本文主要研究内容
作者付运星（2019）在《DPTM药效学和药代动力学研究》一文中研究指出：14-O-[（4,6-二氨基嘧啶-2-基）硫代乙酰基)姆体林（DPTM）为本课题组自主合成的一种新型截短侧耳素类衍生物。前期研究证明,DPTM在体外有较好的抑菌作用,然而其临床应用潜力尚且不知,因此本文进一步研究了该化合物的临床前药效学及药代动力学,其主要研究内容及成果如下:1.DPTM对小鼠多杀性巴氏杆菌（Pasteurella multocida）肺炎模型治疗作用研究首先在体外测定了DPTM对Pasteurella multocida的最小抑菌浓度（Minimum inhibitory concentration,MIC）及杀菌曲线。其次,将小鼠随机分为5组,每组15只,将每组小鼠腹腔注射进行麻醉,取60μL 4.0×109 CFU/mL的菌悬液缓慢对小鼠进行鼻腔内灌注,建立小鼠Pasteurella multocida肺炎感染模型,在肺炎感染模型的基础上,通过小鼠存活率和肺湿干重（W/D）比测定,肺组织病理学变化分析,肺部炎性细胞因子、以及肺和血液中Pasteurella multocida的生长变化检测,来评估DPTM对Pasteurella multocida的治疗作用。结果表明:DPTM在体外对Pasteurella multocida具有较好的抑制作用,其MIC为0.781μg/mL,DPTM在体内能显著性地降低小鼠的死亡率,减轻肺部病理损伤,降低肺部TNF-α,IL-6,IL-1β和MCP-1炎性细胞因子的分泌,减少细菌在肺部和血液中的载量。2.DPTM对耐甲氧西林金黄色葡萄球菌（Methicillin-resistant Staphylococcus aureus,MRSA）小鼠皮肤感染治疗作用研究小鼠随机分为6组,每组10只,用无菌剪刀在后背部正中脊柱约0.5 cm处作长约1.5 cm深度至肌膜作一竖型切口,取对数生长期的MRSA43300菌液0.1 mL(1012 CFU/mL)均匀涂抹于整个创面建立小鼠深层皮肤感模型。在建立MRSA43300小鼠皮肤局部感染模型的基础上,检测白细胞数量和细菌数量变化、血浆中炎性细胞因子IL-6,TNF-α,IL-1β和VEGF水平的变化、利用HE及Masson三色染色来评估DPTM对MRSA43300的治疗作用。结果表明:DPTM可促进MRSA43300皮肤感染的愈合,减少感染皮肤MRSA43300数量,降低血浆中的IL-6,TNF-α,IL-1β炎性细胞因子的分泌。3.亚抑菌浓度DPTM对MRSAα-溶血素的抑制作用研究通过测定MRSA43300与亚抑菌浓度的DPTM共培养后细菌上清液的溶血能力、检测α-溶血素的分泌水平和细菌的HIa和agrA mRNA转录水平,来研究DPTM在亚抑菌浓度时对MRSA43300α-溶血素的影响。结果表明:DPTM亚抑菌浓度可以抑制MRSA43300的α-溶血素的分泌。4.亚抑菌浓度DPTM对MRSA诱导RAW264.7巨噬细胞损伤的保护作用研究将亚抑菌浓度的DPTM、MRSA43300及RAW264.7巨噬细胞共培养,通过检测LDH和NO的释放、IL-6,IL-1β和TNF-α的分泌、Phospho-p65、iNOS和Cox-2蛋白表达量及NF-κB核转运,来评估DPTM对MRSA43300诱导细胞损伤的保护作用。结果表明:DPTM亚抑菌浓度能减轻MRSA43300诱导的RAW264.7细胞损伤。5.DPTM在大鼠体内的药代动力学、组织分布及尿液和粪便排泄研究DPTM（10、25和75mg/kg）以静脉注射,肌肉注射和口服三种方式给药后,运用HPLC-MS/MS技术,研究DPTM在大鼠体内的药代动力学,组织分布及尿液和粪便排泄特点。结果表明:DPTM可被大鼠快速吸收并快速清除,并广泛分布于所检测的各种组织中,主要通过粪便排出机体。6.DPTM对肝微粒体CYP450酶和基因表达的影响DPTM与大鼠肝微粒体CYP450酶或HepG2细胞共培养,通过Cocktail探针药物法及Real-timePCR研究DPTM对大鼠肝微粒体CYP450酶及HepG2细胞的CYP450s mRNA表达影响。结果表明:DPTM对CYP3A具有强烈的抑制作用;对CYP2E1酶活性有一定的抑制作用;而对CYP2D6、CYP1A2、CYP2C酶活性抑制不显著。此外,DPTM对HepG2细胞的CYP3A4、CYP2C9及CYP1A2的mRNA表达呈显著诱导作用,但随着孵育时间的延长,DPTM可显著抑制CYP2E1与CYP2D6的mRNA表达。总之,DPTM对Pasteurella multocida肺部感染小鼠具有保护作用,能明显减轻肺部病理损伤;DPTM软膏可促进MRSA43300皮肤感染的愈合,显著抑制MRSA43300α-溶血素的分泌且对MRSA43300诱导的RAW264.7巨噬细胞损伤具有保护作用;该药物具有良好的药代动力学参数,组织分布较为广泛,对CYP450酶具有选择性抑制或诱导作用,具有较大的临床开发应用价值。
Abstract
14-O-[（4,6-er an ji mi ding -2-ji ）liu dai yi xian ji )mu ti lin （DPTM）wei ben ke ti zu zi zhu ge cheng de yi chong xin xing jie duan ce er su lei yan sheng wu 。