmol/L。
细胞处于对数生长期时胰酶消化,用细胞计数板计算细胞密度,按每孔5000个细胞接种于3个96孔板中,孵育24h。待细胞贴壁并稳定生长后弃去旧培养液,第一个96孔板分别加入含10%FBS的DMEM配置的不同浓度的DSF-1溶液100μL、含10%FBS的DMEM配置的不同浓度的DSF溶液100μL,空白孔加入10%FBS的DMEM。第二个96孔板先分别加入含10%FBS的DMEM配置的不同浓度的DSF-1溶液50μL、含10%FBS的DMEM配置的不同浓度的DSF溶液50μL,每个孔再加入配好的NAC溶液50μL混匀,空白孔为10%FBS的DMEM和NAC溶液的混合液。第三个96孔板分别加入DMEM配置的不同浓度的DSF-1溶液100μL、DMEM配置的不同浓度的DSF溶液100μL,空白组为DMEM。每个浓度均设3个复孔,继续培养48h后加入CCK-8溶液,37℃避光孵育,经酶标仪在480nm处检测吸光度值,利用Graphpad软件计算IC50值,绘制存活率图,并进行t检验。细胞存活率为试验孔吸光度值与对照孔吸光度值的比值。实验经三次重复。
10%FBS的DMEM培养基配置的DSF和DSF-1溶液作用于细胞(MCF7细胞、A549细胞)的结果见表1。作用于细胞48h时,DSF-1对MCF7细胞的IC50值在10μmol/L以下,优于DSF;对A549细胞的IC50值在30μmol/L以下,优于DSF。
表1 DSF和DSF-1分别作用于两种肿瘤细胞48h的IC50值
注:DSF-1与DSF通过配对t-检验分析比较有显著性差异,**P<0.01。
DSF在没有FBS存在时,对MCF7细胞的增殖抑制作用减弱,见图2。体系中没有FBS时,10μmol/L和50μmol/L的DSF作用的A549细胞的存活率与有FBS时相比有增加趋势,见图3。由于FBS中有铜离子,这一结果提示DSF抑制这两种肿瘤细胞的增殖需要铜离子的存在。与体系中有FBS相比,10μmol/L DSF-1作用的MCF7细胞在没有FBS时存活率显著增加,见图4。10μmol/L和50μmol/L DSF-1作用的A549细胞在没有FBS时存活率也可见增加,见图5。提示DSF-1发挥抗肿瘤作用需要FBS中铜离子的存在。
此外,NAC可以抑制ROS的产生,DSF通过产生ROS抑制肿瘤细胞的增殖。在MCF7细胞中1μmol/L的DSF中加入NAC与不加NAC相比存活率提高了约30%,见图6;在A549细胞中10μmol/L和50μmol/L的DSF中加入NAC与不加NAC相比存活率提高了约40%,见图7。10μmol/L的DSF-1对MCF7细胞的增殖抑制作用在加入NAC后明显减弱,差异具有统计学意义,见图8;在A549细胞中10μmol/L和50μmol/L的DSF-1中加入NAC与不加NAC相比存活率提高了约50%,见图9。提示DSF-1也是通过产生ROS发挥抑制肿瘤细胞增殖的作用。
实施例3细胞凋亡率检测
DSF和DSF-1分别用DMSO溶解后,用含10%FBS的DMEM培养基将DSF稀释为50、30μmol/L的溶液,将DSF-1稀释为20、10μmol/L的溶液。NAC用含10%FBS的DMEM培养基溶解,浓度为1mmol/L。对数生长期的A549细胞和MCF7细胞进行消化离心计数,六孔板每孔铺20万个细胞,孵育24h。弃去培养基,MCF7细胞各孔对应加入2mL 30μmol/L的DSF溶液,2mL 10μmol/L DSF-1溶液,1mL 30μmol/L DSF和1mL NAC的混合液,1mL 10μmol/L DSF-1和1mL NAC的混合液。A549细胞中各孔对应加入2mL 50μmol/L的DSF溶液,2mL 20μmol/L的DSF-1溶液,1mL50μmol/L DSF和1mL NAC的混合液,1mL 20μmol/L DSF-1和1mL NAC的混合液。
