本文主要研究内容
作者付瑞（2019）在《Th17/IL-17在小鼠血管内皮细胞衰老进程中的变化及意义》一文中研究指出：背景:衰老是自古以来人们一直在研究并去尝试减缓的问题,而在医学领域里,血管系统作为维持人体生命结构的重要组成之一,研究心血管系统的衰老具有着重要意义。血管内皮细胞分隔着血液成分,起着保护作用,同时其可分泌血管紧张素,以及表面的血小板依赖成分在机体的血管平衡稳态过程和抗凝过程发挥重要作用。血管内皮衰老后,形态学可见到细胞之间的空隙变大,细胞核和体积也增大且细胞形态不规则。衰老的血管结构不再完整,容易引起血管舒张及血栓形成,造成炎性物质的堆积,从而反馈调节又加重衰老。血管内皮的衰老对心血管疾病有着重要关系。辅助性T细胞17（helper T cells 17,Th17）是2007年新发现的一类不同于Th1,Th2的CD4+辅助性T细胞,可以分泌多种细胞因子,其中白细胞介素17（interleukin-17,IL-17）是Th17细胞的主要效应因子,Th17在先天免疫和获得性中起重要作用。并参与许多重要的生理或病理过程,如防御细胞外细菌感染,介导慢性炎症,器官移植的免疫排斥,自身免疫疾病和肿瘤的发作。目前的研究表明其在炎性反应、自身免疫性疾病等疾病中发挥重要作用。同时IL-17/Th17也可影响许多炎症因子的分泌与表达。而在小鼠体内,根据其CD44与CD62L的表达情况不同,可将其分为记忆性T细胞(CD4+CD44highCD62Llow)和初始T细胞(CD4+CD44lowCD62Lhigh)。衰老涉及到了包括CD4+T淋巴细胞在内的一些免疫细胞,而Th17细胞分泌白细胞介素-17,诱导炎症因子IL-6参与炎症反应,从而涉及到了炎症衰老这一假说,将Th17与衰老联系。衰老往往伴随着许多疾病（包括感染,癌症和一些自身免疫疾病）的发病率和严重程度的增加,导致生活质量下降,这些疾病都具有免疫成分的参与,免疫与衰老密切相关。血管内皮细胞的衰老和功能障碍参与动脉粥样硬化发生,内皮损伤被认为是动脉粥样硬化的第一个触发因素。此外,内皮细胞的功能障碍和细胞凋亡是斑块侵蚀和血栓形成的主要决定因素,导致急性冠脉综合征等疾病。。IL-17/Th17在血管内皮衰老进程中的作用及机制未有深入研究。在本课题中,检测Th17/IL-17在不同年龄段小鼠血清及脾脏中的表达情况,并通过培养小鼠主动脉原代内皮细胞,分析原代内皮细胞衰老通路变化和其他指标改变。同时检测小鼠血管动脉IL-17/IL-17RA表达情况,分析Th17/IL-17对血管内皮细胞衰老的影响,探讨其在血管内皮细胞衰老进程中的作用与意义,为心血管衰老提供新的方向。目的:研究小鼠原代血管内皮细胞衰老过程所涉及到的通路变化,分析在小鼠在不同年龄阶段其血管组织变化及脾脏和血清的Th17/IL-17表达变化,并分析其在血管内皮细胞衰老中产生的影响与意义。方法:（1）本研究首先以健康青年鼠、中年鼠、老年鼠为研究对象,每组20例,检测各年龄段小鼠血清IL-17浓度,以及相关细胞因子IL-6、细胞衰老相关分泌表型（senescent-associated secretory phenotype,SASP）因子IL-1β和动脉粥样硬化生化标志物Ox-LDL浓度均采用酶联免疫吸附法（Enzyme-linked immune-sorbent assay,ELISA）检测。并分析Th17/IL-17与这些SASP相关因子和炎症标志物表达水平的相关性。采用多参数流式细胞术检测技术,用不同的荧光色素抗体标记细胞,对小鼠脾脏所得的脾细胞悬液的淋巴细胞进行刺激和诱导实验得出Th17细胞的比率;检测CD44和CD62L在鼠Th17细胞中的表达情况;（2）不同年龄段小鼠收集主动脉做HE染色和免疫组化,观察衰老过程小鼠动脉血管变化以及IL17/17RA和内皮衰老指标VCAM-1在血管上的表达情况。（3）通过分离培养小鼠主动脉原代内皮细胞,经流式和免疫组化鉴定纯度,β-半乳糖苷酶染色检测原代细胞衰老情况,并检测衰老内皮细胞衰老相关异染色质位点（senescence-associated heterochromatic foci,SAHF）。并收集原代内皮细胞并提取蛋白,western-blot法检测内皮细胞衰老蛋白的表达变化,分析衰老所涉及的主要信号通路。结果:（1）老年鼠血清IL-17（210.82±27.68 pg/ml）及脾脏刺激诱导Th17细胞表达率（39.49±3.81%）显著高于健康青年鼠（88.78±14.35 pg/ml,23.53±4.74%）和健康中年鼠（128.47±16.63 pg/ml,27.28±3.86%）（均P<0.05）（2）青年鼠中年鼠老年鼠Th17细胞中记忆性T细胞(CD4+CD44highCD62Llow)细胞表达率分别为（82.57±6.578%）、（76.38±6.45%）、（78.54±6.87%）,（P>0.05）。Th17细胞初始T细胞表达率(CD4+CD44lowCD62LhighIL-17+/CD4+IL17+T细胞)在青年鼠（5.82±2.61%）,中年鼠（7.73±3.26%）,老年鼠（6.52±2.96%）,各组间表达差异无统计学意义（均P>0.05）。各组Th17细胞以记忆性T细胞为主。（3）老年鼠体内血清IL-6、IL-1β及动脉粥样硬化相关指标ox-LDL浓度明显高于青年鼠和中年鼠,且相关性分析表明,IL-17/Th17与这些指标变化呈有正相关性。（4）小鼠主动脉HE染色和免疫组化结果:主动脉HE染色结果显示青年鼠和中年鼠内皮细胞排列整齐,细胞之间连接紧密,内皮下间隙小。而老年鼠动脉较青年鼠和中年鼠血管壁增厚,内膜变粗糙,内皮间隙增宽,排列疏松。主动脉免疫组化结果显示,老年鼠动脉IL-17、IL-17RA和内皮衰老指标VCAM-1表达水平均明显高于青年鼠和中年鼠。（5）分离培养鼠原代内皮细胞,衰老染色结果显示,老年鼠原代血管内皮细胞具有阳性细胞表达率和衰老相关异位染色质（SAHF）表达明显增多,Western-Blot结果表明原代内皮细胞P16、P19、P21、P53和Rb等衰老蛋白蛋白表达明显增强。结论:（1）老年鼠Th17表达率、体内血清IL-17、IL-6浓度以及以及SASP因子IL-1β浓度较青年和中年鼠明显增高;Th17/IL-17的表达水平与IL-6、IL-1β浓度水平呈显著正相关。（2）老年鼠原代血管内皮衰老染色阳性率、SAHF表达率较青年和中年鼠明显增高。（3）衰老小鼠其主动脉IL-17及受体IL-17RA、血管内皮衰老指标VCAM-1表达明显增多。由此可见,Th17/IL-17在衰老进程中表达明显增强,与小鼠血管内皮细胞衰老密切相关,为衰老和心脑血管疾病研究提供新的研究点。
