CN114487158A - 一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法 - Google Patents
一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法 Download PDFInfo
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Abstract
本发明提供一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,属于雾霾颗粒物毒性评价领域。该方法以PM2.5为基础,采用金属螯合、有机物萃取和ROS消耗的方式来去除PM2.5中的金属,PAHs和ROS,这些被去除金属、PAHs和ROS后的PM2.5对巨噬细胞NLRP3炎性体激活及小鼠肺部纤维化形成影响,本发明通过对雾霾颗粒物的组分改造前后毒性水平的比较,鉴定了PM2.5引发肺部慢性纤维化的来源。
Description
技术领域
本发明涉及雾霾颗粒物(PM2.5)毒性评价领域,具体涉及一种通过去除雾霾颗粒物中金属离子、多环芳烃及活性氧自由基组分来鉴定PM2.5引发肺部慢性纤维化毒性来源的方法。
背景技术
随着全球城市化和工业化的加速发展,空气污染日益严重,其对人类健康的负面影响逐渐引起广泛关注。越来越多的毒理学、流行病学等相关领域的研究表明,细颗粒物(PM2.5)对呼吸系统的危害存在浓度依赖关系。有报道称PM2.5浓度每增加10μg m-3/d,呼吸道疾病患病率增加2.07%,住院率增加8%。越来越多的证据表明,PM2.5暴露引起的炎症是肺部疾病发展的主要驱动因素,如哮喘、肺炎症或纤维化、肺癌等。其中,NLRP3炎性体激活和IL-1β的释放是PM2.5引发肺部炎症的重要因素。过去研究报道,PM2.5组成中的可溶的化学和生物成分,包括硝酸盐、硫酸盐、铵、金属、多芳香烃(PAHs)、细菌内毒素、过敏原和表面活性氧(ROS)等在毒性诱导方面发挥不可忽略的作用。因此,开发有效的鉴定方法来确定PM2.5引发毒性的组分来源对于快速确定环境污染源有着重要意义。
发明内容
本发明提供一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,本发明通过去除PM2.5的金属离子、PAHs及ROS,评价其对于肺部慢性纤维化的影响,鉴定PM2.5的个别组分是否为其毒性来源。
为了实现上述目的,本发明的技术方案具体如下:
一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,包括以下步骤:
步骤一:获取PM2.5颗粒;
步骤二:将步骤一的PM2.5颗粒分别进行金属元素、PAHs和ROS的定量分析;
步骤三:对PM2.5颗粒中的金属、PAHs或ROS进行去除,分别得到MetalPM2.5、PAHPM2.5或ROSPM2.5;
步骤四:用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,采用Elisa法检测IL-1β的释放,鉴定炎性体的激活状态;
步骤五:通过口咽法使得小鼠吸入MetalPM2.5、PAHPM2.5或ROSPM2.5,并进行Massion染色,鉴定改性后的雾霾颗粒物对肺部纤维化的影响。
优选的是,所述的步骤三中PM2.5颗粒中金属组分的去除,具体为:
采用Chelex 100树脂固相萃取(SPE)柱吸附PM2.5中的重金属,Chelex 100树脂经过超纯水预处理后,在聚丙烯SPE储层中加入Chelex 100树脂,制备Chelex 100色谱柱,然后,将PM2.5水悬浮液通过色谱柱,收集通过的溶液,冷冻干燥保存,该样品被命名为MetalPM2.5。
优选的是,所述的聚丙烯SPE储层的体积mL:Chelex 100树脂的质量g:PM2.5水悬浮液的体积mL为3:0.2:1。
优选的是,所述的PM2.5水悬浮液的浓度为5mg mL-1。
优选的是,所述的步骤三中PM2.5颗粒中PAHs的去除,具体为:
将PM2.5悬浮液与有机溶剂混合物混合提取有机物,所得混合物经离心,将沉淀物冻干保存,该样品命名为PAHPM2.5,所述的有机溶剂混合物为正己烷、二氯甲烷和甲醇的混合。
优选的是,所述的PM2.5水悬浮液的浓度为5mg mL-1。
