大气混合污染物对大鼠血清及肺组织CC16表达的影响
摘要
目的目前城市大气污染问题已经严重威胁着人类的健康,为了探索大气污染物对肺组织损伤的毒性作用机制,我们通过大气混合污染物的染尘染毒实验,了解大气混合污染物作用后的大鼠血清及肺组织CC16的表达水平,为预防大气污染对人体的损害提供依据。
方法本研究在乙醚麻醉下,通过将实验组大鼠注入含有PM2.5的生理盐水混悬液和吸入含有SO2、NO2、CO的空气混合气来建立动物染尘染毒模型。应用ELISA法测定血清和BALF中CC16水平,实时荧光定量RT-PCR检测肺组织CC16mRNA的表达,免疫组化法测定CC16在肺组织中的表达。
结果染毒1d组和30d组大鼠肺组织中CC16的含量明显高于对照组,染毒7d组大鼠肺组织中CC16的表达明显低于对照组和染毒1d组(P<0.01),染毒30d组大鼠肺组织中CC16的表达明显高于染毒1d组和7d组(P<0.01)；染毒7d组大鼠肺组织中CC16mRNA的表达高于对照组、染毒1d组和30d组(P<0.05)；染毒30d组大鼠血清中CC16的含量明显高于对照组、染毒1d组和7d组(P<0.05)；BALF中CC16的含量无明显变化。
结论
1.大气混合污染物早期炎症反应中(1 d),大鼠肺组织CC16明显高于对照组,而血清及BALF中CC16含量无明显变化,说明在早期炎症中有导致肺组织CC16增高趋势。
2.染毒7d时,大鼠肺组织当中的CC16明显低于对照组,说明肺组织细胞和毛细血管内皮细胞损伤严重。而CC16mRNA表达明显高于对照组,说明在损伤7d的过程当中CC16mRNA代偿性表达增强,CC16mRNA水平变化可作为早期肺损伤的敏感性生物学指标。
3.染毒30d时,血清中CC16含量明显升高,说明染毒后期肺泡组织自我修复过程及代偿性CC16合成增加导致血清中CC16含量明显增多。
4.肺组织中CC16含量降低可作为肺早期损伤的一个重要指标。血清CC16含量增高说明肺组织已进入一个修复期过程。BALF中CC16变化不明显说明BALF中CC16的含量不能作为判定肺组织损伤的一个参考指标。
With the rapid development of industrialization and urbanization, the usage of coal and oil fuel also increased strongly, then the urban air pollution problem become more prominent and constitute a badly threat to human health. The air pollutants such as particulate matter of atmosphere, SO2, NO2, CO and so on were closely related to the human health, among them, the PM2.5 was a kind of composite pollutant with complex components formed of large amount of different chemicals from man-made pollution and natural source, the toxicity of PM2.5 had prominent relationship with its chemical constituents,and heavy metal and charcoal were the main constituents of it. Air pollutants were closely related to human health, especially to the respiratory and circulatory system. Human clara cells were mainly distribute in epithelium of bronchiole terminates and respiratory bronchiole, CC16 was one of the 20 kinds of protein secreted by alveolar epithelium, and its main function was to prevent infringe of pollutants to alveolar epithelium, so the testing of CC16 was very valuable in evaluate injury of air-blood barrier or its integrity, it was very important in basic and clinical medical research.
Objective:
To explore the toxic effect mechanism of atmospheric pollutants (PM2.5, SO2, NO2, CO) to injury of lung tissue, this experiment mainly finded out the CC16 level in mouse blood serum and pulmonary tissue under the effect of mixed air pollutants via dustexposure and contamination method, in order to provide bases in preventing injury of air pollution to human body.
Methods:
78 Wistar rats were randomly divided into 6 guoups due to their weight,3 experiment groups (Id,7d,30d),3 control groups (1d,7d,30d),13 rats in each group.Rats were injected into non-open stance trachea with dust exposure method after anaesthetized with ether,1 ml saline suspl with 10 mg PM2.5 for experiment groups and 1 ml saline for control groups. Experiment groups were gived dynamic inhalation of air mixed with SO2, NO2, CO of 15,12, 400 mg/m3 for 4h per day, and lasted for Id,7d,30d in different experiment groups, and control groups were gived inhalation of normal air. ELISA method was utilized to determine the CC16 level in serum and BALF, real time RT-PCR method was used to detect the expression level of GC16 in lung tissue, immunohistochemistry method was used to determine the CC16 level in lung tissue.
Results:
1. CC16 expression in lung tissue of rats contaminated for Id and 30d were obviously higher than their control groups, AOD value had statistic significance (P<0.01). CC16 expression in lung tissue of rats contaminated for 7d was obviously lower than it's control group, AOD value had statistic significance(P<0.01). Comparison within experiment groups, CC16 expression in lung tissue of rats contaminated for 7d was obviously lower than that for Id, there was statistic significance (P< 0.01).CC16 expression in lung tissue of rats contaminated for 30d was obviously higher than that for Id, there was statistic significance (P < 0.01).CC16 expression in lung tissue of rats contaminated for 30d was obviously higher than that for 7d, there was statistic significance (P< 0.01).