qian ji yan jiu zheng ming ,DPTMzai ti wai you jiao hao de yi jun zuo yong ,ran er ji lin chuang ying yong qian li shang ju bu zhi ,yin ci ben wen jin yi bu yan jiu le gai hua ge wu de lin chuang qian yao xiao xue ji yao dai dong li xue ,ji zhu yao yan jiu nei rong ji cheng guo ru xia :1.DPTMdui xiao shu duo sha xing ba shi gan jun （Pasteurella multocida）fei yan mo xing zhi liao zuo yong yan jiu shou xian zai ti wai ce ding le DPTMdui Pasteurella multocidade zui xiao yi jun nong du （Minimum inhibitory concentration,MIC）ji sha jun qu xian 。ji ci ,jiang xiao shu sui ji fen wei 5zu ,mei zu 15zhi ,jiang mei zu xiao shu fu qiang zhu she jin hang ma zui ,qu 60μL 4.0×109 CFU/mLde jun xuan ye huan man dui xiao shu jin hang bi qiang nei guan zhu ,jian li xiao shu Pasteurella multocidafei yan gan ran mo xing ,zai fei yan gan ran mo xing de ji chu shang ,tong guo xiao shu cun huo lv he fei shi gan chong （W/D）bi ce ding ,fei zu zhi bing li xue bian hua fen xi ,fei bu yan xing xi bao yin zi 、yi ji fei he xie ye zhong Pasteurella multocidade sheng chang bian hua jian ce ,lai ping gu DPTMdui Pasteurella multocidade zhi liao zuo yong 。jie guo biao ming :DPTMzai ti wai dui Pasteurella multocidaju you jiao hao de yi zhi zuo yong ,ji MICwei 0.781μg/mL,DPTMzai ti nei neng xian zhe xing de jiang di xiao shu de si wang lv ,jian qing fei bu bing li sun shang ,jiang di fei bu TNF-α,IL-6,IL-1βhe MCP-1yan xing xi bao yin zi de fen bi ,jian shao xi jun zai fei bu he xie ye zhong de zai liang 。2.DPTMdui nai jia yang xi lin jin huang se pu tao qiu jun （Methicillin-resistant Staphylococcus aureus,MRSA）xiao shu pi fu gan ran zhi liao zuo yong yan jiu xiao shu sui ji fen wei 6zu ,mei zu 10zhi ,yong mo jun jian dao zai hou bei bu zheng zhong ji zhu yao 0.5 cmchu zuo chang yao 1.5 cmshen du zhi ji mo zuo yi shu xing qie kou ,qu dui shu sheng chang ji de MRSA43300jun ye 0.1 mL(1012 CFU/mL)jun yun tu ma yu zheng ge chuang mian jian li xiao shu shen ceng pi fu gan mo xing 。zai jian li MRSA43300xiao shu pi fu ju bu gan ran mo xing de ji chu shang ,jian ce bai xi bao shu liang he xi jun shu liang bian hua 、xie jiang zhong yan xing xi bao yin zi IL-6,TNF-α,IL-1βhe VEGFshui ping de bian hua 、li yong HEji Massonsan se ran se lai ping gu DPTMdui MRSA43300de zhi liao zuo yong 。jie guo biao ming :DPTMke cu jin MRSA43300pi fu gan ran de yu ge ,jian shao gan ran pi fu MRSA43300shu liang ,jiang di xie jiang zhong de IL-6,TNF-α,IL-1βyan xing xi bao yin zi de fen bi 。3.ya yi jun nong du DPTMdui MRSAα-rong xie su de yi zhi zuo yong yan jiu tong guo ce ding MRSA43300yu ya yi jun nong du de DPTMgong pei yang hou xi jun shang qing ye de rong xie neng li 、jian ce α-rong xie su de fen bi shui ping he xi jun de HIahe agrA mRNAzhuai lu shui ping ,lai yan jiu DPTMzai ya yi jun nong du shi dui MRSA43300α-rong xie su de ying xiang 。