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大气混合污染物对大鼠呼吸道微生态影响的研究
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摘要
目的:大气污染是随着科学文明的发展和工业化进程的加快而产生的环境问题。已有足够证据表明,大气混合污染物对呼吸道微生态发生改变,并造成呼吸道上皮损伤。本研究目的在于进一步探讨大气混合污染物细颗粒物对呼吸道微生态的影响及炎性损伤的形态学变化。方法:采集细颗粒物(PM 2.5)模拟现实大气污染,实验使用78只Wistar大鼠,雌雄各半,分为6个组:3个实验组(1d、7d、30d),3个对照组(1d、7d、30d)。在染尘染毒前用咽试子于大鼠口咽部取材。大鼠在乙醚麻醉下进行非暴露式气管一次注入染尘,实验组大鼠分别注入1ml含10mgPM2.5的生理盐水混悬液,对照组大鼠注入1ml生理盐水。第二天,实验组进行动态吸入含有SO2、NO2、CO的空气混合气,对照组吸入正常空气。研究大鼠口咽部菌群经过染尘染毒前后的变化,运用透射电镜观察染尘染毒前后大鼠气管和肺组织的超微结构。
     结果:染尘染毒后多数菌属检出率明显增加(p<0.05),需氧菌、厌氧菌细菌总量有所增加,菌群密度也均有所增加,且变化具有显著性(P<0.01)。部分菌属在染尘染毒后出现。染尘染毒后1d,需氧菌菌群密度显著增加,明显高于对照组(P<0.05),7d时明显减少,30d时又有明显增加(P<0.05),随染尘染毒时间的延长有明显波动的趋势,表现出代偿状态。细菌总量的不同染尘染毒时间组比较亦表现出同样变化趋势,且具有显著性(P<0.05),但厌氧菌无显著性差异。染尘染毒使大鼠的气管及肺泡上皮有炎性损伤,随着染尘染毒时间的延长逐渐加重。
     结论:大气污染物细颗粒物可导致大鼠口咽部细菌总量及菌群密度增加,使呼吸道微生态发生改变,并在此基础上造成肺组织及气管上皮损伤,对机体产生危害,形态学变化与吸入污染物的时间长短有一致性。
Air pollution is the environmental problem with the development of science and civilization as well as the speeding up of industrialization, generated because of the dense urban population, and rapid increase of fossil energy and fuel combustion. The sources of air pollutants in the city are similar, mainly composed of particle pollutants and gaseous pollutants. According to the aerodynamics diameter, particulates may be divided into: total suspended particulates and inhalable particulates, of which the latter may be divided into coarse particulates (PM2.5~10) and fine particulates (PM2.5). PM2.5 is mainly originated from combustion of fossil fuel. With small grain diameter, large quantity and surface area, slow settlement speed, far dissemination distance, PM2.5 may provide carriers for chemicals, bacteria and viruses. PM settling in human body is primarily PM2.5. Due to different regions and sources of pollution, chemical compositions of PM2.5 are greatly different. Major components include sulfate, nitrate, ammonium, carbon particles, metal particles, minerals, and organic compounds adsorbed by them. Breathing is one of the main routes by which suspended particulates and other harmful substances in the air enter into human body, and respiratory system is the direct target organ that air pollutants exert effect on. The number of residents who suffer from pharyngeal irritation and much sputum increases in areas with heavy air pollution compared with areas having light pollution, and the body's immune function may be damaged by repetitive long-term irritation of environmental pollutants with low concentration.
     At present, domestic and foreign research on air pollution is only restricted in aspects such as health investigation, dangerous factor analysis and pollution governance. In the earlier research, we have simulated the atmospheric pollutant mixture, established the animal model of air pollution, and discussed the effect of air pollution on the micro ecology of respiratory tract: air pollution (PM10) may cause unbalance of micro ecology in organisms, exert toxic and harmful effect on the normal bacteria colony in the oropharynx of upper respiratory tract in rats, resulting in the change of micro environment. The content of normal bacteria colony reduced, the pathogenic bacteria and the conditioned pathogen colonized to facilitate the occurrence of diseases in the respiratory tract. Air pollution may lead to increased bacteria density in the oropharynx of rats, increased pathogen number and the change of bacteria number, and the change of micro ecology in respiratory tract may be taken as the sensitive indicator of harm by air pollution to organisms. In the further research, we mainly discussed the effect of fine particulates of atmospheric pollutant mixture on the micro ecology of respiratory tract. we mainly gathered smaller inspirable particles (PM 2.5), and used 78 Wistar rats in the experiment, divided into 6 groups: 3 experimental groups (1d, 7d, 30d) and 3 control groups (1d, 7d, 30d). Polluted dusts were poured into the trachea of rats one time in the non-exposed manner under the ether anesthesia. Materials were taken from the oropharynx of rats with pharynx swab before it was contaminated by polluted dusts. In experimental rats, 1ml 10mg/PM2.5 normal saline was injected, and in control rats 1ml normal saline was injected. Air mixture with SO2 and NO2 and CO was inhaled dynamically by the experimental group, and normal air inhaled by the control group. The similarities and differences before and after the oropharynx of rats was contaminated by polluted dusts were studied, the ultra-structure of the trachea and pulmonary tissue of rats before and after contamination by polluted dusts was observed with the transmission electron microscope
     After contamination by polluted dusts, detected ratios of most bacteria increased significantly (p< 0.05), the total quantity of aerobes and anaerobes increased, and colony density also increased, and the change is significant (P< 0.01). Partial bacteria appeared after contamination. The colony density of aerobes increased remarkably 1 day after contamination by polluted dusts, significantly higher than the control group (P< 0.05), reduced obviously 7 days after contamination, and obviously increased 30 days after contamination (P< 0.05). With the prolonging of contamination time, it showed compensation state with obvious trend of fluctuation. The comparison between groups of different contamination time also showed the same change trend in terms of total quantity of bacteria, with significance (P< 0.05), but there was no significant difference in anaerobe. Contamination by polluted dusts caused inflammatory damage to the trachea and pulmonary alveolus of rats, which aggravated with the prolonging of contamination.
     The conclusion may be drawn that fine particulates of air pollutants may cause increased pathogenic bacteria in the oropharynx of rats, simultaneously the change of micro ecology results in the damage of pulmonary tissue and tracheal epidermis on the basis of pollution of fine particulates, therefore, fine particulates may change the micro ecology of respiratory tract and pose risk to organisms.
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