Abstract
bei jing :cui lao shi zi gu yi lai ren men yi zhi zai yan jiu bing qu chang shi jian huan de wen ti ,er zai yi xue ling yu li ,xie guan ji tong zuo wei wei chi ren ti sheng ming jie gou de chong yao zu cheng zhi yi ,yan jiu xin xie guan ji tong de cui lao ju you zhao chong yao yi yi 。xie guan nei pi xi bao fen ge zhao xie ye cheng fen ,qi zhao bao hu zuo yong ,tong shi ji ke fen bi xie guan jin zhang su ,yi ji biao mian de xie xiao ban yi lai cheng fen zai ji ti de xie guan ping heng wen tai guo cheng he kang ning guo cheng fa hui chong yao zuo yong 。xie guan nei pi cui lao hou ,xing tai xue ke jian dao xi bao zhi jian de kong xi bian da ,xi bao he he ti ji ye zeng da ju xi bao xing tai bu gui ze 。cui lao de xie guan jie gou bu zai wan zheng ,rong yi yin qi xie guan shu zhang ji xie shuan xing cheng ,zao cheng yan xing wu zhi de dui ji ,cong er fan kui diao jie you jia chong cui lao 。xie guan nei pi de cui lao dui xin xie guan ji bing you zhao chong yao guan ji 。fu zhu xing Txi bao 17（helper T cells 17,Th17）shi 2007nian xin fa xian de yi lei bu tong yu Th1,Th2de CD4+fu zhu xing Txi bao ,ke yi fen bi duo chong xi bao yin zi ,ji zhong bai xi bao jie su 17（interleukin-17,IL-17）shi Th17xi bao de zhu yao xiao ying yin zi ,Th17zai xian tian mian yi he huo de xing zhong qi chong yao zuo yong 。bing can yu hu duo chong yao de sheng li huo bing li guo cheng ,ru fang yu xi bao wai xi jun gan ran ,jie dao man xing yan zheng ,qi guan yi zhi de mian yi pai chi ,zi shen mian yi ji bing he zhong liu de fa zuo 。mu qian de yan jiu biao ming ji zai yan xing fan ying 、zi shen mian yi xing ji bing deng ji bing zhong fa hui chong yao zuo yong 。tong shi IL-17/Th17ye ke ying xiang hu duo yan zheng yin zi de fen bi yu biao da 。er zai xiao shu ti nei ,gen ju ji CD44yu CD62Lde biao da qing kuang bu tong ,ke jiang ji fen wei ji yi xing Txi bao (CD4+CD44highCD62Llow)he chu shi Txi bao (CD4+CD44lowCD62Lhigh)。cui lao she ji dao le bao gua CD4+Tlin ba xi bao zai nei de yi xie mian yi xi bao ,er Th17xi bao fen bi bai xi bao jie su -17,you dao yan zheng yin zi IL-6can yu yan zheng fan ying ,cong er she ji dao le yan zheng cui lao zhe yi jia shui ,jiang Th17yu cui lao lian ji 。cui lao wang wang ban sui zhao hu duo ji bing （bao gua gan ran ,ai zheng he yi xie zi shen mian yi ji bing ）de fa bing lv he yan chong cheng du de zeng jia ,dao zhi sheng huo zhi liang xia jiang ,zhe xie ji bing dou ju you mian yi cheng fen de can yu ,mian yi yu cui lao mi qie xiang guan 。xie guan nei pi xi bao de cui lao he gong neng zhang ai can yu dong mai yu yang ying hua fa sheng ,nei pi sun shang bei ren wei shi dong mai yu yang ying hua de di yi ge chu fa yin su 。ci wai ,nei pi xi bao de gong neng zhang ai he xi bao diao wang shi ban kuai qin shi he xie shuan xing cheng de zhu yao jue ding yin su ,dao zhi ji xing guan mai zeng ge zheng deng ji bing 。。