The Differentiation Of Bone Marrow Stem Cells In The Microenvironment Of Glomerulosclerosis

Objective To investigate the differentiation of bone marrow stem cells in the microenvironment of glomerulosclerosis .And to explore the progress of chronic kidney disease and the mechanisms of glomerulosclerosis .Methods Transgenic C57BL/6 male mice labeled with enhanced green flouresent protein(EGFP) were bone marrow donors, 80 C57BL/6 female mice without fluorescence labelces, recipients. Female mice accepted bone marrow mononuclear cells labeled EGFP after the total body homogeneous radiation. Four weeks after the bone marrow transplantation, female mice were randomly divided into two groups:(1)Bone marrow transplantation plus Glomerulosclerosis group(BMSC+GS Group): 40 female mice were carried out unilateral nephrectomy and injected with 5mg/kg and 3mg/kg of adriamycin via tail vein at 7,28 days after nephrectomy ;( 2 ) Bone marrow transplantation plus sham operation group(BMSC+Sham Group): 40 female mice were carried out sham operation and 0.9% Sodium Chloride solution injection of equal volume. 8 C57BL/6 female mice without any treatment were as Control Group. 2,4,8,12,16 weeks after nephrectomy ,eight mice randomly selected from two groups of marrow transplantation were sacrificed.All mice of the Control Group were sacrificed at the 16 weeks.Indicators were detected as follow:(1)Urine protein,blood urea nitrogen and Creatinine;(2)Renal pathological changes were observed by light microscopy;(3)The EGFP positive cellsˊdesposition in renal was observed by fluorescence histochemistry and immunohistochemistry;(4)The differentiation of bone marrow stem cells was observed through double labeled fluorescence histochemistry method by laser confocal microscopy.Result (1)Urine protein were negative in Control Group and BMSC+Sham Group at every time point. Those in BMSC+GS Group were weak positive in 2 weeks,and then were positive with time;Compared with Control Group and BMSC+Sham Group,the BUN and Cr were increased (p<0.05);(2)Renal pathological changes:No Renal pathological changes were found at any time in Control Group and BMSC+Sham Group.Mesangial proliferation was observed at 4 weeks and the focal segmental glomerular sclerosis was observed at 12 and 16 weeks in BMSC+GS Group;(3)A little EGFP positive cells were observed in glomerular,tubular and interstitial areas at every time point in BMSC+Sham Group.For BMSC+GS Group,many EGFP positive cells were observed in glomerular at 2 and 4 weeks,then the positive cells became more in tubular and interstitial areas(;4)For BMSC+GS Group,EGFP/α-SMA double positive cells were observed in glomerular and interstitia at 2 weeks and became more and more with time by laser confocal microscopy.Conclusion (1)The bone marrow stem cells can home to the injured kidneys in the microenvironment of glomerulosclerosis;(2)The bone marrow stem cells can transfer to renal parenchymal cells such as glomerular mesangial cells,epithelial cells,endothelial cells and renal tubular epithelial cells;The bone marrow stem cells can also transfer to matri-secreting cells and take participate the process of glomerulosclerosis and interstitial fibrosis in the microenvironment of glomerulosclerosis.