优选的是,所述的正己烷、二氯甲烷和甲醇体积比为1:1:1。
优选的是,所述的步骤三中PM2.5颗粒中ROS的去除,具体为:
将PM2.5水悬浮液与抗坏血酸混合,所得混合物经离心,以去除多余的AA,将沉淀冻干,保存,该样本被命名为ROSPM2.5。
优选的是,所述的PM2.5水悬浮液的浓度为1mg mL-1,抗坏血酸的浓度为250μg mL-1。
优选的是,所述的步骤四具体为:
步骤1、细胞培养:
THP-1细胞培养在包含10%的胎牛血清、100U mL-1的青霉素和100mg mL-1链霉素的RPMI1640培养基中,其培养条件是37℃和5%的CO2且每隔一天更换培养基一次;
步骤2、Elisa检测IL-1β的释放:
将THP-1细胞以每孔3×104个细胞的密度,置于96孔板中,用100μL细胞培养基孵育16h,培养基中含有1μg mL-1佛波尔12-肉豆蔻酸13-乙酸酯(PMA)诱导THP-1细胞分化,然后,用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,在脂多糖(LPS,10ng mL-1)的存在下再处理24小时,收集细胞上清,采用Elisa法检测IL-1β。
优选的是,所述的步骤五具体为:
将8周龄雌性Balb/c小鼠在标准的实验室条件下培养,动物暴露于PM2.5是通过NIOSH描述的口咽抽吸法进行的,50μL PBS包含400μg mL-1PM2.5、MetalPM2.5、PAHPM2.5或ROSPM2.5被滴在小鼠的舌头后方,捏住鼻腔让其进入肺部,21天后处死小鼠,并收集肺组织,进行Masson染色。
本发明的有益效果
本发明提供一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,该方法以PM2.5为基础,采用金属螯合、有机物萃取和ROS消耗的方式来去除PM2.5中的金属,PAHs和ROS,本发明通过阳离子交换树脂(Chelex 100)来去除主要的过渡金属,Chelex 100对金属离子具有较高的结合强度,并具有较高的去除特异性,并采用有机溶剂(正己烷、二氯甲烷和甲醇)的多重萃取工艺去除PM2.5中的PAHs。此外,抗坏血酸是一种水溶性抗氧化分子,在解毒和中和ROS的循环通路中起主要底物作用,本发明通过抗坏血酸对PM2.5进行处理,去除ROS。这些被去除金属、PAHs和ROS后的PM2.5对巨噬细胞(如THP-1细胞)NLRP3炎性体激活及小鼠肺部纤维化形成影响,由此鉴定组分是否为肺部慢性纤维化的来源。PM2.5毒性组分来源的鉴定对于确定环境污染源和未来环境修复有着重要的意义。
附图说明
下面结合附图和具体实施方式对本发明作进一步详细说明。
图1为PM2.5的组成图;
图2为金属螯合前后PM2.5中各金属元素的含量柱状图;
图3为有机溶剂萃取前后PM2.5中PAHs的含量柱状图;
图4为AA处理前后PM2.5中ROS的变化曲线图;(基于DCF荧光强度)。
图5为PM2.5和MetalPM2.5所诱导的IL-1β的产生柱状图;
图6为PM2.5和PAHPM2.5所诱导的IL-1β的产生柱状图;
图7为PM2.5和ROSPM2.5所诱导的IL-1β的产生柱状图;
图8为PM2.5、MetalPM2.5、PAHPM2.5和ROSPM2.5所诱导的小鼠肺部纤维化的形成照片。
具体实施方式
一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,包括以下步骤:
步骤一:获取PM2.5颗粒;
步骤二:将步骤一的PM2.5颗粒分别进行金属元素、PAHs和ROS的定量分析;
步骤三:对PM2.5颗粒中的金属、PAHs或ROS进行去除,分别得到MetalPM2.5、PAHPM2.5或ROSPM2.5;
步骤四:用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,采用Elisa法检测IL-1β的释放,鉴定炎性体的激活状态;
步骤五:通过口咽法使得小鼠吸入MetalPM2.5、PAHPM2.5或ROSPM2.5,并进行Massion染色,鉴定改性后的雾霾颗粒物对肺部纤维化的影响。
按照本发明,所述的PM2.5颗粒的获取,具体为:采用Anderson G1200采样器在硝酸纤维素过滤膜上以16.7L min-1的流速采集PM2.5,采样器设置在约20m高的建筑屋顶上,通过刮刀小心地从滤膜上刮取PM2.5颗粒。