IL-17/Th17zai xie guan nei pi cui lao jin cheng zhong de zuo yong ji ji zhi wei you shen ru yan jiu 。zai ben ke ti zhong ,jian ce Th17/IL-17zai bu tong nian ling duan xiao shu xie qing ji pi zang zhong de biao da qing kuang ,bing tong guo pei yang xiao shu zhu dong mai yuan dai nei pi xi bao ,fen xi yuan dai nei pi xi bao cui lao tong lu bian hua he ji ta zhi biao gai bian 。tong shi jian ce xiao shu xie guan dong mai IL-17/IL-17RAbiao da qing kuang ,fen xi Th17/IL-17dui xie guan nei pi xi bao cui lao de ying xiang ,tan tao ji zai xie guan nei pi xi bao cui lao jin cheng zhong de zuo yong yu yi yi ,wei xin xie guan cui lao di gong xin de fang xiang 。mu de :yan jiu xiao shu yuan dai xie guan nei pi xi bao cui lao guo cheng suo she ji dao de tong lu bian hua ,fen xi zai xiao shu zai bu tong nian ling jie duan ji xie guan zu zhi bian hua ji pi zang he xie qing de Th17/IL-17biao da bian hua ,bing fen xi ji zai xie guan nei pi xi bao cui lao zhong chan sheng de ying xiang yu yi yi 。fang fa :（1）ben yan jiu shou xian yi jian kang qing nian shu 、zhong nian shu 、lao nian shu wei yan jiu dui xiang ,mei zu 20li ,jian ce ge nian ling duan xiao shu xie qing IL-17nong du ,yi ji xiang guan xi bao yin zi IL-6、xi bao cui lao xiang guan fen bi biao xing （senescent-associated secretory phenotype,SASP）yin zi IL-1βhe dong mai yu yang ying hua sheng hua biao zhi wu Ox-LDLnong du jun cai yong mei lian mian yi xi fu fa （Enzyme-linked immune-sorbent assay,ELISA）jian ce 。bing fen xi Th17/IL-17yu zhe xie SASPxiang guan yin zi he yan zheng biao zhi wu biao da shui ping de xiang guan xing 。cai yong duo can shu liu shi xi bao shu jian ce ji shu ,yong bu tong de ying guang se su kang ti biao ji xi bao ,dui xiao shu pi zang suo de de pi xi bao xuan ye de lin ba xi bao jin hang ci ji he you dao shi yan de chu Th17xi bao de bi lv ;jian ce CD44he CD62Lzai shu Th17xi bao zhong de biao da qing kuang ;（2）bu tong nian ling duan xiao shu shou ji zhu dong mai zuo HEran se he mian yi zu hua ,guan cha cui lao guo cheng xiao shu dong mai xie guan bian hua yi ji IL17/17RAhe nei pi cui lao zhi biao VCAM-1zai xie guan shang de biao da qing kuang 。（3）tong guo fen li pei yang xiao shu zhu dong mai yuan dai nei pi xi bao ,jing liu shi he mian yi zu hua jian ding chun du ,β-ban ru tang gan mei ran se jian ce yuan dai xi bao cui lao qing kuang ,bing jian ce cui lao nei pi xi bao cui lao xiang guan yi ran se zhi wei dian （senescence-associated heterochromatic foci,SAHF）。bing shou ji yuan dai nei pi xi bao bing di qu dan bai ,western-blotfa jian ce nei pi xi bao cui lao dan bai de biao da bian hua ,fen xi cui lao suo she ji de zhu yao xin hao tong lu 。