人脐带间充质干细胞移植对急性肾小管坏死再生修复的实验研究

发布时间：2018-11-28 14:20

【摘要】：目的：观察人脐带间充质干细胞在急性肾小管坏死模型的体内的分布、归巢及再生修复的作用。方法： 1、人脐带间充质干细胞体外培养、扩增和分化,移植前均用DAPI标记； 2、急性肾小管坏死模型的建立：肌注新鲜配制的0.2%二氯化汞溶液7ml/kg建立急性肾小管坏死的家犬模型；将总量900mg/kg庆大霉素,首次300mg/kg,12h后200mg/kg,余400mg/kg分两次每隔24h皮下注射,共3天,建立急性肾小管坏死的大鼠模型, 3、人脐带间充质干细胞移植：采用经外周静脉注射法移植和股动脉介入经左肾动脉直接注射方法移植 4、指标测定：①细胞定位测定：采用荧光显微镜观察肾脏DAPI阳性细胞。②肾小管功能检测：BUN、SCr、尿蛋白等。③肾脏病理检测：HE, PAS染色；④相关因子的检测：免疫组化法检测肾组织iNOS的表达结果： 1、二氯化汞溶液和庆大霉素可以成功制造急性肾小管坏死模型 2、对照组实验动物移植人脐带间充质干细胞后各个时间点均可在肾小管检测DAPI阴性 3、造模成功的实验动物移植人脐带间充质干细胞后各个时间点均可在肾小管检测DAPI阳性,而在心脏、肝脏及胰腺检测DAPI阴性；移植后实验动物死亡率明显降低；肾功能明显改善；肾脏病理明显减轻；肾脏局部iNOS促炎症因子下调。结论： 1、人脐带间充质干细胞移植后可归巢到损伤的肾小管,降低动物死亡率,减轻肾脏病理损害及改善肾功能,对正常肾脏无影响； 2、对于急性肾小管坏死模型动物,人脐带间充质干细胞选择性存在于肾脏内,对心脏、肝脏及胰腺无影响。 3、人脐带间充质干细胞对肾小管的修复通过下调iNOS表达。
[Abstract]:Aim: to observe the distribution of human umbilical cord mesenchymal stem cells in acute renal tubular necrosis model and the effects of homing and regeneration. Methods: 1. Human umbilical cord mesenchymal stem cells were cultured, amplified and differentiated in vitro, and were labeled with DAPI before transplantation. (2) Establishment of acute tubular necrosis model: the canine model of acute tubular necrosis was established by intramuscular injection of fresh 0.2% mercuric chloride solution (7ml/kg). The total amount of 900mg/kg gentamicin, 200mg / kg after the first 300mg / kg of Gentamicin, was subcutaneously injected twice every 24 hours for 3 days to establish a rat model of acute tubular necrosis. Transplantation of human umbilical cord mesenchymal stem cells (HMSCs) was performed by peripheral vein injection and femoral artery interventional direct injection via left renal artery. Index determination: 1 Cell localization: DAPI positive cells in kidney were observed by fluorescence microscope. 2 Renal tubule function: BUN,SCr, urine protein, etc. 3: HE, PAS staining was detected by renal pathology. 4 Detection of related factors: immunohistochemical method was used to detect the expression of iNOS in renal tissue. 1. Acute tubular necrosis model was successfully made by mercuric chloride solution and gentamicin. In control group, human umbilical cord mesenchymal stem cells were detected for DAPI negative 3 at all time points after transplantation of human umbilical cord mesenchymal stem cells, and DAPI positive in renal tubules at all time points after transplantation of human umbilical cord mesenchymal stem cells from successful experimental animals. DAPI was negative in heart, liver and pancreas. After transplantation, the mortality of experimental animals was significantly reduced, renal function was improved, renal pathology was alleviated, and local iNOS pro-inflammatory factor was down-regulated. Conclusion: 1. Human umbilical cord mesenchymal stem cells can homing to damaged renal tubules after transplantation, which can reduce the mortality of animals, alleviate renal pathological damage and improve renal function, and have no effect on normal kidneys. 2. In acute renal tubular necrosis model, human umbilical cord mesenchymal stem cells were selectively present in the kidney and had no effect on heart, liver and pancreas. 3. The repair of renal tubules by human umbilical cord mesenchymal stem cells down-regulated the expression of iNOS.
【学位授予单位】：黑龙江中医药大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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