按照本发明,所述的步骤二中PM2.5颗粒进行金属元素,具体包括:
采样前后的重量之差即为PM2.5颗粒总质量,在恒温(20±2℃)和相对湿度(40±4%)下稳定24h后,用电子天平测得PM2.5质量浓度,检出限为1μg,每个样品用20-50mLHNO3/HF/HClO4酸性混合物(体积比4:4:1)在200℃下处理24h,使样品分解,干燥后的消化液用1%的HNO3溶液稀释至100mL,采用电感耦合等离子体质谱仪(ICP-MS)对其进行分析。采用ICP-MS对样品进行金属元素的定量分析,所述的金属元素为Al、Mg、Zn、Pb、V、Mn、Fe、Co、Ni、Cu、Hg、Ge、Ba、Ce。
按照本发明,所述的步骤二中PM2.5颗粒进行PAHs分析,具体包括:
将PM2.5溶于二氯甲烷中2h,悬浮液10000rpm离心0.5h,真空下旋转蒸发浓缩上清,采用安捷伦7890气相色谱、5975C质谱检测器和DB-5毛细管柱(30m×0.25mm×0.25μm)测定多环芳烃(PAHs)含量。
按照本发明,所述的步骤二中PM2.5颗粒进行ROS分析,具体包括:
用2',7'-二氯二氢荧光素二乙酸酯(H2DCFDA)荧光检测总ROS水平,将50μgH2DCFDA与17.3μL乙醇混合,再加入692μL 0.01mol L-1氢氧化钠溶液,然后,加入3500μL磷酸钠缓冲液(25mmol L-1,pH=7.4)孵育30min,形成29μmol L-1DCF溶液,在96孔板的每个孔中加入80μL DCF工作液和一定量颗粒物悬液,孵育2h,用SpectraMax M3酶标仪在500–600nm的范围,490nm激发波长下检测其发射光谱。
按照本发明,所述的步骤三中PM2.5颗粒中金属组分的去除,具体为:
采用Chelex 100树脂固相萃取(SPE)柱吸附PM2.5中的重金属,Chelex 100树脂经过超纯水预处理后,在聚丙烯SPE储层中加入Chelex 100树脂,制备Chelex 100色谱柱,然后,将PM2.5水悬浮液通过色谱柱,收集通过的溶液,冷冻干燥保存,该样品被命名为MetalPM2.5。所述的聚丙烯SPE储层的体积mL:Chelex 100树脂的质量g:PM2.5水悬浮液的体积mL优选为3:0.2:1;所述的PM2.5水悬浮液的浓度优选为5mg mL-1;所述的PM2.5水悬浮液通过色谱柱的流速优选为0.2mL min-1。
按照本发明,所述的步骤三中PM2.5颗粒中PAHs的去除,具体为:
将PM2.5悬浮液与有机溶剂混合物混合提取有机物,所述的混合时间优选为1-2h,所得混合物经离心,所述的混合物离心速率优选为10000-12000rpm,时间优选为20-60min,将沉淀物冻干保存,该样品命名为PAHPM2.5,所述的有机溶剂混合物为正己烷、二氯甲烷和甲醇的混合,所述的正己烷、二氯甲烷和甲醇体积比优选为1:1:1;所述的PM2.5水悬浮液的浓度优选为5mg mL-1;PM2.5悬浮液与有机溶剂混合物的体积比优选为1:1。
按照本发明,所述的步骤三中PM2.5颗粒中ROS的去除,具体为:
将PM2.5水悬浮液与抗坏血酸混合,所述的混合时间优选为20-60min,所得混合物经离心,所述的混合物离心速率优选为10000-20000rpm,时间优选为15-30min,以去除多余的AA,将沉淀冻干,保存,该样本被命名为ROSPM2.5。所述的PM2.5水悬浮液的浓度优选为1mgmL-1,抗坏血酸的浓度优选为250μg mL-1;所述的PM2.5水悬浮液10mL与250μg mL-1抗坏血酸的体积比优选为1:4。
按照本发明,所述的步骤四具体为:
步骤1、细胞培养:
THP-1细胞培养在包含10%的胎牛血清、100U mL-1的青霉素和100mg mL-1链霉素的RPMI1640培养基中,其培养条件是37℃和5%的CO2且每隔一天更换培养基一次;
步骤2、Elisa检测IL-1β的释放:
将THP-1细胞以每孔3×104个细胞的密度,置于96孔板中,用100μL细胞培养基孵育16h,培养基中含有1μg mL-1佛波尔12-肉豆蔻酸13-乙酸酯(PMA)诱导THP-1细胞分化,然后,用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,在脂多糖(LPS,10ng mL-1)的存在下再处理24小时,收集细胞上清,采用Elisa法检测IL-1β。
