HO-1 Modified Mesenchymal Stem Cells For Rat Heart-free Donor Lung Ischemia-Reperfusion Injury

Purpose To observe the effect of HO-1 gene-modified BMSC on pulmonary ischemia/reperfusion injury in rats without heartbeat and explore the mechanisms.Method First,an animal model of NHBD lung transplantation IRI was constructed in SD rats to simulate the case of clinical NHBD lung transplantation IRI.HO-1-BMSC was constructed by modifying BMSC with HO-1 as target gene and lentivirus as vector.Then HO-1-BMSC was injected into the receptor by pulmonary fine needle puncture.The operation control group(I),simple BMSC group(II),lentivirus empty vector BMSC group(III),HO-1 gene transfection group(IV)were established.In each group,n = 15,no heartbeat donor lung was retrograde perfused through pulmonary vein with LPDG at 4? and then stored at 4?-LPDG solution for 1 hour before implanted into recipient rat.After successful transplantation,group II,group III and group IV were injected with RFP-labeled BMSC,lentiviral empty vector-BMSC and HO-1-BMSC through pulmonary artery fine needle respectively.After 4 hours of reperfusion,abdominal aortic blood was taken for blood gas analysis when the right pulmonary porta was blocked.The changes of TNF-a and IL-10 in plasma of recipient rats were detected by ELISA.The W/D ratio,SOD activity,MDA content and MPO activity of lung tissue were measured.Histopathological changes of grafts in each group observed by light microscopy.RT-PCR and Western-blot were used to detect the expression level of HO-1 gene in each group.Immunofluorescence was used to detect BMSC colonization in the grafts.Results Group IV > Group II and Group III > Group I: Pa CO2 decreased,W/D ratio decreased,Pa O2 increased(P<0.05);MDA content and MPO activity decreased,SOD activity increased,(P<0.05);TNF-? expression decreased,IL-10 expression increased(P<0.05).Light microscopy showed that neutrophils infiltrated pulmonary interstitium and inflammatory exudation was seen in alveolar cavity.Inflammation in group IV was the lightest in each group.The highest expression of HO-1 was found in group IV.Under electron fluorescence microscopy,BMSC colonization was observed mainly on alveolar wall.Conclusion HO-1 gene modified BMSC has synergistic anti-injury effect in the treatment of IRI of non-heart-beating donor lung transplantation,which provides a new idea for clinical treatment of NHBD lung transplantation and even other transplantation.