模拟月尘暴露致大鼠肺损伤作用研究
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摘要
目的研究模拟月尘(lunar dust simulant,LDS)致大鼠肺损伤的作用及其机制。方法采用CLDS-i模拟月尘,无菌生理盐水配制模拟月尘混悬液。Wistar大鼠随机分为LDS高、中、低剂量3个暴露组(气管内滴注,每周2次,持续3周),以及滴注生理盐水的对照组。在支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中进行白细胞分类计数和细胞因子检测,肺组织经HE染色和Masson染色后观察肺脏的病理变化,免疫组化测定α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)表达,同时测定氧化应激生物标志物含量。结果 LDS暴露组BALF中性粒细胞数量高于对照组(P<0.05)。LDS高剂量组肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)和白细胞介素-6(interleukin-6,IL-6)含量显著增加(P<0.01),肺部氧化-抗氧化失衡。病理组织染色及免疫组化结果显示,炎性病变程度及纤维化程度随LDS剂量增加而增强,LDS组大鼠肺组织中α-SMA表达水平显著升高(P<0.05,P<0.01)。结论炎症和氧化应激是LDS诱导大鼠呼吸系统损伤的重要机制。二者相互促进,可引起肺血管内皮细胞损伤和肺部炎症,导致肺间质纤维化。
Objective To study the respiratory system injury and its mechanism of lunar dust simulant(LDS)in rats.Methods CLDS-i was used to simulate lunar dust.Simulated lunar dust suspension was prepared in sterile saline.Wistar rats were randomly divided into high,medium and low dose groups,intratracheal instillation,twice a week for 3 weeks,and control group was given normal saline.White blood cell counts and inflammation indicators detection were conducted in the bronchoalveolar lavage fluid(BALF).The lung tissues were examined by HE and Masson staining for the pathological changes of the lungs.The expression ofα-smooth muscle actin(α-SMA)was also determined by immunohistochemistry.And oxidative stress biomarkers were determined.Results The number of BALF neutrophils in LDS exposed group was higher than that in control group(P<0.01).LDS significantly increased the levels of tumor necrosis factor alpha(TNF-α)and interleukin-6(IL-6)in high dose group(P<0.01).The extent of inflammatory lesions and the degree of fibrosis increased with LDS dose.The activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in lung tissue of LDS group decreased and the content of malondialdehyde(MDA)increased significantly(P<0.05,P<0.01).Conclusion Inflammation and oxidative stress are important mechanisms of LDS-induced respiratory system injury in rats.They promote each other leading to pulmonary vascular endothelial cell damage and lung inflammation,leading to interstitial fibrosis.
Objective To study the respiratory system injury and its mechanism of lunar dust simulant(LDS)in rats.Methods CLDS-i was used to simulate lunar dust.Simulated lunar dust suspension was prepared in sterile saline.Wistar rats were randomly divided into high,medium and low dose groups,intratracheal instillation,twice a week for 3 weeks,and control group was given normal saline.White blood cell counts and inflammation indicators detection were conducted in the bronchoalveolar lavage fluid(BALF).The lung tissues were examined by HE and Masson staining for the pathological changes of the lungs.The expression ofα-smooth muscle actin(α-SMA)was also determined by immunohistochemistry.And oxidative stress biomarkers were determined.Results The number of BALF neutrophils in LDS exposed group was higher than that in control group(P<0.01).LDS significantly increased the levels of tumor necrosis factor alpha(TNF-α)and interleukin-6(IL-6)in high dose group(P<0.01).The extent of inflammatory lesions and the degree of fibrosis increased with LDS dose.The activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in lung tissue of LDS group decreased and the content of malondialdehyde(MDA)increased significantly(P<0.05,P<0.01).Conclusion Inflammation and oxidative stress are important mechanisms of LDS-induced respiratory system injury in rats.They promote each other leading to pulmonary vascular endothelial cell damage and lung inflammation,leading to interstitial fibrosis.
引文
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[17] Chen C,Yang S,Zhang M,et al.Triptolide mitigates radiation-induced pulmonary fibrosis via inhibition of axis of alveolar macrophages-NOXes-ROS-myofibroblasts[J].Cancer Biology and Therapy,2016,17(4):381-389.
