J9九游会AG

J9九游会AG

恭喜!国家纳米中心使用CULTEX细胞暴露技术研究空气污染的高分文章发表

恭喜!国家纳米中心使用CULTEX细胞暴露技术研究空气污染的高分文章发表

2019年(nian)2月,来自于中(zhong)国科(ke)(ke)学(xue)院(yuan)国家纳米中(zhong)心、山西大学(xue)环(huan)境与(yu)健康(kang)研究中(zhong)心环(huan)境与(yu)资源学(xue)院(yuan)、中(zhong)国科(ke)(ke)学(xue)院(yuan)大学(xue)的(de)多位科(ke)(ke)学(xue)家在杰(jie)出的(de)环(huan)境类期刊Chemosphere发表题为“Complex to simple: In vitro exposure of particulate matter simulated at the air-liquid interface discloses the health impacts of major air pollutants”关于复合空气污染物体外气液界(jie)面毒性(xing)研究的(de)进展。

空气颗粒物(PM):

PM的(de)(de)(de)主要无机成(cheng)(cheng)分(fen)包括硫(liu)(liu)酸盐、硝(xiao)酸盐、铵、氯化(hua)物(wu)和微量金属(Wang等(deng)(deng)(deng)人(ren),2013年(nian);Han等(deng)(deng)(deng)人(ren),2016年(nian)),其(qi)(qi)中硫(liu)(liu)酸盐对人(ren)体健康的(de)(de)(de)危害(hai)比(bi)其(qi)(qi)他PM成(cheng)(cheng)分(fen)更大(da)(Ueda等(deng)(deng)(deng)人(ren),2016年(nian)),这些硫(liu)(liu)酸盐主要是(shi)由人(ren)类排放到大(da)气(qi)(qi)(qi)(qi)(qi)中的(de)(de)(de)SO2 在一定条件下经(jing)复(fu)杂(za)的(de)(de)(de)氧(yang)化(hua)还原反应(ying)(ying)形(xing)(xing)成(cheng)(cheng),同(tong)时(shi)吸附到气(qi)(qi)(qi)(qi)(qi)溶胶(jiao)颗(ke)粒(li)表(biao)面(mian)与其(qi)(qi)他物(wu)质一起形(xing)(xing)成(cheng)(cheng)复(fu)杂(za)的(de)(de)(de)协同(tong)毒性(xing)作(zuo)用。二氧(yang)化(hua)硅(SiO2)是(shi)大(da)气(qi)(qi)(qi)(qi)(qi)气(qi)(qi)(qi)(qi)(qi)溶胶(jiao)的(de)(de)(de)主要成(cheng)(cheng)分(fen)之(zhi)一,由于(yu)其(qi)(qi)结构稳定、比(bi)表(biao)面(mian)积大(da)、毒性(xing)相对较(jiao)低,被用来(lai)模拟(ni)大(da)气(qi)(qi)(qi)(qi)(qi)粒(li)子的(de)(de)(de)核心(Wang等(deng)(deng)(deng)人(ren),2018)。同(tong)时(shi),随着纳米技术的(de)(de)(de)发展(zhan),二氧(yang)化(hua)硅纳米颗(ke)粒(li)的(de)(de)(de)应(ying)(ying)用也越来(lai)越广(guang)泛。之(zhi)前(qian)的(de)(de)(de)研(yan)究表(biao)明(ming),空(kong)气(qi)(qi)(qi)(qi)(qi)中的(de)(de)(de)二氧(yang)化(hua)硫(liu)(liu)可以被吸附到大(da)气(qi)(qi)(qi)(qi)(qi)颗(ke)粒(li)物(wu)的(de)(de)(de)表(biao)面(mian)(Han等(deng)(deng)(deng)人(ren),2016年(nian))。因此,通过模拟(ni)硫(liu)(liu)酸盐吸附在SiO2上形(xing)(xing)成(cheng)(cheng)的(de)(de)(de)气(qi)(qi)(qi)(qi)(qi)溶胶(jiao),可用于(yu)研(yan)究空(kong)气(qi)(qi)(qi)(qi)(qi)污染物(wu)的(de)(de)(de)毒性(xing)效应(ying)(ying)和健康风险(xian)。

CULTEX气-液界(jie)面暴露技术:

