The Study Of Proliferation, CTGF Expression And Biomechanical Properties Of Lung Fibroblasts After Treated By Paraquat

Paraquat is ono kind of phytocide 2, 2'-bipyfidine. Oral administration is the most common way to be poisoned, and the mortality is high. As so far the mechanism of poison is not clear. It was believed that a lot of oxygen radical caused cell membrane lipid peroxidize, which lead to multiorgan disturbed. The most common and scvere injury of lungs is pneumonedema and pulmonary fibrosis. Also there is no any antidote to paraquat poison. China is a big country which produce and use the agricultural chemicals. We have made a great progress in the reasonable remedy of the organophosphorus pesticide, but right now we also don't have any effect method to the parquet poisoning, it is emphasis of our experimentations and clinical task. Our experimentation is doing research in the proliferation of human embryonic lung fibroblast,viscoelasticity and the express of CTGF by Mechanics-biology method inorder to introduce the mechanism of the pneumonocyte impairment by parquet intoxation. We also want to find a new point for the effective therapeutic methords.Methods:The human embryonic lung fibroblast MRC-5 we use is bought from the Chinese Academy of Medical Sciences'cell repository. It is cultivanted in the MEM which includes 10% calf serum and made the serial subcultivation in the closed-circuit milclose whose temperature and humidity is constant, and the content of CO2 is 5%, the temperature is 37℃. We prepare different density of parquat MEM -50mg/L,100mg/L,200mg/L for use.then collect MRC-5, inoculate it in the six bore cultivation board and put the board in the closed-circuit milclose, 12h ago, we have to change the MEM which does have any parquet. (1) Use the MRC-5 which is disposed by the PQ, and make it being cultivated in the MEM without PQ for 24h. research the influence to the MRC-5 proliferation by PQ.(2) Use the MRC-5 which is disposed by the PQ, and make it being cultivated in the MEM without PQ, then collect the supernate until 12h,24h,36h,48h,60h,72h, and put them in refrigeration about -70℃after centrifugation to detect the express of the CTGF by the ELISA methord.(3) Use the MRC-5 which is disposed by the PQ, and make it being cultivated in the MEM without PQ for 24h. collect MRC-5 cells, make the cell suspension, then detect the viscoelasticity of the the human embryonic lung fibroblast MRC-5. Research the influence to the MRC-5 biomechanics by PQ.Result:The proliferation of the normal MRC-5 is much high and the proliferation of the MRC-5 which is disposed by the PQ is much low. The lower the PQ density is more. (2) The normal MRC-5 could secrete a little CTGF, the density of CTGF has been increasing with time in the first 48h (P<0.05). but they don't have any difference at 60h,72h and 48h (P>0.05); Use the MRC-5 which is disposed by the 50mg/L PQ, it has secreted a little CTGF in the first 36h, and it has no difference, but it has increased very much until 48h (P<0.05),the density of CTGF has been increasing with time from 48h to 72h. Use the MRC-5 which is disposed by the 100mg/L PQ, it has secreted a little CTGF in the first 36h, and it also has no difference, but it has increased very much until 48h (P<0.05), the density of CTGF has been increasing with time from 48h to 72h, Use the MRC-5 which is disposed by the 200mg/L PQ, the result is also just the the 100mg/L PQ. (3) The viscoelasticity parameters of the MRC-5 which is disposed by PQ are obviously justo minor, also we can say that,the biomechanics of the MRC-5 disposed by PQ step down, the cells are become"soft". And the viscoelasticities of MRC-5 disposed by 100mg/L PQ and 200mg/L PQ are smaller than the viscoelasticity of MRC-5 disposed by 50mg/L PQ (P<0.05), but they don't have any difference between 100mg/L and 200mg/L (P>0.05).Conclusion:(1) The result we reach is that the proliferation of the normal MRC-5 is much high and the proliferation of the MRC-5 which is disposed by the PQ is much low. The lower the PQ density is more. (2) The CTGF secreted by MRC-5 which is disposed by the PQ after 72 hours is obviously justo minor. It hints that the activated fibroblasts could lead cell multiplication and ECM production by the overdose CTGF secretion. It is very important in the pulmonary fibrosis. (3) The viscoelasticity parameters of the MRC-5 which is disposed by PQ are obviously justo minor,the biomechanics of the MRC-5 disposed by PQ step down. The resut hints that in the toxicosis of PQ, the cytoskeleton has been destroyed. (4) The toxicosis of PQ could change the cytomechanics conducting system, it hints that we could treat it by contralling the the overexpressed CTGF and repairing the cytomechanics conducting system.