| Objective:1.To observe the transplantation of the ADMSCs in the lung of the MCT-induced PAH rats by using different cell number.2. To observe the transplantation of the ADMSCs in the lung of the MCT-induced PAH rats different intervention time of the cells.Methods: 1.ADMSCs were isolated and cultured from subcutaneous adipose tissue of Sprague-Dawley rats' groin by collagenase type I. Cells were transfected with Ad5-GFP to be a marker to detect after transplantation A single dose of 40mg/kg MCT was given intraperitoneally to induce the PAH. Animals in the ADMSCs-treated group received an injection of 1 ml GFP labeled ADMSCs [different cells number in 1.0 m L DMEM-F12 respectively,] via the left external jugular vein 2 weeks following MCT administration and the Ctrl and PAH groups were injected with 1.0 m L DMEM-F12 in the same manner. Frozen section was used at the end of the observation. All the slices were stained in sky blue and DAPI, observed the transplantation of the ADMSCs in the fluorescence microscope, then the positive GFP-labeled ADMSCs were found in the randomly selected eight view units in the low power field(10×10). Finally we measured the m PAP and counted the number of the cells and the death percentage.2.The experiment contained 2 parts:(1)In the first part,160 male Sprague-Dawley(SD) rats were randomly divided into normal control group(Ctrl1). PAH1 group,ADMSCs treated group(PAH1-A/Ctrl1-A). PAH and ADMSCs groups were administrated a single dose of 40mg/kg MCT by intraperitoneal injection,,Ctrl and Ctrl-A group were given the equal volume of the normal saline.A certain number of ADMSCs(1×10~6/ml) were transplanted into rats through the left external jugular vein at 14 days after MCT injection while Ctrl and PAH group received equal volume of DMEM-F12, the indexes above were observed in the 1,3,7,14,21 days,the number of the observed rats was 8 at the end of the observation.(2) In the second part, 80 male Sprague-Dawley(SD) rats were randomly divided into normal control group(Ctrl2),PAH2 group,ADMSCs treated group(PAH2-A/Ctrl2-A). The way to induce the PAH rats is the same above. The PAH2-A and Ctrl2-A group were divided into 2 subgroups after 2 weeks of the injection of the MCT, the different number of ADMSCs [5×105/ml(Ctrl2-A1/PAH2-A1) ? 1×10~6/ml(Ctrl2-A2/PAH2-A2) ?5×10~6/ml(Ctrl2-A3/PAH2-A3)?1×10~7/ml(Ctrl2-A4/PAH2-A4)] were transplanted into rats at 14 days after MCT injection,while Ctrl2 and PAH2 group received equal volume of DMEM-F12, All these rats were observed 21 days.Results:1. The number of the ADMSCs transplantation in lungs of PAH1-A was time dependent, with the time went, the number of the ADMSCs transplantation in lungs counted less.(the number of the ADMSCs in the 1,3,7,14,21 days is as followed:12.62±1.68,7.75±1.67,6.37±1.18,4.50±1.19,2.50±1.20), The ADMSCs was not seen in the Ctrl1 group. The m PAP was decreased in PAH1-A compared with PAH1.2.The number of the ADMSCs transplantation in lungs of PAH2-A was cell number dependent, with the number of the transplanted cells, the counted cells were increased, especially the in the PAH2-A2 and PAH2-A3 group(PAH2-A2,PAH2-A3:7.25±2.12?13.25±1.91,p<0.05). However, compared with PAH2-A3 group(5×10~6/ml), there were no significant difference in PAH2-A4(10~7/ml) group. The ADMSCs was not observed in the Ctrl2 group. Compared with the PAH2 group,the m PAP of the PAH2-A2, PAH2-A3 and PAH2-A4 group were decreased,while there were no statistical difference in PAH2-A3 and PAH2-A4 group, and the death percentage was increased in PAH2-A4.Conclusions:1. ADMSCs could be transplanted in pulmonary vascular of the MCT-induced PAH rats stably by injected the number of the ADMSCs,ranging from 1×10~6/ml to 5×10~6/ml.2. MPAP could be decreased by injecting the number of ADMSCs after 3 weeks 14 days after intraperitoneal injection of 40mg/kg MCT,transplantation of 1×10~6 ~5×10~6/ml ADMSCs for 21 days could effectively reduce m PAP in rats. |
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