jie guo biao ming :DPTMya yi jun nong du ke yi yi zhi MRSA43300de α-rong xie su de fen bi 。4.ya yi jun nong du DPTMdui MRSAyou dao RAW264.7ju shi xi bao sun shang de bao hu zuo yong yan jiu jiang ya yi jun nong du de DPTM、MRSA43300ji RAW264.7ju shi xi bao gong pei yang ,tong guo jian ce LDHhe NOde shi fang 、IL-6,IL-1βhe TNF-αde fen bi 、Phospho-p65、iNOShe Cox-2dan bai biao da liang ji NF-κBhe zhuai yun ,lai ping gu DPTMdui MRSA43300you dao xi bao sun shang de bao hu zuo yong 。jie guo biao ming :DPTMya yi jun nong du neng jian qing MRSA43300you dao de RAW264.7xi bao sun shang 。5.DPTMzai da shu ti nei de yao dai dong li xue 、zu zhi fen bu ji niao ye he fen bian pai xie yan jiu DPTM（10、25he 75mg/kg）yi jing mai zhu she ,ji rou zhu she he kou fu san chong fang shi gei yao hou ,yun yong HPLC-MS/MSji shu ,yan jiu DPTMzai da shu ti nei de yao dai dong li xue ,zu zhi fen bu ji niao ye he fen bian pai xie te dian 。jie guo biao ming :DPTMke bei da shu kuai su xi shou bing kuai su qing chu ,bing an fan fen bu yu suo jian ce de ge chong zu zhi zhong ,zhu yao tong guo fen bian pai chu ji ti 。6.DPTMdui gan wei li ti CYP450mei he ji yin biao da de ying xiang DPTMyu da shu gan wei li ti CYP450mei huo HepG2xi bao gong pei yang ,tong guo Cocktailtan zhen yao wu fa ji Real-timePCRyan jiu DPTMdui da shu gan wei li ti CYP450mei ji HepG2xi bao de CYP450s mRNAbiao da ying xiang 。jie guo biao ming :DPTMdui CYP3Aju you jiang lie de yi zhi zuo yong ;dui CYP2E1mei huo xing you yi ding de yi zhi zuo yong ;er dui CYP2D6、CYP1A2、CYP2Cmei huo xing yi zhi bu xian zhe 。ci wai ,DPTMdui HepG2xi bao de CYP3A4、CYP2C9ji CYP1A2de mRNAbiao da cheng xian zhe you dao zuo yong ,dan sui zhao fu yo shi jian de yan chang ,DPTMke xian zhe yi zhi CYP2E1yu CYP2D6de mRNAbiao da 。zong zhi ,DPTMdui Pasteurella multocidafei bu gan ran xiao shu ju you bao hu zuo yong ,neng ming xian jian qing fei bu bing li sun shang ;DPTMruan gao ke cu jin MRSA43300pi fu gan ran de yu ge ,xian zhe yi zhi MRSA43300α-rong xie su de fen bi ju dui MRSA43300you dao de RAW264.7ju shi xi bao sun shang ju you bao hu zuo yong ;gai yao wu ju you liang hao de yao dai dong li xue can shu ,zu zhi fen bu jiao wei an fan ,dui CYP450mei ju you shua ze xing yi zhi huo you dao zuo yong ,ju you jiao da de lin chuang kai fa ying yong jia zhi 。
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论文作者分别是来自中国农业科学院的付运星，发表于刊物中国农业科学院2019-07-05论文，是一篇关于药代动力学论文,中国农业科学院2019-07-05论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自中国农业科学院2019-07-05论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。