2. CC16mRNA expression in lung tissue of rats contaminated for Id had no obviously change compared with it's control group, there was no statistic significance (P> 0.05). CC16mRNA expression in lung tissue of rats contaminated for 7d was higher than it's control group, there had statistic significance (P< 0.05).CC16mRNA expression in lung tissue of rats contaminated for 30d had no obviously change compared with it's control group, there was no statistic significance (P> 0.05). Comparison within experiment groups, CC16mRNA expression in lung tissue of rats contaminated for 1d and 30d were both lower than that for 7d, there had statistic significance (P< 0.05).
3. CC16 content in serum of rats contaminated for Id was a little lower than it's control group, but there was no statistic significance (P> 0.05).CC16 in serum of rats contaminated for 7d was a little lower than it's control group, but there was no statistic significance (P> 0.05). CC16 in serum of rats contaminated for 30d was obviously higher than it's control group and that had statistic significance (P< 0.05). Compared within experiment groups, CC16 in serum of rats contaminated for 7d was a little higher than that for 1d, but there was no statistic significance (P> 0.05), CC16 in serum of rats contaminated for 7d and 1d were obviously lower than that for 30d, that had statistic significance (P< 0.05).
4. CC16 content in BALF of rats contaminated for Id,7d and 30d had no obviously change compared with their control groups, there was no statistic significance (P> 0.05).CC16 in BALF of rats contaminated for Id,7d and 30d had no obviously change compared each other, there was no statistic significance (P> 0.05).
Conclusions:
1. In the early inflammatory reactions due to atmospheric pollutants (Id), the CC16 in lung tissue of rats were obviously higher than that in control groups but the CC16 in serum and BALF nearly had no change, then we can know that early inflammatory reactions may lead to increase of CC16 in lung tissue.
2. Endothelial Contaminated for 7d, the CC16 in rats'pulmonary tissue were obviously lower than that in control groups, that due to severe injury of lung tissue cells and endothelial cells of capillaries. And the CC16mRNA expressions were obviously higher than that in control groups expressed that in the 7d injury, the CC16mRNA expression compensatory increased,so the CC16mRNA level can be a kind of sensitive biologic index of early lung injury.
3. Contaminated for 30d, the CC16 in serum increased obviously, that means the self-repair of alveolus and the compensatory increase of CC16 compose which at last resulted in the CC 16 increase in serum.
4. The CC16 in lung tissue decrease was an important index of early lung injury and the increase of CC16 express that the pulmonary tissue had entered into a repairing process. Unchanged CC 16 in BALF means that the CC16 in BALF can not be an index in determinant of lung injury.
方法本研究在乙醚麻醉下,通过将实验组大鼠注入含有PM2.5的生理盐水混悬液和吸入含有SO2、NO2、CO的空气混合气来建立动物染尘染毒模型。应用ELISA法测定血清和BALF中CC16水平,实时荧光定量RT-PCR检测肺组织CC16mRNA的表达,免疫组化法测定CC16在肺组织中的表达。
结果染毒1d组和30d组大鼠肺组织中CC16的含量明显高于对照组,染毒7d组大鼠肺组织中CC16的表达明显低于对照组和染毒1d组(P<0.01),染毒30d组大鼠肺组织中CC16的表达明显高于染毒1d组和7d组(P<0.01)；染毒7d组大鼠肺组织中CC16mRNA的表达高于对照组、染毒1d组和30d组(P<0.05)；染毒30d组大鼠血清中CC16的含量明显高于对照组、染毒1d组和7d组(P<0.05)；BALF中CC16的含量无明显变化。
结论
1.大气混合污染物早期炎症反应中(1 d),大鼠肺组织CC16明显高于对照组,而血清及BALF中CC16含量无明显变化,说明在早期炎症中有导致肺组织CC16增高趋势。
2.染毒7d时,大鼠肺组织当中的CC16明显低于对照组,说明肺组织细胞和毛细血管内皮细胞损伤严重。而CC16mRNA表达明显高于对照组,说明在损伤7d的过程当中CC16mRNA代偿性表达增强,CC16mRNA水平变化可作为早期肺损伤的敏感性生物学指标。
3.染毒30d时,血清中CC16含量明显升高,说明染毒后期肺泡组织自我修复过程及代偿性CC16合成增加导致血清中CC16含量明显增多。
4.肺组织中CC16含量降低可作为肺早期损伤的一个重要指标。血清CC16含量增高说明肺组织已进入一个修复期过程。BALF中CC16变化不明显说明BALF中CC16的含量不能作为判定肺组织损伤的一个参考指标。
With the rapid development of industrialization and urbanization, the usage of coal and oil fuel also increased strongly, then the urban air pollution problem become more prominent and constitute a badly threat to human health. The air pollutants such as particulate matter of atmosphere, SO2, NO2, CO and so on were closely related to the human health, among them, the PM2.5 was a kind of composite pollutant with complex components formed of large amount of different chemicals from man-made pollution and natural source, the toxicity of PM2.5 had prominent relationship with its chemical constituents,and heavy metal and charcoal were the main constituents of it. Air pollutants were closely related to human health, especially to the respiratory and circulatory system. Human clara cells were mainly distribute in epithelium of bronchiole terminates and respiratory bronchiole, CC16 was one of the 20 kinds of protein secreted by alveolar epithelium, and its main function was to prevent infringe of pollutants to alveolar epithelium, so the testing of CC16 was very valuable in evaluate injury of air-blood barrier or its integrity, it was very important in basic and clinical medical research.