按照本发明,所述的步骤五具体为:
将8周龄雌性Balb/c小鼠在标准的实验室条件下培养,动物暴露于PM2.5是通过NIOSH描述的口咽抽吸法进行的,50μL PBS包含400μg mL-1PM2.5、MetalPM2.5、PAHPM2.5或ROSPM2.5被滴在小鼠的舌头后方,捏住鼻腔让其进入肺部,21天后处死小鼠,并收集肺组织,进行Masson染色。
下面结合附图和具体实施例对本发明做详细说明。
实施例1
一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,具体步骤如下:
1.PM2.5的获取:
在中国长春,采用Anderson G1200采样器在硝酸纤维素过滤膜上以16.7L min-1的流速采集PM2.5。采样器设置在约20m高的建筑屋顶上,该建筑被认为是一个集住宅、交通、建筑和工业为一体的代表性区域。通过刮刀小心地从滤膜上刮取PM2.5颗粒。
2.金属分析
采样前后的重量之差即为PM2.5颗粒总质量。PM2.5质量浓度的不确定度估计为总测量值的5%。在恒温(20±2℃)和相对湿度(40±4%)下稳定24h后,用电子天平测得PM2.5质量浓度,检出限为1μg。每个样品用20-50mL HNO3/HF/HClO4酸性混合物(体积比4:4:1)在200℃下处理24h,使样品分解。干燥后的消化液用1%的HNO3溶液稀释至100mL。采用电感耦合等离子体质谱仪(ICP-MS)对其进行分析。采用ICP-MS对样品进行金属元素(Al、Mg、Zn、Pb、V、Mn、Fe、Co、Ni、Cu、Hg、Ge、Ba、Ce)的定量分析如图1。
3.PAHs分析
将PM2.5溶于二氯甲烷中2h,悬浮液10000rpm离心0.5h,真空下旋转蒸发浓缩上清。采用安捷伦7890气相色谱、5975C质谱检测器和DB-5毛细管柱(30m×0.25mm×0.25μm)测定多环芳烃(PAHs)含量如图1。
4、ROS分析
用2',7'-二氯二氢荧光素二乙酸酯(H2DCFDA)荧光检测总ROS水平,将50μgH2DCFDA与17.3μL乙醇混合,再加入692μL 0.01mol L-1氢氧化钠溶液,然后,加入3500μL磷酸钠缓冲液(25mmol L-1,pH=7.4)孵育30min,形成29μmol L-1DCF溶液,在96孔板的每个孔中加入80μL DCF工作液和一定量颗粒物悬液,孵育2h,用SpectraMax M3酶标仪在500–600nm的范围,490nm激发波长下检测其发射光谱。
5、PM2.5中金属组分的去除
采用Chelex 100树脂固相萃取(SPE)柱吸附PM2.5中的重金属。Chelex 100树脂经过超纯水预处理后,在3mL聚丙烯SPE储层中加入0.2g Chelex 100树脂,制备Chelex 100色谱柱。然后,将1mL 5mg mL-1PM2.5水悬浮液以0.2mL min-1的流速通过色谱柱。收集通过的溶液,在4℃下冷冻干燥保存。该样品被命名为MetalPM2.5如图2。
6、PM2.5中PAHs组分的去除
将3mL 5mg mL-1PM2.5悬浮液与3mL有机溶剂混合物(1mL正己烷、1mL二氯甲烷和1mL甲醇)混合2h提取有机物。所得混合物在12000rpm离心20min,沉淀物在4℃冻干保存。该样品命名为PAHPM2.5如图3。
7、PM2.5表面ROS的去除
将1mg mL-1PM2.5水悬浮液10mL与250μg mL-1抗坏血酸(AA)40mL混合20min。所得混合物以10000转/分钟离心15分钟,以去除多余的AA。将沉淀冻干,4℃保存。这个样本被命名为ROSPM2.5如图4。
8、细胞培养
THP-1细胞培养在包含10%的胎牛血清、100U mL-1的青霉素和100mg mL-1链霉素的RPMI1640培养基中。其培养条件是37℃和5%的CO2且每隔一天更换培养基一次。
9、Elisa检测IL-1β的释放:
将THP-1细胞以每孔3×104个细胞的密度,置于96孔板中,用100μL细胞培养基孵育16h。培养基中含有1μg mL-1佛波尔12-肉豆蔻酸13-乙酸酯(PMA)诱导THP-1细胞分化。然后,用MetalPM2.5、PAHPM2.5、ROSPM2.5或PM2.5处理不同分化的THP-1细胞,在脂多糖(LPS,10ng mL-1)的存在下再处理24小时。收集细胞上清,采用Elisa法检测IL-1β。如图5、图6和图7所示,当显著性差异p<0.05时,被认为显著降低了PM2.5的炎性体激活能力,图5-7说明,巨噬细胞经MetalPM2.5、PAHPM2.5或ROSPM2.5孵育后释放白介素1β的水平显著低于经PM2.5孵育后释放的水平(p<0.05)。
