| Kidney is one of sensitive organs in ischemia-reperfusion injury. Clinically, shock, disseminated intravascular coagulation, extracorporeal shock wave lithotripsy, kidney trauma surgery and kidney transplants inevitably result in renal ischemia-reperfusion injury. The effect of clinical drugs is so poor for renal ischemia-reperfusion injury that it develops chronic renal failure. Over the last decade, there is no significantly effective therapy to improve acute renal failure caused by renal ischemia-reperfusion injury. Stem cell research is hot in life sciences. The results of stem cell research were appraised as one of " the top 10" scientific and technological progress in the world by "Science" magazine in 1999, 2000 and 2003 and is one of most attractive scientific and technological achievements in the 21st century. Bone marrow mesenchymal stem cells (BMSCs) are a kind of adult stem cells and have become one of the most important seeded cells for tissue engineering. Mesenchymal stem cells have rich source and are easily obtained. Mesenchymal stem cells not only are capable of self-renewal and multi-differentiation, but also have obvious characteristics of plasticity. BMSCs which maintain the physiological states of hematopoietic stem cell for long term as an important component in hematopoietic microenvironment can differentiate into osteoblasts, cartilage and adipocyte and the cell types of the other tissue in mesoderm, such as muscle cells under the appropriate circumstances. Further, BMSCs even transdifferentiate into the cell types of the other tissues originated from ectoderm and endoderm, including neuron-like cells, glial cells, epidermic cells and hepatocyte, etc. Plasticity of BMSCs suggests that the treatment of replacing injured renal parenchymal cells with BMSCs may be effective.In this study, we cultured and expanded BMSCs first; then BMSCs were transplanted into ischemia-reperfusion kidneys of inbreed strain mice via the tail vein. A series of indexes were detected to observe that BMSCs can promote the repair of injured kidney. The repair mechanism of renal injury was explored through observating the trace of DAPI-labeling donator's BMSCs in acceptor's body. The study will provide a new method of the treatment of renal ischemia-reperfusion injury and contribute to the study of the treatment of kidney diseases by cells.Objective: To research the growth characteristics of BMSCs, establish the normative mice model of renal ischemia-reperfusion injury and observe the repair effect of the transplantation of BMSCs on renal injury. Our aim also includes observing the differentiation of BMSCs into renal tubular epithelial cells and exploring the repair mechanism of renal ischemia-reperfusion injury by BMSCs.Methods: BMSCs were separated by combing density gradient centrifugation with adherent culture from bone marrow cell suspension of mice. Then BMSCs were transplanted into ischemia-reperfusion kidneys via the tail vein. Renal function, the indexes of renal oxidative stress, morphologic changes and renal pathology changes were detected. The mechanism of the repair of renal injury was explored through observation of the trace of DAPI-labeling donator's BMSCs in acceptor's body.Results: 1. BMSCs had a characteristic of strong adherence and grew like colony in primary culture. BMSCs appeared uniform long fusiform shapes after passaged. The homogeneity of BMSCs was markedly improved in serial subcultivation.2. BMSCs could differentiate into osteoblast and adipocyte by culturing in induced media of osteoblast differentiation and adipocyte differentiation. There was brownish-black sediment in the osteoblast by alkaline phosphatase staining. There was red sediment in the adipocyte by oil red O staining.3. The renal ischemia-reperfusion injury was repaired after transplantation of BMSCs into C57BL/6J mice. Transplanted mice appeared the good state. Indexes of renal function and renal histomorphology had been improved. The level of indexes of oxidative stress declined.4. The cells with blue fluorescent were found in C57BL/6J mice kidney frozen section. It showed that donator's BMSCs participated in the reparation of injured renal tissue.5. After transplantation of BMSCs into ICR mice, renal ischemia-reperfusion injury was not improved. Mice appeared the poor state. Indexes of renal function and oxidative stress were not different from the ones in the control group. Some indexes were significantly higher than those of the control group. Renal pathological changes were not improved. Conclusion: 1. In this study, BMSCs were generously obtained from bone marrow cell suspension of mice by combining density gradient centrifugation with adherent culture. BMSCs not only had a high purification, homogeneity and stability, but also could unlimited proliferation and multi-differentiation.2. The renal ischemia-reperfusion injury model was successful established. The recovery of renal ischemia- reperfusion injury in the host were improved after transplantation of BMSCs into C57BL/6J mice by tail vain and the extent of renal injury was improved. Mice had the good state. Indexes of renal function, oxidative stress and renal pathology had been improved.3. The study of traces of BMSCs in the host body confirmed BMSCs could home to damage tissue. And firstly, BMSCs repair the damaged vascular endothelial tissue, and then participate in repairing renal injury.4. The further experiment need to be studied on the mechanisms that BMSCs could repair renal ischemia-reperfusion injury in uninbred animals, but they can't repair renal ischemia-reperfusion injury in inbred animals.
|
PDF Full Text Request