| Objective: Paraquat is a quarternary nitrogen herbicide used for broadleaf weed control and is widely used in more than 140 countries and regions including China. Suicide and accidents caused by PQ poisoning showed an increasing trend in recent years. The damage on the body of PQ involves a variety of pathological mechanisms, and currently the effect of paraquat-induced peroxidation damage has been caused enough attention. Because there is no specific detoxification drug on paraquat and the treatment strategies are being explored, antioxidants are considered to be new drugs with good prospects. 5-amino salicylic acid is a classic drug to treat inflammatory bowel disease. It has good effect of scavenging oxygen free radicals, reducing Lipoxygenase activity, inhibiting prostaglandin synthesis, and so on.This study is to extend our previous study through the measurement of sod inhibition rate and the content of MDA and GSH to explore the influence of paraquat concentration on WI-38VA13 cells, and further discussion of the possible protection mechanisms of 5- ASA on acute lung injury induced by PQ.Method: The WI-38VA13 cells were routinely cultured in DMEM/F12 supplemented with 100U/ml penicillin, 100 U/ml strep-tomycin, and 10% fetal bovine serum(FBS), under 5% CO2 /95% air. The cells were treated with solvent alone or with various concentrations of PQ for 24 h,then were measured survival rate by MTT and selected appropriate PQ and suitable 5-ASA concentrations. WI-38VA13 cell are handled as follows: WI-38VA13 cells were divided into PQ 0 μ mol/l, PQ 100 μ mol/l group, PQ 200 μ mol/l group, PQ 400 μ mol/l group,PQ 600μ mol/l group and PQ 800 μ mol/l group, after were routinely cultured at 5% CO2 /95% air for 24 hours. After WI-38VA13 cells were exposed in PQ 200 μ mol/l. Then the cells were divided into 5-ASA 0m M the intervention group, 5-ASA 5m M intervention group, 5-ASA 10 m M intervention group intervention after treatment at 5% CO2 /95% air for 24 hours. The total protein from each group were collected. And each group were measured inhibition of SOD and contents of MDA, GSH. For each protocol, at least three independent experiments were performed. Results were expressed as the mean ± standard error of the mean(SEM). Statistical calculations were performed with the one way analysis of variance(ANOVA) using SPSS19.0 software. Differences in measured variables between experimental and control groups were assessed by Regression and paired-samples t-test. P<0.05 was considered statistically significant.Results: 1 Effect of the PQ concentration on survival rate of lung fibroblast cellsThe survival rate of each concentration group declined. 200 μ mol/l PQ to 1000 μ mol/l PQ groups compared with the control group, WI-38VA13 cell survival rate was significantly decreased(P<0.05). 2 Effect of the 5-ASA concentration on survival rate of lung fibroblast cellsWI-38VA13 cells were incubated in the different concentration of 5-ASA for 24 h. We found that the survival rate of 20 Mm 5-ASA and 30 m M 5-ASA groups significantly decreased compared with the control group(P<0.05). 3 Effects of the PQ concentration on the oxidative damage of lung fibroblast cells 3.1 Effect of the PQ concentration on SOD inhibitory rate of the lung fibroblast cellsConcentration inhibition rate of SOD was all increased. 200 μmol/L group, PQ 400 μmol/L group, PQ 600 μmol/L group, and PQ 800 μmol/L group compared with solvent control groups, SOD inhibition rate significantly increased(P <0.05). 3.2 The level of MDA of the lung fibroblast cells in different paraquat concentrationThe concentration MDA content had increased. MDA content of PQ 200 μmol/L group, PQ 400 μmol/L group, PQ 600 μmol/L group, and PQ 800 μmol/L group were significantly higher than the control groups(P<0.05). 3.3 The level of GSH of the lung fibroblast cells in different paraquat concentrationGSH concentration of each groups were reduced in various degrees. PQ 200 μmol/L group, PQ 400 μmol/L group, PQ 600 μmol/L group and PQ 800 μmol/L group compared with control group, levels of GSH were significantly decreased(P<0.05). 4 Interventional effect of 5-ASA with different concentrations to PQ-exposed lung fibroblast cells 4.1 Effect of different concentrations of 5-ASA on SOD inhibitory rate of PQ-exposed lung fibroblast cellsThe SOD inhibition rates of 5-ASA different concentrations all reduced in various degrees. 5-ASA 5 m M intervention group and 5-ASA 10 m M intervention group been compared with 5-ASA 0 m M intervention group, SOD inhibition rate were significantly lower than 5-ASA 0 m M the intervention group(P<0.05). The SOD inhibitory rate of 5-ASA 10 m M intervention group was more pronounced(P<0.05). 4.2 The level of MDA of PQ-exposed lung fibroblast cells in different concentrations of 5-ASA5-ASA 5 m M intervention group and 5-ASA 10 m M intervention group been compared with 5-ASA 0 m M intervention group, MDA significantly lower levels of 5-ASA 0m M intervention group(P<0.05). 5-ASA 10 m M MDA had lower levels of intervention group(P<0.05). 4.3 The level of GSH of PQ-exposed lung fibroblast cells in different concentrations of 5-ASA5-ASA 5m M and 5-ASA 0m M, 5-ASA 10 m M intervention group intervention group intervention group compared,GSH content ware significantly higher than that 5-ASA 0m M intervention group(P<0.05). 5-ASA 10 Mm intervention group GSH had lower levels of intervention group(P<0.05).Conclusions:1 Paraquat can cause lung fibroblast cells peroxidation injury, and with the higher paraquat concentration, the more serious injury is formed.2 Proxidation in acute lung injury is an important mechanism induced by paraquat. When PQ exposed on lung fibroblast cells, SOD inhibition rate increased, lipid peroxidation products MDA increased and antioxidant GSH reduced.3 It is a positive correlation between the concentration of paraquat to lung fibroblasts and the extent of oxidative damage. With the higher the concentration of paraquat, the degree of oxidative damage to lung fibroblasts increased.4 The proper concentration of 5-ASA can resist oxidation, reduce lipid peroxidation, and has protective effect to paraquat-induced lung injury. |
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