10、Masson染色检测小鼠肺部纤维化的形成:
8周龄雌性Balb/c小鼠购于北京维通利华动物科技有限公司,所有的动物都在标准的实验室条件下培养。动物暴露于PM2.5是通过NIOSH描述的口咽抽吸法进行的,50μL PBS包含400μg mL-1PM2.5、MetalPM2.5、PAHPM2.5或ROSPM2.5被滴在小鼠的舌头后方,捏住鼻腔让其进入肺部。21天后处死小鼠,收集肺组织,进行Masson染色。如图8所示,小鼠经MetalPM2.5、PAHPM2.5或ROSPM2.5暴露后的肺部纤维化水平被认定为窦周纤维化,而原始的PM2.5组引发的纤维化水平被认定为门静脉窦周纤维化。
Claims (10)
1.一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,包括以下步骤:
步骤一:获取PM2.5颗粒;
步骤二:将步骤一的PM2.5颗粒分别进行金属元素、PAHs和ROS的定量分析;
步骤三:对PM2.5颗粒中的金属、PAHs或ROS进行去除,分别得到MetalPM2.5、PAHPM2.5或ROSPM2.5;
步骤四:用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,采用Elisa法检测IL-1β的释放,鉴定炎性体的激活状态;
步骤五:通过口咽法使得小鼠吸入MetalPM2.5、PAHPM2.5或ROSPM2.5,并进行Massion染色,鉴定改性后的雾霾颗粒物对肺部纤维化的影响。
2.根据权利要求1所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的步骤三中PM2.5颗粒中金属组分的去除,具体为:
采用Chelex 100树脂固相萃取柱吸附PM2.5中的重金属,Chelex 100树脂经过超纯水预处理后,在聚丙烯SPE储层中加入Chelex 100树脂,制备Chelex 100色谱柱,然后,将PM2.5水悬浮液通过色谱柱,收集通过的溶液,冷冻干燥保存,该样品被命名为MetalPM2.5。
3.根据权利要求2所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的聚丙烯SPE储层的体积mL:Chelex 100树脂的质量g:PM2.5水悬浮液的体积mL为3:0.2:1。
4.根据权利要求1所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的步骤三中PM2.5颗粒中PAHs的去除,具体为:
将PM2.5悬浮液与有机溶剂混合物混合提取有机物,所得混合物经离心,将沉淀物冻干保存,该样品命名为PAHPM2.5,所述的有机溶剂混合物为正己烷、二氯甲烷和甲醇的混合。
5.根据权利要求2或4所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的PM2.5水悬浮液的浓度为5mg mL-1。
6.根据权利要求4所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的正己烷、二氯甲烷和甲醇体积比为1:1:1。
7.根据权利要求1所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的步骤三中PM2.5颗粒中ROS的去除,具体为:
将PM2.5水悬浮液与抗坏血酸混合,所得混合物经离心,以去除多余的AA,将沉淀冻干,保存,该样本被命名为ROSPM2.5。
8.根据权利要求7所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的PM2.5水悬浮液的浓度为1mg mL-1,抗坏血酸的浓度为250μg mL-1。
9.根据权利要求1所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的步骤四具体为:
步骤1、细胞培养:
