| Objective By investigating the expression of Toll-like receptor(TLR4) in swine lung ischemia reperfusion injury(IRI) after lung transfertation,to explore the raletion between TLR4 and IRI, and to compare protective effect of "ex vivo oxygenated blood by continuous perfusion" and "classic static preservation in 4 ? temperature and preservative solution".Methods 12 couples of healthy Guangxi Bama miniature pigs were divided into two group randomly and equally. The donor lungs of test group(ex vivo oxygenated blood continuous perfusion for 8h) and control group (classic static preservation in 4? temperature for 8h) were both transplanted and reperfused. 1. Recorded the femoral artery PO2/FiO2 and the lower left pulmo-nary vein blood PO2/FiO2 (LIPV-PO2/FiO2) of donor pigs at 0.5 h post-transplantation and every two hours post-transplantation. 2.At the time point of before storaged the donor lung and after saved for 8 hours and 0 h, 3 h, 6 h, 12 h,24 h post-transplantation, (1)measured resistance of lung ventilation (LR) of left receptor lung, (2) collected the left donor lung tissue samples to measure lung water content to evaluate the lung injury, and observed the submicrostructure and the morphologic change of the lung apoptotic cell to evaluate the lung tissue injury, (3) used western blot and real-time PCR to detected quantitatively the changes of TLR4 and TLR4 mRNA expression in donor lung tissue.Results (1) The femoral artery PO2/FiO2 and LIPV-PO2/FiO2. The femoral artery PO2/FiO2 and LIPV-PO2/FiO2 decreased gradually occurred within 12-hours post-transplantation in both groups. By the first 24 hours post-transplantation, the control group decreased gradually and the text group increased significantly (P <0.05). The test group showed significantly higher levels of femoral artery PO2/FiO2 and LIPV-PO2/FiO2 at each time point post-transplantation. (2) LR and lung water content. LR and lung water content increased gradually occurred within 12-hours post-transplantation in both groups.By 24h, the control group increased gradually and the text group decreasedsignificantly. (3) Submicrostructure and the morphologic change of the lungapoptotic cell. The donor lung tissues began to apoptotic after saved 8h by transmission electron microscopy. We could observed all periods of form of cell apoptosis in both groups at 3h and 6h post-transplantation. At 12h post-transplantation, control group was discovered early apoptosis while text group had none. At 24h post-transplantation, both groups based form on late apoptosis.(4) The expression of TLR4 and TLR4 mRNA. Compared with before storaged the donor lung, TLR4 and TLR4 mRNA increased obviously and the test group showed higher level (P <0.05). Compared with after saved for 8 hours, TLR4 and TLR4 mRNA increased significantly at 3h post-transplantation (P <0.05).At the 6h post-transplantation, the expression of TLR4 and TLR4 mRNA reached the peak in the test group, then it was down-regulated at 12h and 24h post-transplantation. However, the expression of TLR4 and TLR4 mRNA had no difference between 6h and 12h post-transplantation in the control group (P>0.05), TLR4 and TLR4 mRNA at 24h post-transplantation was significantly lower than that at 24h post-transplantation (P <0.05). Furthermore, the test group showed significantly lower levels of the expression of TLR4 and TLR4 mRNA at each time point post-transplantation.Conclusion (1) Both groups could induce damage of lung injury after saved the donor lung for 8 hours. Damage to lung function injury persisted within 24 hours in "classic static preservation in 4 ? temperature", while it was improved gradually after 12h post-transplantation in "ex vivo oxygenated blood by continuous perfusion in 36?". (2) Ex vivo oxygenated blood continuous perfusion always showed lower expression of TLR4 than "classic static preservation in 4 ? temperature", which had same trend with lung injury.Therefore, Ex vivo oxygenated blood continuous perfusion may be a better lung preservation techniques. |
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