Study On Biomolecules For Early Coal Workers' Pneumoconiosis

Objective By investigating the general demographic information of Coal worker's pneumoconiosis and dust exposure and measuring the levels of serum cytokines of two groups of people,the risk factors of CWP and the changes of cytokine expression during the pathogenesis course of CWP were analyzed.A549 cells were exposed to coal dust and silica dust respectively.The cytotoxic effects and the expression of cytokines on A549 cells were analyzed.The differentially expressed cytokines were identified by statistical analysis,and the molecular mechanism of differentially expressed cytokines was preliminarily explored.Methods An epidemiological questionnaire was designed,and 69 CWP patients and 67 coal mine dust workers who came to the occupational disease prevention and control hospital in Ningxia for physical examination were investigated.Information from the survey includes surname and personal name,gender,date of birth,types of past work and start and end times of each job,daily working hours,diploma,smoking years and daily smoking amount,drinking time and monthly drinking amount,chronic respiratory symptoms,previous medical illness,current illness,whether contact dust,dust time and so on.Serum samples were collected from CWP and dust exposure groups.The serum expression levels of Tumor necrosis factor alpha(TNF-?),Interleukin 6(IL-6),Superoxide dismutase(SOD),malondialdehyde(MDA)and acetylcholinesterase(AChE)were determined.The risk factors of CWP were analyzed by binary logistic regression analysis.A549 lung adenocarcinoma cells were cultured under aseptic conditions.A549 cells were exposed to 0,50,100,200,400,600 ug/mL of coal dust and silica dust for 24,48 and 72 hours respectively.The effects of coal dust and silica dust on the A549 cell were judged by CCK8 assay determining the survival rate of A549 cells at different concentration of toxicants and at different exposure time.A549 cells were exposed to 0,50,100,200,400,600 ug/mL coal dust for 24 hours.The expression levels of TNF-?,IL-6,SOD,MDA,AChE,Butyrylcholinesterase(BChE)in A549 cells were determined.The levels of TNF-? and IL-6 in human serum and A549 cells were determined by enzyme-linked immunosorbent assay.The expressions of SOD and MDA in human serum and A549 cells were determined by colorimetry.The levels of AChE in human serum and A549 cells were determined by modified ELLMAN method.The levels of BChE in A549 cells were determined by modified ELLMAN method.A549 cells were exposed to 0,50,100,200 ug/mL coal dust.The expression levels of CHRNB2,CHRM5,CHRNA5,CHRNA7 and CHRNA9 were determined by RT-qPCR.Results The age,working years and dust exposure time of CWP group were higher than those of dust exposure group,while smoking index and drinking index were lower than those of dust exposure group.The results of Mann-Whitney U test showed that there were significant differences in these variables between the two groups(P < 0.05).Chi-square test showed that the two groups had different educational level,chronic respiratory symptoms and past medical history distribution(P < 0.05).On the whole,the educational level of dust exposure group was slightly higher than that of CWP group.The chronic respiratory symptoms of dust exposure group were lower than that of CWP group,and the detection rate of past medical history was higher than that of CWP group.With the progress of CWP lesions,the expression levels of IL-6,SOD and MDA in dust exposure group,CWP-1 phase and CWP-1 phase were significantly different(P < 0.05),while the expression levels of TNFalpha and AChE were not significantly different(P > 0.05).From dust exposure group to CWP ?stage and CWP ?stage,the expression levels of IL-6 and MDA increased,SOD and AChE decreased,and TNF-a increased first and then decreased slightly.Logistic regression analysis showed that the OR 95% CI value range of IL-6,MDA,advanced age was greater than 1,and that of AChE was less than 1.When coal dust acted on A549 cells,the cell survival rate decreased with the increase of toxicants.The variance analysis of factorial design showed that there were statistical differences between dose and time effects(P < 0.05),and there was no interaction between time and dose(P > 0.05).There was no significant difference in the time effect and dose of silica dust on A549 cells(P > 0.05),nor was there any interaction between time and effect(P > 0.05).The results of variance analysis showed that the changes of IL-6,MDA,TNF-?,SOD and BChE in A549 cells were not significant(P > 0.05),but AChE in A549 cells had significant difference(P = 0.044).There was no significant difference in CHRNB2,CHRM5,CHRNA5 and CHRNA9 among groups exposed to different doses of coal dust(P > 0.05),and there was significant difference in the expression of CHRNA7 among groups(P < 0.05).Conclusion The expression levels of IL-6 and MDA in human serum increased with the progression of CWP,while SOD and AChE decreased.Advanced age,high expression of IL-6 and MDA in human serum are risk factors for CWP,and high expression of AChE is a protective factor for CWP.Coal dust is cytotoxic to A549 cells.With the increase of coal dust dose,the survival rate of A549 cells decreases.Silicon dust has no obvious cytotoxicity to A549 cells.The changes of IL-6,MDA,TNF-alpha,SOD and BChE in A549 cells stimulated by coal dust for 24 hours were not significant among the exposed groups.The changes of AChE in the exposed groups were statistically significant,and the expression of CHRNA7 gene mRNA was significantly different among the exposed groups.