THP-1细胞培养在包含10%的胎牛血清、100U mL-1的青霉素和100mg mL-1链霉素的RPMI1640培养基中,其培养条件是37℃和5%的CO2且每隔一天更换培养基一次;
步骤2、Elisa检测IL-1β的释放:
将THP-1细胞以每孔3×104个细胞的密度,置于96孔板中,用100μL细胞培养基孵育16h,培养基中含有1μg mL-1佛波尔12-肉豆蔻酸13-乙酸酯诱导THP-1细胞分化,然后,用MetalPM2.5、PAHPM2.5或ROSPM2.5处理不同分化的THP-1细胞,在脂多糖LPS,10ng mL-1的存在下再处理24小时,收集细胞上清,采用Elisa法检测IL-1β。
10.根据权利要求1所述的一种鉴定改性后的雾霾颗粒物毒性及对肺部纤维化影响的方法,其特征在于,所述的步骤五具体为:
将8周龄雌性Balb/c小鼠在标准的实验室条件下培养,动物暴露于PM2.5是通过NIOSH描述的口咽抽吸法进行的,50μL PBS包含400μg mL-1PM2.5、MetalPM2.5、PAHPM2.5或ROSPM2.5被滴在小鼠的舌头后方,捏住鼻腔让其进入肺部,21天后处死小鼠,并收集肺组织,进行Masson染色。
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Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110004414A1 (en) * | 2009-06-06 | 2011-01-06 | Ceetox, Inc. | Method for Predicting Respiratory Toxicity of Compounds |
US20110152338A1 (en) * | 2008-06-24 | 2011-06-23 | Amira Pharmaceuticals, Inc. | Cycloalkane[b]indole antagonists of prostaglandin d2 receptors |
US20150055127A1 (en) * | 2012-04-02 | 2015-02-26 | Asml Netherlands B.V. | Particulate Contamination Measurement Method and Apparatus |
CN106823562A (zh) * | 2016-12-27 | 2017-06-13 | 上海纳米技术及应用国家工程研究中心有限公司 | 一种人工肺表面活性物质用于去除pm2.5的方法 |
CN106841440A (zh) * | 2017-01-20 | 2017-06-13 | 山东省分析测试中心 | 一种环境中有机酸的检测方法 |
US20170165591A1 (en) * | 2015-12-14 | 2017-06-15 | Waters Technologies Corporation | Chromatography filter |
CN110736791A (zh) * | 2018-07-20 | 2020-01-31 | 中国科学院大连化学物理研究所 | 一种雾霾相关多环芳烃毒性评价方法 |
KR20200110019A (ko) * | 2019-03-15 | 2020-09-23 | 영남대학교 산학협력단 | 2,2'-바이피리딘 화합물을 포함하는 호흡기 질환 예방 또는 치료용 조성물 |
CN114487432A (zh) * | 2021-12-27 | 2022-05-13 | 中国科学院长春应用化学研究所 | 一种鉴定改变物理化性质后的雾霾颗粒毒性及对肺部纤维化影响的方法 |
-
2021
- 2021-12-27 CN CN202111610368.6A patent/CN114487158A/zh active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110152338A1 (en) * | 2008-06-24 | 2011-06-23 | Amira Pharmaceuticals, Inc. | Cycloalkane[b]indole antagonists of prostaglandin d2 receptors |