到目前为(wei)止,还没(mei)(mei)有(you)(you)使(shi)用(yong)(yong)(yong)体(ti)外模(mo)(mo)(mo)型来研(yan)(yan)究颗(ke)粒物(wu)(wu)与二氧(yang)化硫的(de)(de)(de)协同毒性(xing)(xing)(xing)(xing)作(zuo)用(yong)(yong)(yong),主要(yao)是(shi)由(you)(you)于(yu)(yu)缺乏合适颗(ke)粒物(wu)(wu)细胞体(ti)外暴露(lu)的(de)(de)(de)系(xi)统。传统浸没(mei)(mei)式暴露(lu)实(shi)验设(she)计中(zhong)(zhong),直(zhi)接(jie)将(jiang)颗(ke)粒添(tian)加到细胞培养基(ji)中(zhong)(zhong)评估颗(ke)粒毒性(xing)(xing)(xing)(xing)。然而,这些方法有(you)(you)明(ming)显的(de)(de)(de)局限性(xing)(xing)(xing)(xing),其重复性(xing)(xing)(xing)(xing)差、由(you)(you)于(yu)(yu)聚集而引起的(de)(de)(de)粒径变(bian)化、颗(ke)粒与培养基(ji)组分(例如白蛋白)的(de)(de)(de)相互(hu)作(zuo)用(yong)(yong)(yong)以及颗(ke)粒被培养基(ji)溶(rong)解(Savi等人,2008年(nian)(nian)(nian);Fatisson等人,2012年(nian)(nian)(nian))。此(ci)外,吸入颗(ke)粒将(jiang)首先与肺(fei)表(biao)(biao)(biao)面活性(xing)(xing)(xing)(xing)剂相互(hu)作(zuo)用(yong)(yong)(yong),后(hou)者由(you)(you)上(shang)皮(pi)II型细胞产(chan)生,覆盖肺(fei)泡区域,以防(fang)止肺(fei)泡塌陷(xian)等功能。表(biao)(biao)(biao)面活性(xing)(xing)(xing)(xing)剂涂层可改(gai)变(bian)吸入颗(ke)粒的(de)(de)(de)表(biao)(biao)(biao)面特性(xing)(xing)(xing)(xing)和随后(hou)的(de)(de)(de)毒性(xing)(xing)(xing)(xing)。传统的(de)(de)(de)浸没(mei)(mei)式暴露(lu)法显然不具(ju)备模(mo)(mo)(mo)拟体(ti)内真(zhen)(zhen)实(shi)环(huan)境。相比(bi)之(zhi)下,气(qi)-液界面(ALI)模(mo)(mo)(mo)型中(zhong)(zhong),通过在(zai)气(qi)-界面处为(wei)上(shang)皮(pi)细胞暴露(lu)空气(qi)表(biao)(biao)(biao)面提供一层薄(bo)薄(bo)的(de)(de)(de)气(qi)道表(biao)(biao)(biao)面液体(ti)来真(zhen)(zhen)实(shi)反(fan)映体(ti)内环(huan)境(Jayaraman等人,2001年(nian)(nian)(nian))。此(ci)外,与浸没(mei)(mei)在(zai)培养基(ji)中(zhong)(zhong)生长(zhang)的(de)(de)(de)细胞相比(bi),气(qi)-液界面生长(zhang)的(de)(de)(de)上(shang)皮(pi)细胞具(ju)有(you)(you)很好的(de)(de)(de)分化结(jie)构和功能(Kameyama等人,2003年(nian)(nian)(nian))。因此(ci),ALI可以提供与动物(wu)(wu)吸入非常相似的(de)(de)(de)实(shi)验条件,尽管(guan)在(zai)其应用(yong)(yong)(yong)中(zhong)(zhong)仍(reng)有(you)(you)许多未解决的(de)(de)(de)问(wen)题,即在(zai)ALI或所(suo)谓的(de)(de)(de)体(ti)内表(biao)(biao)(biao)面活性(xing)(xing)(xing)(xing)剂中(zhong)(zhong)粒子(zi)与表(biao)(biao)(biao)面液体(ti)的(de)(de)(de)相互(hu)作(zuo)用(yong)(yong)(yong)动力(li)学、粒子(zi)的(de)(de)(de)理化变(bian)化以及对组成的(de)(de)(de)影响。本(ben)研(yan)(yan)究采(cai)用(yong)(yong)(yong)气(qi)液界面(ALI)暴露(lu)模(mo)(mo)(mo)型研(yan)(yan)究HSO3与SiO2纳米粒子(zi)协同肺(fei)毒性(xing)(xing)(xing)(xing),揭(jie)示复合空气(qi)污(wu)染物(wu)(wu)的(de)(de)(de)肺(fei)毒性(xing)(xing)(xing)(xing)及可能引起的(de)(de)(de)肺(fei)部**,特别是(shi)无机物(wu)(wu)和纳米颗(ke)粒复合污(wu)染物(wu)(wu)的(de)(de)(de)肺(fei)毒性(xing)(xing)(xing)(xing)具(ju)有(you)(you)重要(yao)意义(yi)。

文章摘要:

颗(ke)粒(li)物(wu)(PM)暴(bao)(bao)(bao)露对(dui)人体健康有(you)许多(duo)不(bu)(bu)利影响(xiang)。空(kong)气(qi)(qi)(qi)环境是一由各(ge)种复杂混合物(wu)构成(cheng)的(de)(de)(de)(de)(de)(de)(de)(de),然而,要厘清其(qi)中(zhong)单个污(wu)染(ran)(ran)(ran)物(wu)的(de)(de)(de)(de)(de)(de)(de)(de)毒性(xing)(xing)(xing)贡献是非常有(you)挑战性(xing)(xing)(xing)的(de)(de)(de)(de)(de)(de)(de)(de)。本研究的(de)(de)(de)(de)(de)(de)(de)(de)目(mu)(mu)的(de)(de)(de)(de)(de)(de)(de)(de)是使用二氧(yang)(yang)化(hua)(hua)硅纳(na)(na)米(mi)颗(ke)粒(li)(SiO2 NPs)和亚(ya)(ya)硫(liu)(liu)(liu)酸氢盐(yan)(yan)(yan)组成(cheng)的(de)(de)(de)(de)(de)(de)(de)(de)气(qi)(qi)(qi)溶(rong)(rong)胶作(zuo)为模拟粒(li)子(zi)相关的(de)(de)(de)(de)(de)(de)(de)(de)高硫(liu)(liu)(liu)污(wu)染(ran)(ran)(ran)物(wu)。然后(hou),使用气(qi)(qi)(qi)-液(ye)界(jie)面(mian)(mian)(ALI)的(de)(de)(de)(de)(de)(de)(de)(de)暴(bao)(bao)(bao)露室在(zai)(zai)细(xi)胞(bao)水(shui)平(ping)上评估二氧(yang)(yang)化(hua)(hua)硫(liu)(liu)(liu)对(dui)健康的(de)(de)(de)(de)(de)(de)(de)(de)影响(xiang)。BEAS-2B细(xi)胞(bao)在(zai)(zai)气(qi)(qi)(qi)-液(ye)界(jie)面(mian)(mian)(ALI)分别暴(bao)(bao)(bao)露于(yu)纳(na)(na)米(mi)二氧(yang)(yang)化(hua)(hua)硅、亚(ya)(ya)硫(liu)(liu)(liu)酸氢盐(yan)(yan)(yan)气(qi)(qi)(qi)溶(rong)(rong)胶以及涂(tu)(tu)有(you)亚(ya)(ya)硫(liu)(liu)(liu)酸氢盐(yan)(yan)(yan)的(de)(de)(de)(de)(de)(de)(de)(de)二氧(yang)(yang)化(hua)(hua)硅(SiO2-NH2@HSO3)气(qi)(qi)(qi)溶(rong)(rong)胶中(zhong)3 小时。根据暴(bao)(bao)(bao)露剂(ji)量(liang)比较其(qi)对(dui)细(xi)胞(bao)的(de)(de)(de)(de)(de)(de)(de)(de)毒性(xing)(xing)(xing)。二氧(yang)(yang)化(hua)(hua)硅纳(na)(na)米(mi)颗(ke)粒(li)(SiO2NPs)单独(du)暴(bao)(bao)(bao)露于(yu)空(kong)气(qi)(qi)(qi)中(zhong)不(bu)(bu)产(chan)生明显(xian)(xian)的(de)(de)(de)(de)(de)(de)(de)(de)细(xi)胞(bao)毒性(xing)(xing)(xing),但涂(tu)(tu)有(you)亚(ya)(ya)硫(liu)(liu)(liu)酸氢盐(yan)(yan)(yan)的(de)(de)(de)(de)(de)(de)(de)(de)二氧(yang)(yang)化(hua)(hua)硅(SiO2-NH2@HSO3)气(qi)(qi)(qi)溶(rong)(rong)胶暴(bao)(bao)(bao)露显(xian)(xian)著(zhu)降低了(le)细(xi)胞(bao)活性(xing)(xing)(xing),并以剂(ji)量(liang)依赖(lai)性(xing)(xing)(xing)方式(shi)增强了(le)细(xi)胞(bao)活性(xing)(xing)(xing)氧(yang)(yang)的(de)(de)(de)(de)(de)(de)(de)(de)产(chan)生。因此(ci),过(guo)度(du)的(de)(de)(de)(de)(de)(de)(de)(de)氧(yang)(yang)化(hua)(hua)应激导致线(xian)粒(li)体损(sun)伤和细(xi)胞(bao)凋亡。气(qi)(qi)(qi)-液(ye)界(jie)面(mian)(mian)暴(bao)(bao)(bao)露方法(fa)可能(neng)反映人体呼吸系(xi)统的(de)(de)(de)(de)(de)(de)(de)(de)实(shi)际生理暴(bao)(bao)(bao)露状况(kuang)。硫(liu)(liu)(liu)酸盐(yan)(yan)(yan)作(zuo)为空(kong)气(qi)(qi)(qi)污(wu)染(ran)(ran)(ran)二氧(yang)(yang)化(hua)(hua)硫(liu)(liu)(liu)的(de)(de)(de)(de)(de)(de)(de)(de)衍(yan)生物(wu),加剧了(le)可吸入PMS的(de)(de)(de)(de)(de)(de)(de)(de)毒性(xing)(xing)(xing)作(zuo)用。这(zhei)一结果可能(neng)是由于(yu)纳(na)(na)米(mi)颗(ke)粒(li)具有(you)大表面(mian)(mian)积的(de)(de)(de)(de)(de)(de)(de)(de)特性(xing)(xing)(xing),在(zai)(zai)气(qi)(qi)(qi)溶(rong)(rong)胶暴(bao)(bao)(bao)露期间能(neng)够携带更多(duo)的(de)(de)(de)(de)(de)(de)(de)(de)亚(ya)(ya)硫(liu)(liu)(liu)酸盐(yan)(yan)(yan)并到达靶细(xi)胞(bao)。硫(liu)(liu)(liu)酸盐(yan)(yan)(yan)水(shui)平(ping)为目(mu)(mu)前的(de)(de)(de)(de)(de)(de)(de)(de)PM2.5空(kong)气(qi)(qi)(qi)污(wu)染(ran)(ran)(ran)指数提(ti)供(gong)了(le)一个有(you)意义的(de)(de)(de)(de)(de)(de)(de)(de)补(bu)充,以实(shi)现(xian)更好的(de)(de)(de)(de)(de)(de)(de)(de)人类健康防(fang)护。