作用24h后收集死亡细胞,其余细胞用PBS冲洗并用胰酶消化收集并离心。将离心后的细胞重新悬浮在500μl缓冲液中,依次加入5μl FITC和5μl PI避光反应15min。然后上流式细胞仪检测细胞凋亡情况。MCF7细胞流式图结果见图10-图14;A549细胞流式图结果见图15-图19。用BD Accuri C6软件进行分析,直接可得细胞凋亡率。实验经三次重复。统计DSF-1作用于MCF-7细胞的凋亡百分比,结果见图20;图20结果表明,与30μmol/L的DSF相比,10μmol/L的DSF-1作用后的MCF7细胞的凋亡比例增加了约22%,有显著性差异;加入NAC后,10μmol/L的DSF-1作用后的MCF7细胞凋亡比例减少,有显著性差异。统计DSF-1作用于A549细胞的凋亡百分比,结果见图21;图21结果表明,与50μmol/L的DSF相比,20μmol/L的DSF-1作用后的A549细胞的凋亡比例增加了约9%,有显著性差异;加入NAC后,20μmol/L的DSF-1作用后的A549细胞凋亡比例减少,有显著性差异。
实施例4 Western Blot法检测
取对数生长期的A549细胞和MCF7的细胞消化离心计数,六孔板每孔铺20万个细胞,孵育24h。弃去培养基。DSF、DSF-1分别用DMSO溶解后,用含10%FBS的DMEM培养基稀释。MCF7细胞中各孔对应加入2mL 30μmol/L DSF溶液,2mL 10μmol/L DSF-1溶液,对照孔细胞(对照组)只加入10%FBS的DMEM培养基;A549细胞中各孔对应加入2mL 50μmol/L DSF溶液,2mL 20μmol/L DSF-1溶液,对照孔细胞(对照组)只加入10%FBS的DMEM培养基;作用24h后,收集细胞,常规裂解制备蛋白样品,经BCA蛋白定量法检测蛋白浓度。每组各取40μg总蛋白进行SDS-PAGE,随后将蛋白印迹转到PVDF膜上,经5%脱脂奶粉封闭,加入一抗溶液(1:1000稀释)4℃孵育过夜,PBST洗膜三次,PBS洗一次;加入相应二抗溶液(1:3000稀释),室温放置1h,洗三遍(前2遍用PBST洗,后1遍用PBS洗);最后采用ECL化学发光显色试剂孵育曝光,相关结果采用多色荧光化学发光成像分析系统进行拍照,Image J分析条带灰度值。蛋白相对表达量=目的蛋白灰度值/内参蛋白灰度值,实验经三次重复。
当一抗为PARP1兔多克隆抗体,对应二抗为辣根过氧化物酶标记山羊抗兔lgG(H+L);当一抗为BAX兔多克隆抗体,对应二抗为辣根过氧化物酶标记山羊抗兔lgG(H+L);当一抗为Beta Tublin鼠单克隆抗体,对应二抗为辣根过氧化物酶标记山羊抗鼠lgG(H+L)。
Western Blot检测DSF-1作用于MCF7细胞后凋亡蛋白表达结果见图22。DSF-1作用于MCF7细胞后,PARP相对表达量结果见图23;图23结果表明,与对照组相比,DSF-1作用后MCF7细胞的PARP表达水平降低(p<0.01)。DSF-1作用于MCF7细胞后,BAX相对表达量结果见图24;图24结果表明,与对照组相比,DSF及DSF-1作用于MCF7细胞后,BAX表达水平升高(p<0.01)。
Western Blot检测DSF-1作用于A549细胞后凋亡蛋白表达结果见图25。DSF-1作用于A549细胞后,PARP相对表达量结果见图26;图26结果表明,与对照组相比,DSF-1作用后A549细胞的PARP表达水平降低(p<0.05)。DSF-1作用于A549细胞后,BAX相对表达量结果见图27;图27结果表明,与对照组相比,DSF及DSF-1作用于A549细胞后,BAX表达水平升高(p<0.01)。
抗辐射作用
实施例5小鼠致死剂量30天存活率实验
DSF、DSF-1分组及灌胃给药
选用C57雄性小鼠40只,随机分4组,每组10只,设置空白对照组Control,单纯照射组Rad,DSF给药组,DSF-1给药组。化合物DSF、DSF-1均用羧甲基纤维素钠(CMC-Na)超声溶解,浓度为50mg/kg,灌胃给药。单纯照射组给予CMC-Na。照射前连续给药3d,照射当天照前30min给药,给予137Cs伽马射线7.2Gy一次性全身照射,剂量率为0.99Gy/min。每天观察各组小鼠的活动状态,记录30天内动物存活情况,graphpad做生存率的图。
致死剂量照射小鼠30天存活率实验结果见表2和图28。
表2照射小鼠30天存活率实验结果统计
注:与单纯照射组Rad相比较,*P<0.05,**P<0.01。
从表2和图28可以看出,与单纯照射组Rad相比,DSF-1提高辐射损伤小鼠的30d存活率约50%,延长辐射损伤小鼠的平均生存时间12天,差异具有统计学意义。提示DSF-1对C57具有较好的整体辐射防护效果。