US20110004414A1 (en) * | 2009-06-06 | 2011-01-06 | Ceetox, Inc. | Method for Predicting Respiratory Toxicity of Compounds |
US20150055127A1 (en) * | 2012-04-02 | 2015-02-26 | Asml Netherlands B.V. | Particulate Contamination Measurement Method and Apparatus |
US20170165591A1 (en) * | 2015-12-14 | 2017-06-15 | Waters Technologies Corporation | Chromatography filter |
CN106823562A (zh) * | 2016-12-27 | 2017-06-13 | 上海纳米技术及应用国家工程研究中心有限公司 | 一种人工肺表面活性物质用于去除pm2.5的方法 |
CN106841440A (zh) * | 2017-01-20 | 2017-06-13 | 山东省分析测试中心 | 一种环境中有机酸的检测方法 |
CN110736791A (zh) * | 2018-07-20 | 2020-01-31 | 中国科学院大连化学物理研究所 | 一种雾霾相关多环芳烃毒性评价方法 |
KR20200110019A (ko) * | 2019-03-15 | 2020-09-23 | 영남대학교 산학협력단 | 2,2'-바이피리딘 화합물을 포함하는 호흡기 질환 예방 또는 치료용 조성물 |
CN114487432A (zh) * | 2021-12-27 | 2022-05-13 | 中国科学院长春应用化学研究所 | 一种鉴定改变物理化性质后的雾霾颗粒毒性及对肺部纤维化影响的方法 |
Non-Patent Citations (15)
Title |
---|
A. DOLATABADIAN ET AL.: "The Effects of Foliar Application of Ascorbic Acid (Vitamin C) on Antioxidant Enzymes Activities, Lipid Peroxidation and Proline Accumulation of Canola (Brassica napus L.) under Conditions of Salt Stress", 《JOURNAL OF AGRONOMY AND CROP SCIENCE》, vol. 194, no. 3, 5 March 2008 (2008-03-05), pages 206 - 213 * |
BRIAN BOURGEOIS ET AL.: "The influence of Hurricanes Katrina and Rita on the inflammatory cytokine response and protein expression in A549 cells exposed to PM2.5 collected in the Baton Rouge–Port Allen industrial corridor of Southeastern Louisiana in 2005", 《TOXICOLOGY MECHANISMS AND METHODS》, vol. 24, no. 3, 30 January 2014 (2014-01-30), pages 220 - 242 * |
CAO WEI ET AL.: "NLRP3 inflammasome activation determines the fibrogenic potential of PM2.5 air pollution particles in the lung", 《JOURNAL OF ENVIRONMENTAL SCIENCES》, vol. 111, 15 May 2021 (2021-05-15), pages 429 - 441 * |
HUI JIA ET AL.: "PM2.5-induced pulmonary inflammation via activating of the NLRP3/caspase-1 signaling pathway", 《ENVIRON TOXICOL》, vol. 36, no. 3, 31 March 2021 (2021-03-31), pages 298 - 307 * |