实验重要(yao)数据

本(ben)研究所有ALI暴露(lu)实验(yan)均在德(de)国CLUTEX RFS COMPACT系统中完成,RFS COMPACT系统内(nei)可(ke)同时(shi)进行(xing)对(dui)照组(zu)与实验(yan)组(zu)两组(zu)实验(yan)。

 The exposure doses of NM aerosols. The size distributionsof aerosols of (A) SiO2-OH, (C) SiO2-NH2 and (E) SiO2-NH2@HSO3 in the 14.6e661.2 nm particle size range. The mass concentrations ofaerosol particles of (B) SiO2-OH, (D) SiO2-NH2 and (F) SiO2-NH2@HSO3 in the 14.6e661.2 nm particle size range after exposure on BEAS-2Bcells for 3 h at ALI.

Cellular toxicitiesdetected in BEAS-2B after ALI exposure to various PM aerosols. Cell viability(A) and cell membrane integrity analysis (B) after 3 h exposure. C1 is the ALI exposurecontrol of filteredclean air; C2 is the normal cell control cultured in incubator. The ROS levels(C) and ATP production (D) in BEAS-2B cells after ALI exposure for 3 h and withan extra incubation for 5 h. N =3,Control group is the ALI exposure of filtered clean air. The data represents onecandidate experiment result from at least three independent experiments. *P < 0.05, #P <0.01 vs the indicated groups.

The live & dead testing results of BEAS-2B cells after exposureto aerosols for 3 h at ALI and extra incubation in ALI chamber for 5 h. Thedata represent the mean of at least three independent experiments normalized tocontrols of ALI. C1 is the ALI exposure control of filtered clean air; C2 is the normal cell controlcultured in incubator. *P <0.05, #P < 0.01 vs the indicated groups.

 Expression levels of marker genes in BEAS-2Bcells after ALI exposure to different PM aerosols for 3 h and with an extraincubation for 5 h. (A) Bax; (B) Bcl-2;(C) Bcl-2/Baxratio; (D) Caspase-3; (E) Chop; and (F) Xbp-1s.The data represents one candidate experiment result from at least threeindependent experiments. N =5,Control group is the ALI exposure of clean air. *P < 0.05, #P <0.01 vs the indicated groups.

恭喜!国家纳米中(zhong)心使(shi)用CULTEX细胞暴露技(ji)(ji)术研(yan)究空(kong)气污染的(de)高分文章发表。该(gai)研(yan)究得到国家科(ke)(ke)技(ji)(ji)部(bu)、国家自然科(ke)(ke)学(xue)基金(jin)、国家杰出青(qing)年科(ke)(ke)学(xue)基金(jin)和(he)中(zhong)国科(ke)(ke)学(xue)院前(qian)沿科(ke)(ke)学(xue)重点研(yan)究计(ji)划项目的(de)资助。