实施例6小鼠亚致死剂量指标实验
1)试验动物分组及给药
DSF、DSF-1分组及灌胃给药
选用C57雄性小鼠18只,随机分3组,每组6只,设置单纯照射组,DSF给药组,DSF-1给药组。化合物DSF、DSF-1均用CMC-Na溶解,浓度为50mg/kg,灌胃给药。单纯照射组给予CMC-Na。照射前连续给药3d,照射当天照前30min给药,给予137Cs伽马射线6gy一次性全身照射,剂量率为0.99Gy/min。
2)检测指标
(1)血象分析
照射后第7d小鼠眼球取血200μL,加入到100μL的乙二胺四乙酸二钾盐(EDTA-2K)的抗凝溶液中,用全自动血细胞分析仪检测白细胞(White blood cells,WBC)、红细胞(Redblood cells,RBC)、血小板(Platelets,PLT)数和血红蛋白(hemoglobin,HGB)含量,并记录结果。
小鼠外周血白细胞WBC数量结果见图29;小鼠外周血红细胞RBC数量结果见图30;小鼠外周血血小板PLT数量结果见图31;小鼠外周血血红蛋白HGB含量结果见图32。与单纯照射组Rad相比,加入DSF-1对WBC、PLT均有一定程度提高,对RBC和HGB有微弱提高。提示DSF-1对受照小鼠的造血系统具有一定的保护作用。
(2)脏器指数
照射后第7d小鼠颈椎脱臼处死,解剖取小鼠脾脏、胸腺、性腺、肺、胰腺,用生理盐水冲洗干净血迹,精密电子天平称取质量,记录结果,按下列公式计算各脏器指数。
脏器指数=脏器质量(mg)/体重(g,解剖前的体重)。
小鼠脾脏和胸腺指数结果见图33;小鼠性腺指数结果见图34;小鼠肺和胰腺指数结果见图35。与单纯照射组Rad相比,DSF-1提高受照小鼠胸腺指数,具有统计学意义;也可在一定程度上提高肺,脾脏,胰腺及性腺指数。提示DSF-1对受照小鼠的脏器具有一定的保护作用。
(3)脾结节数(Colong forming unit of spleen,CFU-S)
照射后第7d将小鼠颈椎脱臼处死,解剖取脾脏放入Bouin氏液中固定,约24h后取出,用70%酒精冲洗去大部分黄色,肉眼计数脾脏表面突出的结节数(CFU-S)。小鼠脾结节个数结果见图36,表明DSF-1能够提高照射小鼠脾结节个数。提示DSF-1对受照小鼠的造血系统具有一定的保护作用。
(4)骨髓有核细胞数(Bone narrow nucleated cells,BMNC)
照射后第7d将小鼠颈椎脱臼处死,解剖小鼠,取左侧股骨,剔除全部肌肉组织,用生理盐水冲洗干净,剪去一端股骨头,另一端用7号针头刺穿股骨末端,用股骨有核细胞醋酸液反复冲洗股骨腔,直至股骨冲为白色为止(至少5遍)。用全自动血细胞分析仪检测股骨有核细胞数。小鼠股骨有核细胞含量结果见图37,表明DSF-1能够提高照射小鼠BMNC含量。提示DSF-1对受照小鼠的造血系统具有一定的保护作用。
(5)小鼠骨髓DNA含量
照射后第7d将小鼠颈椎脱臼处死,解剖小鼠,取另一侧股骨,剔除全部肌肉组织,用生理盐水冲洗干净,剪去一端的股骨头,另一端用2.5mL注射器针头刺穿股骨末端,用0.05mol/L氯化钙10mL反复冲洗骨髓腔,将全部骨髓冲入离心管中,直至股骨变为白色(至少冲洗5遍),冲出的骨髓液置于4℃冰箱放置30min,然后配平置于离心机中,2500r/min离心15min,弃上清液,向沉淀中加入0.2mol/L高氯酸5mL,充分混匀,90℃水浴加热15min,冷却后滤纸过滤,滤液用紫外分光光度计在268nm处测定吸光度OD值。小鼠骨髓DNA含量结果见图38,表明DSF-1能够提高照射小鼠骨髓DNA含量。提示DSF-1对受照小鼠的造血系统具有一定的保护作用。
实施例7受照射小鼠抗氧化试验
小鼠分组及给药情况与亚致死剂量指标实验(实施例6)分组一致。照射后第7天取肝脏组织,组织匀浆,用抗氧化试剂盒检测GSH指标。
与单纯照射组Rad相比,加药组(DSF、DSF-1)小鼠的肝组织GSH含量有一定增加,见图39。提示DSF-1对肝组织有一定的保护作用。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
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