jie guo :（1）lao nian shu xie qing IL-17（210.82±27.68 pg/ml）ji pi zang ci ji you dao Th17xi bao biao da lv （39.49±3.81%）xian zhe gao yu jian kang qing nian shu （88.78±14.35 pg/ml,23.53±4.74%）he jian kang zhong nian shu （128.47±16.63 pg/ml,27.28±3.86%）（jun P<0.05）（2）qing nian shu zhong nian shu lao nian shu Th17xi bao zhong ji yi xing Txi bao (CD4+CD44highCD62Llow)xi bao biao da lv fen bie wei （82.57±6.578%）、（76.38±6.45%）、（78.54±6.87%）,（P>0.05）。Th17xi bao chu shi Txi bao biao da lv (CD4+CD44lowCD62LhighIL-17+/CD4+IL17+Txi bao )zai qing nian shu （5.82±2.61%）,zhong nian shu （7.73±3.26%）,lao nian shu （6.52±2.96%）,ge zu jian biao da cha yi mo tong ji xue yi yi （jun P>0.05）。ge zu Th17xi bao yi ji yi xing Txi bao wei zhu 。（3）lao nian shu ti nei xie qing IL-6、IL-1βji dong mai yu yang ying hua xiang guan zhi biao ox-LDLnong du ming xian gao yu qing nian shu he zhong nian shu ,ju xiang guan xing fen xi biao ming ,IL-17/Th17yu zhe xie zhi biao bian hua cheng you zheng xiang guan xing 。（4）xiao shu zhu dong mai HEran se he mian yi zu hua jie guo :zhu dong mai HEran se jie guo xian shi qing nian shu he zhong nian shu nei pi xi bao pai lie zheng ji ,xi bao zhi jian lian jie jin mi ,nei pi xia jian xi xiao 。er lao nian shu dong mai jiao qing nian shu he zhong nian shu xie guan bi zeng hou ,nei mo bian cu cao ,nei pi jian xi zeng kuan ,pai lie shu song 。zhu dong mai mian yi zu hua jie guo xian shi ,lao nian shu dong mai IL-17、IL-17RAhe nei pi cui lao zhi biao VCAM-1biao da shui ping jun ming xian gao yu qing nian shu he zhong nian shu 。（5）fen li pei yang shu yuan dai nei pi xi bao ,cui lao ran se jie guo xian shi ,lao nian shu yuan dai xie guan nei pi xi bao ju you yang xing xi bao biao da lv he cui lao xiang guan yi wei ran se zhi （SAHF）biao da ming xian zeng duo ,Western-Blotjie guo biao ming yuan dai nei pi xi bao P16、P19、P21、P53he Rbdeng cui lao dan bai dan bai biao da ming xian zeng jiang 。jie lun :（1）lao nian shu Th17biao da lv 、ti nei xie qing IL-17、IL-6nong du yi ji yi ji SASPyin zi IL-1βnong du jiao qing nian he zhong nian shu ming xian zeng gao ;Th17/IL-17de biao da shui ping yu IL-6、IL-1βnong du shui ping cheng xian zhe zheng xiang guan 。（2）lao nian shu yuan dai xie guan nei pi cui lao ran se yang xing lv 、SAHFbiao da lv jiao qing nian he zhong nian shu ming xian zeng gao 。（3）cui lao xiao shu ji zhu dong mai IL-17ji shou ti IL-17RA、xie guan nei pi cui lao zhi biao VCAM-1biao da ming xian zeng duo 。you ci ke jian ,Th17/IL-17zai cui lao jin cheng zhong biao da ming xian zeng jiang ,yu xiao shu xie guan nei pi xi bao cui lao mi qie xiang guan ,wei cui lao he xin nao xie guan ji bing yan jiu di gong xin de yan jiu dian 。
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论文作者分别是来自安徽医科大学的付瑞，发表于刊物安徽医科大学2019-09-02论文，是一篇关于血管内皮细胞论文,白细胞介素论文,细胞论文,细胞衰老论文,安徽医科大学2019-09-02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，资料来自安徽医科大学2019-09-02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。
标签：血管内皮细胞论文; 白细胞介素论文; 细胞论文; 细胞衰老论文; 安徽医科大学2019-09-02论文;