【摘要】：目的：觀察人臍帶間充質(zhì)干細(xì)胞在急性腎小管壞死模型的體內(nèi)的分布、歸巢及再生修復(fù)的作用。 方法： 1、人臍帶間充質(zhì)干細(xì)胞體外培養(yǎng)、擴(kuò)增和分化,移植前均用DAPI標(biāo)記； 2、急性腎小管壞死模型的建立：肌注新鮮配制的0.2%二氯化汞溶液7ml/kg建立急性腎小管壞死的家犬模型；將總量900mg/kg慶大霉素,首次300mg/kg,12h后200mg/kg,余400mg/kg分兩次每隔24h皮下注射,共3天,建立急性腎小管壞死的大鼠模型, 3、人臍帶間充質(zhì)干細(xì)胞移植：采用經(jīng)外周靜脈注射法移植和股動(dòng)脈介入經(jīng)左腎動(dòng)脈直接注射方法移植 4、指標(biāo)測(cè)定：①細(xì)胞定位測(cè)定：采用熒光顯微鏡觀察腎臟DAPI陽(yáng)性細(xì)胞。②腎小管功能檢測(cè)：BUN、SCr、尿蛋白等。③腎臟病理檢測(cè)：HE, PAS染色；④相關(guān)因子的檢測(cè)：免疫組化法檢測(cè)腎組織iNOS的表達(dá) 結(jié)果： 1、二氯化汞溶液和慶大霉素可以成功制造急性腎小管壞死模型 2、對(duì)照組實(shí)驗(yàn)動(dòng)物移植人臍帶間充質(zhì)干細(xì)胞后各個(gè)時(shí)間點(diǎn)均可在腎小管檢測(cè)DAPI陰性 3、造模成功的實(shí)驗(yàn)動(dòng)物移植人臍帶間充質(zhì)干細(xì)胞后各個(gè)時(shí)間點(diǎn)均可在腎小管檢測(cè)DAPI陽(yáng)性,而在心臟、肝臟及胰腺檢測(cè)DAPI陰性；移植后實(shí)驗(yàn)動(dòng)物死亡率明顯降低；腎功能明顯改善；腎臟病理明顯減輕；腎臟局部iNOS促炎癥因子下調(diào)。 結(jié)論： 1、人臍帶間充質(zhì)干細(xì)胞移植后可歸巢到損傷的腎小管,降低動(dòng)物死亡率,減輕腎臟病理?yè)p害及改善腎功能,對(duì)正常腎臟無(wú)影響； 2、對(duì)于急性腎小管壞死模型動(dòng)物,人臍帶間充質(zhì)干細(xì)胞選擇性存在于腎臟內(nèi),對(duì)心臟、肝臟及胰腺無(wú)影響。 3、人臍帶間充質(zhì)干細(xì)胞對(duì)腎小管的修復(fù)通過(guò)下調(diào)iNOS表達(dá)。
[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.
【學(xué)位授予單位】：黑龍江中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R329

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