Objective:
To explore the toxic effect mechanism of atmospheric pollutants (PM2.5, SO2, NO2, CO) to injury of lung tissue, this experiment mainly finded out the CC16 level in mouse blood serum and pulmonary tissue under the effect of mixed air pollutants via dustexposure and contamination method, in order to provide bases in preventing injury of air pollution to human body.
Methods:
78 Wistar rats were randomly divided into 6 guoups due to their weight,3 experiment groups (Id,7d,30d),3 control groups (1d,7d,30d),13 rats in each group.Rats were injected into non-open stance trachea with dust exposure method after anaesthetized with ether,1 ml saline suspl with 10 mg PM2.5 for experiment groups and 1 ml saline for control groups. Experiment groups were gived dynamic inhalation of air mixed with SO2, NO2, CO of 15,12, 400 mg/m3 for 4h per day, and lasted for Id,7d,30d in different experiment groups, and control groups were gived inhalation of normal air. ELISA method was utilized to determine the CC16 level in serum and BALF, real time RT-PCR method was used to detect the expression level of GC16 in lung tissue, immunohistochemistry method was used to determine the CC16 level in lung tissue.
Results:
1. CC16 expression in lung tissue of rats contaminated for Id and 30d were obviously higher than their control groups, AOD value had statistic significance (P<0.01). CC16 expression in lung tissue of rats contaminated for 7d was obviously lower than it's control group, AOD value had statistic significance(P<0.01). Comparison within experiment groups, CC16 expression in lung tissue of rats contaminated for 7d was obviously lower than that for Id, there was statistic significance (P< 0.01).CC16 expression in lung tissue of rats contaminated for 30d was obviously higher than that for Id, there was statistic significance (P < 0.01).CC16 expression in lung tissue of rats contaminated for 30d was obviously higher than that for 7d, there was statistic significance (P< 0.01).
2. CC16mRNA expression in lung tissue of rats contaminated for Id had no obviously change compared with it's control group, there was no statistic significance (P> 0.05). CC16mRNA expression in lung tissue of rats contaminated for 7d was higher than it's control group, there had statistic significance (P< 0.05).CC16mRNA expression in lung tissue of rats contaminated for 30d had no obviously change compared with it's control group, there was no statistic significance (P> 0.05). Comparison within experiment groups, CC16mRNA expression in lung tissue of rats contaminated for 1d and 30d were both lower than that for 7d, there had statistic significance (P< 0.05).
3. CC16 content in serum of rats contaminated for Id was a little lower than it's control group, but there was no statistic significance (P> 0.05).CC16 in serum of rats contaminated for 7d was a little lower than it's control group, but there was no statistic significance (P> 0.05). CC16 in serum of rats contaminated for 30d was obviously higher than it's control group and that had statistic significance (P< 0.05). Compared within experiment groups, CC16 in serum of rats contaminated for 7d was a little higher than that for 1d, but there was no statistic significance (P> 0.05), CC16 in serum of rats contaminated for 7d and 1d were obviously lower than that for 30d, that had statistic significance (P< 0.05).
4. CC16 content in BALF of rats contaminated for Id,7d and 30d had no obviously change compared with their control groups, there was no statistic significance (P> 0.05).CC16 in BALF of rats contaminated for Id,7d and 30d had no obviously change compared each other, there was no statistic significance (P> 0.05).
Conclusions:
1. In the early inflammatory reactions due to atmospheric pollutants (Id), the CC16 in lung tissue of rats were obviously higher than that in control groups but the CC16 in serum and BALF nearly had no change, then we can know that early inflammatory reactions may lead to increase of CC16 in lung tissue.
2. Endothelial Contaminated for 7d, the CC16 in rats'pulmonary tissue were obviously lower than that in control groups, that due to severe injury of lung tissue cells and endothelial cells of capillaries. And the CC16mRNA expressions were obviously higher than that in control groups expressed that in the 7d injury, the CC16mRNA expression compensatory increased,so the CC16mRNA level can be a kind of sensitive biologic index of early lung injury.
3. Contaminated for 30d, the CC16 in serum increased obviously, that means the self-repair of alveolus and the compensatory increase of CC16 compose which at last resulted in the CC 16 increase in serum.
4. The CC16 in lung tissue decrease was an important index of early lung injury and the increase of CC16 express that the pulmonary tissue had entered into a repairing process. Unchanged CC 16 in BALF means that the CC16 in BALF can not be an index in determinant of lung injury.
引文
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