The Experimental Study Of Treatment For Acute Lung Injury Mediated By BMSCs

ObjectiveTo investigate the feasibility that lipopolysaccharide(LPS)make the acute lung injury animal model of rats.To investigate the isolation,culture amplification and identification of BMSCs.To analyze the distribution of BMSCs and restoration for the injured lung.To investigate the possibility that BMSCs treat severe diseases of the lung and provide the reference of treatment for severe diseases of the lung.MethodsThe femur and tibia of Wistar rats of six weeks(150g)were Dissected and the bone marrow was lavaged into the centrifugation tube.The bone marrow was made into suspension which was centrifuged repeatedly.Then the suspension was mixed with Percoll solution and centrifuged.The BMSCs were collected and inoculated. The culture medium was first replaced after 24 hours and then was replaced every 3 days.The BMSCs were digested.We used the second passage cells to detect cell surface markers by the immune fluorescence assay.Twenty rats were divided into two gtoups:A group injected with only LPS and B group injected with both LPS and BMSCs marked with BrdU Bromodeoxyuridine.The respiratory condition, respiratory frequency,activity,eating and secretion in mouth and nose were observed. After 5 days,the rats were killed.The lungs of the rats were dissected and examined pathologically.The results were analyzed and compared.Results1,The primary cultured BMSCs get 80-90%confluence within 9-10days.The shape of the offspring cells had homogeneity. 2,The cell surface markers CD29 and CD44 were positive,whereas CD34 and CD45 were negative through the immune fluorescence assay.3,The respiratory conditions and activities of the control rats were good.Some rats of both groups had dyspnea,hyperpnea,hypoactivity,fasting and secretion in mouth and nose.However,the number of suffered rats in A group are more.The difference of two groups were significant statistically(P＜0.05).4,The lungs of the control rats were normal.The lungs of both groups showed congestion and edema.However,the lungs of A group were severer,in which 3 lungs had visible consolidation macro pathologically.The pathological slices of the rats injected with LPS(A group)showed edema,congestion and consolidation of lungs. The pathological slices of the rats injected with LPS and BMSCs(B group)showed mild edema and congestion of lungs.Also,they did not show consolidation of lungs.5,The pathological slices of the rats injected with only LPS(A group)were negative and positive with both LPS and BMSCs(B group)marked with BrdU Bromodeoxyuridine by the immunohistological examination.Conclusion1,The lung injury of the rats injected with both LPS and BMSCs is milder than that with LPS only.2,The BMSCs injected into the rats with acute lung injury can distribute in both the lungs of the rats.