| Objective:Based on the previous study,in order to explore the correlation between fine particles and harmful substances released from C1 coal combustion in Xuanwei area,Yunnan Province,and the incidence of injury of lung in Xuanwei,a female F344 rat lung cancer model was established.The genes related to lung injury in F344 rats were screened by gene transcription group sequencing,which laid a foundation for studying the mechanism of lung cancer caused by the burning and releasing of fine particles and harmful substances from C1 coal in Xuanwei area.Methods:1.To establish the injury of lung model of female F344 rats:two-week-old female F344 rats were selected as experimental objects to induce C1 coal in Xuanwei area,Yunnan Province,and the experimental rats were divided into experimental group and control group.The experimental group F344 rats were exposed to C1 coal combustion products by a self-made laboratory device simulating "open fire" in Xuanwei area.The exposure time was 24 weeks after inhalation of C1 bituminous coal combustion products for 20 hours a week.The control group only inhaled fresh air.2.Pathological sections were made:at 4 weeks,8 weeks,12 weeks,16 weeks and 24 weeks,6 rats in the experimental group and 2 rats in the control group were randomly selected.After anesthesia,the rats were dissected and left with right lung,trachea,thymus,heart and liver.The spleens were embedded in paraffin and stained with HE.3.Sequencing in the transcriptional group:at 4 weeks,8 weeks,12 weeks,16 weeks and 24 weeks,3 rats were randomly selected from the left lung tissue of the experimental group and 1 rat of the control group was sent to Huada gene to sequence the gene in the transcriptional group.5.Determination of serum cytokines:rat INF-γ,IL-2,IL-10,IL-17 ELISA kit were used for elisa test.Results:1 Changes of general situation of female F344 rats induced by C1 coal products of Xuanwei area,Yunnan.1.1 General conditions of rats.During the experiment,no death occurred in the experimental group F344 rats and the control group F344 rats.With the prolongation of the exposure time of coal combustion,the activity,mental state,food intake and body weight of the F344 rats in the experimental group were decreased compared with those in the control group,there is no obvious change in the quantity of drinking water.The body hair was obviously yellowed,pale and lustrous compared with the control group.The control group F344 rats had normal activity,good mental state,normal intake of water,and normal weight gain,no body hair yellowed or dimmed.1.2 The gross tissue of female F344 rats induced by C1 coal combustion products in Xuanwei area of Yunnan Province.No obvious pulmonary nodules were found in the lung tissues of F344 rats at 4 weeks,but compared with the control group,the lung tissue was dark.At 8 weeks,12 weeks,16 weeks and 24 weeks,the lung tissue of the experimental group was darker than that of the control group,and the lung tissue of the experimental group was darker than that of the control group,appear dark red.In 60%of the experimental rats,there were dark plaques visible to the naked eye and a large number of small nodules could be touched in the gross tissues of the lungs of the rats.At 24 weeks,the lung tissue of one of the rats in the experimental group could touch a larger mass.In the control group,there was no significant abnormality in the lung tissue of the F344 rats.The other tissues and organs(trachea,heart,thymus,liver and spleen)were not abnormal in the experimental group and the control group.1.3 The microscopic tissue of female F344 rats induced by C1 coal combustion products in Xuanwei area,Yunnan ProvinceIn the experimental group,the wall of bronchus and bronchioles was slightly thickened at 4 weeks in F344 rats,but the shape and structure of the wall are still relatively intact.Lymphocytes,macrophages and other inflammatory cells could be seen around the wall of bronchus.The alveolar cavity was slightly enlarged,and there was a tendency of fusion.At 8 weeks,12 weeks,20 weeks,24 weeks,a large amount of carbon membrane deposition,bronchial wall thickening and mucosal damage were observed in the lung tissue of the experimental group rats,cilium lodging and shedding of mucosal epithelium,a large number of lymphocytes and macrophages are accumulated around the wall of the tube.Alveolar wall thinning,part of alveolar wall can be seen fracture fusion into alveolar bullae,partial infiltration of inflammatory cells around alveolar wall;partial carbon deposition and partial aggregation of heterocyt cells were observed near the pleura.At 24 weeks,an experimental group had a rat with inflammatory pseudotumor under microscope.In the control group,there was no lung tissue damage,no infiltration of lymphocytes and macrophages,and no deposition of carbon membrane near the pleural membrane in the control group.2 Sequencing results of gene transcripts of F344 rats induced by burning products of Xuan Wei C1 coal in Yunnan province2.1 The quality of sequencing data of the F344 rats gene transcriptional groupThe gene transcript group of 24 F344 rats was successfully constructed by RNA-seq technique.The quality statistics of filtered reads were all qualified.2.2 Analysis of gene information of F344 rat gene transcriptional set sequencingBased on the gene expression of each sample transcript group,we compared the gene expression between the experimental group.and the control group.Through the analysis,we found that there are different genes up-regulated and down-regulated in each pair of samples.The results of these genes are consistent with those related to chronic lung injury.Go and KEGG analysis of the differentially expressed genes between the experimental group and the control group showed that there was significant enrichment of GO term and KEGG pathway in different degrees among the samples.It mainly includes immune response,inflammatory response,apoptosis,signal transduction and so on.3.Changes of cytokines in serum of F344 rats induced by burning products of Xuanwei C1 coal4 weeks,8 weeks,16 weeks,20 weeks and 24 weeks,the concentration of INF-γ、IL-2、IL-10、IL-17in the experimental group was higher than that in the control group,and the difference was statistically significant.At 12 weeks,the concentration of INF-γ、IL-2、IL-10、IL-17 in serum of F344 rats was higher than that of control group,but there was no significant difference.Conclusion:1.The products released from C1 coal combustion in Xuanwei area of Yunnan Province resulted in the changes of lung tissue and cell structure in female F344 rats,resulting in heteromorphism,suggesting that the pathogenesis of lung cancer may be related to indoor exposure to bituminous coal.2.The differential gene Ahrr,Armcxl,Nrldl,and immune system related differential gene CD52,Wfdc21,related to lung cancer were successfully screened by ma-seq to speculate the lung damage caused by C1 bituminous coal combustion products in Xuanwei area,Yunnan Province.It may be related to the imbalance of immunomodulatory and tumor-promoting gene expression in rats.3.GO,KEGG analysis of the differentially expressed genes and cytokine analysis of rat serum showed that the long-term inhalation of C1 bituminous coal-released products may be associated with the imbalance of immunomodulation in the lungs of rats. |
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