KUKKA AIMONEN ET AL.: "Effect of Surface Modification on the Pulmonary and Systemic Toxicity of Cellulose Nanofibrils", 《BIOMACROMOLECULES》, vol. 23, no. 7, 9 June 2022 (2022-06-09), pages 2752 - 2766 * |
RUNXIAO ZHENG ET AL.: "Property-activity relationship between physicochemical properties of PM2.5 and their activation of NLRP3 inflammasome", 《NANOIMPACT》, vol. 25, 11 January 2022 (2022-01-11), pages 1 - 10 * |
WANG, BAOMING ET AL.: "Maternal Particulate Matter Exposure Impairs Lung Health and Is Associated with Mitochondrial Damage", 《ANTIOXIDANTS》, vol. 10, no. 7, 31 July 2021 (2021-07-31), pages 1029 * |
YAN, JUNYAN ET AL.: "Industrial PM2.5 cause pulmonary adverse effect through RhoA/ROCK pathway", 《SCIENCE OF THE TOTAL ENVIRONMENT》, vol. 599, 1 December 2017 (2017-12-01), pages 1658 - 1666, XP085071591, DOI: 10.1016/j.scitotenv.2017.05.107 * |
丽娜・马达尼亚孜;叶晓芳;王佳佳;李国星;潘小川;: "非沙尘天气大气颗粒物中金属元素浓度与儿童肺功能相关性的初步研究", 《环境与健康杂志》, vol. 28, no. 07, 20 July 2011 (2011-07-20), pages 573 - 575 * |
吕烨;徐珊珊;刘卫艳;张文辉;: "PM_(2.5)组分的细胞毒性效应研究进展", 《中国预防医学杂志》, vol. 19, no. 04, 10 April 2018 (2018-04-10), pages 313 - 315 * |
李怡 等: "沙尘细颗粒物吸入对大鼠肺组织TNF-α、MMP、TGF-β1及部分胶原蛋白等指标影响的研究", 《中国呼吸与危重监护杂志》, vol. 19, no. 3, 31 March 2020 (2020-03-31), pages 264 - 269 * |
梁峰;石莹;张萱;刘忠英;刘志强;: "电离辐射诱导T淋巴细胞的差异蛋白质组学研究", 《高等学校化学学报》, vol. 31, no. 06, 10 June 2010 (2010-06-10), pages 1143 - 1147 * |
熊丽林 等: "PM2.5诱导的炎性反应对心血管内皮的损伤及分子机制研究", 《中国博士学位论文全文数据库 医药卫生科技辑》, no. 1, 15 January 2021 (2021-01-15), pages 055 - 21 * |
贾玉巧;赵晓红;郭新彪;: "大气颗粒物PM10和PM2.5对人肺成纤维细胞及其炎性因子分泌的影响", 《环境与健康杂志》, vol. 28, no. 03, 20 March 2011 (2011-03-20), pages 206 - 208 * |
郑润晓: "纳米颗粒物基于NLRP3炎性体激活而引发的慢性疾病研究及其在肿瘤治疗中的应用", 《中国博士学位论文全文数据库 工程科技Ⅰ辑》, no. 1, 15 January 2023 (2023-01-15), pages 020 - 232 * |
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