The Primary Research In The Therapy Effect Of Embryonic Stem Cells In Injury Of Mouse Lung Induced By Bleomycin

Objective The aim of this study was to observe and compare the therapy effect of different kinds of embryonic stem cells (ESCs) including C57BL/6J-ESC, S8-ESC and human-ESC on pulmonary injury of mice exposed to bleomycin. The cells with best effect was hoped to be picked out and transplanted twice to the mice exposed to bleomycin for further study.Methods I. Looking for the dose of bleomycin to induce lung injury of mice. 1. Forty C57BL/6J female mice were divided randomly into four groups including blank control group, model group with 6mg/kg bleomycin, 8mg/kg bleomycin and 10mg/kg bleomycin. 2. The mice of the control group were administrated 0.9% sodium chloride solution and the mice in other three groups were administrated 6mg/kg, 8mg/kg and 10mg/kg bleomycin transtracheally. 3. The life-span and body weight changes and the pathologic changes of the lung were observed. II. The primary research of the therapeutic effect of embryonic stem cells on the injury of mouse lung induced by bleomycin. 1. Fifty C57BL/6J female mice were divided randomly into five groups, they were blank control group, bleomycin model group, bleomycin model with C57BL/6J-ESC, bleomycin model with S8-ESC and bleomycin model with human-ESC. 2. The mice of the control group were administrated 0.9% sodium chloride solution and the mice in other four groups were administrated bleomycin transtracheally. Three kinds of ESCs were administrated to the mice in three different ESC-treat groups respectively one hour after bleomycin exposure. 3.The life-span and hydrocyproline contents were observed. The pathologic changes of the lung and the engraftment of the ESCs in the injured lung were observed. III Twice transplantations of S8ESC to the lung injured by bleomycin. 1. Thirty female C57BL/6J mice were derived into three groups as model group with bleomycin, 1st group (model group with single transplantation of S8ESC) and 2nd group (model group with twice transplantation of S8ESC). 2. One hour after administered to bleomycin transtracheally, the experiment groups were transplanted S8ESC, while the control group was given 0.9% sodium chloride solution. And three days later, 2nd group was transplanted S8ESC for the second time, while 1st group and the control group were given 0.9% sodium chloride solution. 3. The life-span and hydrocyproline contents were examined. The pathologic changes of the lung were observed.Results I. 8mg/kg of bleomycin is the best choice for this research. II. 1. The death rate of the three different ESC-treat groups declined more obviously than that of the control group during 8 days after bleomycin exposure (bleomycin model group 50%, C57BL/6J-ESC group 37.5%, S8-ESC group 20%, human-ESC group 20%). 2. The extent of pathologic changes of the lung in S8-ESC group were slighter significantly than that of the bleomycin model group, but in C57BL/6J-ESC group and human-ESC group, their pathologic changes were similar to that of bleomycin model group. 3. The hydrocyproline contents in S8-ESC group was lower distinctly than that in bleomycin model group (P=0.098,<0.01); the hydrocyproline contents in human-ESC group was also lower than that in bleomycin model group(P=0.0402,<0.05, but >0.01,); there was no significant difference between C57BL/6J-ESC group and bleomycin model group (P=0.627,>0.05). 4. The positive signal of three kinds of ESCs could be found in the lung 1, 3, 12 and 24 hours after the stem cells were administrated, but the positive signal in their lungs was the strongest in 3 hours after stem cells were given. All of the positive signal disappeared three days later. III. 1. The death rate of 2nd group in 14 days and 28 days were lower than the control group and 1st group (2nd group, 70% and 90%; the control group and 1st group, 100%, 100%). 2 The extent of pathologic changes of the lungs in 2nd were slighter than the other groups, and the changes in1st group were slighter than the control group. 3 The hydrocyproline contents in 2nd group is lower than the control group (P=0.004, <0.01) and 1st group. (P=0.048, <0.05)Concluding So we concluded that trasnplatation of S8ESC could increase the tolerance of C57BL/6J mice to bleomycin and decreace the lung injury.