[18] Hector A,Griese M,Hartl D.Oxidative stress in cystic fibrosis lung disease:an early event,but worth targeting?[J].Eur Respir J,2014,44(1):17-19.
[2] Darquenne C,Borja MG,Oakes JM,et al.Increase in relative deposition of fine particles in the rat lung periphery in the absence of gravity[J].J Appl Physiol,2014,117(8):880-886.
[3] Lam CW,Scully RR,Zhang Y,et al.Toxicity of lunar dust assessed in inhalation-exposed rats[J].Inhalation Toxicology,2013,25(12):661-678.
[4] James JT,Lam CW,Santana PA,et al.Estimate of safe human exposure levels for lunar dust based on comparative benchmark dose modeling[J].Inhal Toxicol,2013,25(5):243-256.
[5] Sun Y,Liu JG,Zheng YC,et al.Research on rat’s pulmonary acute injury induced by lunar soil simulant[J].J Chin Med Assoc,2018,81(2):133-140.
[6]孙燕,郑洪楠,张贤瑶,等.模拟月尘致大鼠肺脏急性损伤病理学变化[J].航天医学与医学工程,2017,30(2):149-151.Sun Y,Zheng H,Zhang X,et al.Pathological Changes of Acute Pulmonary Injury in Rats Induced by Tracheal Perfusion of Lunar Soil Simulant[J].Space Medicine&Medical Engineering,2017,30(2):149-151.
[7] Zheng Y,Wang S,Ouyang Z,et al.CAS-1lunar soil simulant[J].Adv Space Res,2009,43(3):448-454.
[8] Tang H,Li X,Zhang S,et al.A lunar dust simulant:CLDS-i[J].Adv Space Res,2016,59(4):1156-1160.
[9] Nemmar A,Hoet PH,Dinsdale D,et al.Diesel exhaust particles in lung acutely enhance experimental peripheral thrombosis[J].Circulation,2003,107(8):1202-1208.
[10] Mikuz G,Gschwendtner A.Value of bronchoalvoeolar lavage in the diagnosis of lung disease[J].Verh Dtsch Ges Pathol,2000,84:129-135.
[11] Sun Y,Liu JG,Kong YD,et al.Effects of lunar soil simulant on systemic oxidative stress and immune response in acute rat lung injury[J].Int J Pharmacol,2018,14(6):766-772.
[12]张慧云,杨靖,龙星男,等.TNF刺激肥大细胞IL-6分泌的信号转导通路[J].细胞与分子免疫学杂志,2011,27(8):829-831.Zhang HY,Yang J,Long XN,et al.Signal transduction pathway for TNF-induced IL-6release from mast cells[J].Chin J Cell Mol Immunol,2011,27(8):829-831.
[13] Wang G,Zhao J,Jiang R,et al.Rat lung response to ozone and fine particulate matter(PM2.5)exposures[J].Environ Toxicol,2015,30(3):343-356.
[14] Guo C,Xia Y,Niu P,et al.Silica nanoparticles induce oxidative stress,inflammation,and endothelial dysfunction in vitro via activation of the MAPK/Nrf2pathway and nuclear factor-κB signaling[J].Int J Nanomedicine,2015,10:1463-1477.
[15] Pilling D,Vakil V,Cox N,et al.TNF-α-stimulated fibroblasts secrete lumican to promote fibrocyte differentiation[J].Proc Natl Acad Sci USA,2015,112(38):11929-11934.
[16] Kandhare AD,Bodhankar SL,Mohan V,et al.Effect of glycosides based standardized fenugreek seed extract in bleomycin-induced pulmonary fibrosis in rats:Decisive role of Bax,Nrf2,NF-κB,Muc5ac,TNF-αand IL-1β[J].Chem Biol Interact,2015,237(6):151-165.
[17] Chen C,Yang S,Zhang M,et al.Triptolide mitigates radiation-induced pulmonary fibrosis via inhibition of axis of alveolar macrophages-NOXes-ROS-myofibroblasts[J].Cancer Biology and Therapy,2016,17(4):381-389.
[18] Hector A,Griese M,Hartl D.Oxidative stress in cystic fibrosis lung disease:an early event,but worth targeting?[J].Eur Respir J,2014,44(1):17-19.
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