本文選題：腎間質(zhì)纖維化　切入點：骨髓間充質(zhì)干細胞　出處：《昆明醫(yī)學院》2009年碩士論文

【摘要】： 目的 建立兔腎單側(cè)輸尿管結(jié)扎制作腎間質(zhì)纖維化動物模型,將體外分離標記的自體骨髓間充質(zhì)干細胞經(jīng)股動脈插管,腎動脈灌注回輸細胞,觀察移植自體骨髓間充質(zhì)干細胞能否歸巢至損傷腎臟組織,及其在腎組織中的分布,能否促進其再生修復,從而改善腎功能。 方法 ①兔腎間質(zhì)纖維化模型的建立:用單側(cè)輸尿管結(jié)扎建立兔腎間質(zhì)纖維化模型,動態(tài)觀察4周,于造模的第0、1、2、3周末檢測血尿素氮(Bun)、肌酐(Scr)含量及第3周采用單光子發(fā)射電子計算機斷層成像儀(SPECT),掃描監(jiān)測腎功能變化;4周末采用HE染色、PAS染色,觀察腎組織變化。②細胞分離純化:氯化銨破碎紅細胞聯(lián)合貼壁培養(yǎng)法分離培養(yǎng)兔骨髓間充質(zhì)干細胞。③Brdu標記移植細胞:Brdu標記移植骨髓間充質(zhì)干細胞,觀察標記率。④移植細胞:于造模成功后的第4周經(jīng)右股動脈插管行自體骨髓間充質(zhì)干細胞腎動脈灌注回輸細胞,分別在移植前、移植后的第1、3、5、7周行血生化Bun、Scr檢測,腎組織病理學分析、Brdu免疫組化分析。 結(jié)果 ①動物模型的血生化指標、病理組織學變化及SPECT監(jiān)測:造模前兩組血Bun、Scr值沒有顯著性差異(P＞0.05)。于造模后的第1周起兩組血Bun、Scr逐漸升高,第3周達到最高水平,有顯著差異性,與對照組比較有統(tǒng)計學意義(P＜0.05)。SPECT顯示患側(cè)腎明顯變形,腎顯影變淡,呈新月形,腎小球濾過率明顯低于對照組,模型組兔腎間質(zhì)纖維增生,呈纖維化改變,炎癥細胞浸潤,腎小管上皮部分脫落,濁腫,部分腎小管萎縮,腎小球有出血,其余未見明顯異常。②移植細胞的標記:以30umol/L終濃度的Brdu孵育骨髓間充質(zhì)干細胞72小時的陽性標記率均達95%以上,而標記組細胞形態(tài)、生長和增殖與對照組未見明顯異常。③移植后腎功能檢測:移植后第3周起血Bun、Scr逐漸降低,與造模后及對照組比較有統(tǒng)計學意義(P＜0.05)。④移植后腎病理組織學變化:移植后第1周所見腎小管上皮細胞變性、壞死脫落,腔內(nèi)可見透明管型,在移植的第3周、第5周可見腎小管上皮細胞混濁腫脹,隨著時間延長,壞死腎小管逐漸修復。⑤腎組織Brdu免疫組化分析:移植組于移植后的第1周逐漸顯示腎小管和部分腎小球,Brdu陽性染色持續(xù)至實驗結(jié)束,陽性染色主要分布于腎組織外的髓質(zhì)和皮髓交界處,部分腎小球也呈陽性染色,而對照組Brdu染色則呈陰性。 結(jié)論 ①通過病理學及SPECT監(jiān)測,單側(cè)輸尿管結(jié)扎制作該模型具有明顯的腎間質(zhì)纖維化特征,為臨床實驗研究提供了實驗模型。②經(jīng)腎動脈灌注途徑移植的骨髓間充質(zhì)干細胞能遷移并定居于腎臟,參與了損傷腎臟結(jié)構(gòu)的修復。③兔腎間質(zhì)纖維化后,移植的自體骨髓間充質(zhì)干細胞對血Bun和Scr的升高有一定的抑制作用,促進了腎臟功能的恢復。④骨髓間充質(zhì)干細胞移植后第1周后即可進入腎臟,主要分布于腎臟的外髓區(qū)。
[Abstract]:PurposeAn animal model of renal interstitial fibrosis was established by unilateral ureteral ligation in rabbits. Autologous bone marrow-derived mesenchymal stem cells (MSCs) isolated and labeled in vitro were intubated through femoral artery and injected back into renal artery.To observe whether autologous bone marrow mesenchymal stem cells (BMSCs) can homing to the injured kidney tissue and its distribution in the renal tissue, and whether it can promote its regeneration and repair, thereby improving the renal function.Method1 Establishment of rabbit renal interstitial fibrosis model: rabbit renal interstitial fibrosis model was established by unilateral ureteral ligation and observed dynamically for 4 weeks.Blood urea nitrogen (bun) and creatinine (SCR) contents were detected at the end of the 3rd week of the model, and SPECTX was used to detect the changes of renal function by single photon emission computed tomography (SPECTX) at the 3rd week. The changes of renal function were detected by HE staining and pas staining at the end of 4 weeks.Observe the renal tissue changes .2 cell isolation and purification: ammonium chloride broken red blood cells combined with adherent culture method to isolate and culture rabbit bone marrow mesenchymal stem cells. 3 Brdu labeled transplantation cells labeled bone marrow mesenchymal stem cells labeled with Brdu labeled bone marrow mesenchymal stem cells.To observe the labeling rate of transplanted cells: the renal artery infusion of autologous bone marrow mesenchymal stem cells was performed through right femoral artery catheterization at the 4th week after the model was successfully established, and the blood biochemistry Bunn SCR was detected before transplantation and 57 weeks after transplantation, respectively.Renal histopathology and Brdu immunohistochemical analysis.Result1Blood biochemical indexes, histopathological changes and SPECT monitoring of the animal model: there was no significant difference between the two groups before the establishment of the model (P > 0.05).There was no obvious abnormal renal function in the growth and proliferation group. 3. The blood BunScr level decreased gradually from the third week after transplantation.At the 5th week, the tubule epithelial cells were turbid and swollen, and the tubuloepithelial cells showed turbid swelling over time.The positive staining was mainly located at the junction of medulla and dermis outside renal tissue, and some glomeruli were also positive staining, but Brdu staining was negative in the control group.Conclusion1 by pathological and SPECT monitoring, the model of unilateral ureteral ligation had obvious characteristics of renal interstitial fibrosis.The experimental model was provided for the clinical study. 2 the bone marrow mesenchymal stem cells transplanted by renal artery perfusion were able to migrate and settle in the kidney and participated in the repair of renal interstitial fibrosis in rabbits.Autologous bone marrow mesenchymal stem cells (BMSCs) could inhibit the increase of Bun and Scr and promote the recovery of renal function. 4. BMSCs could enter the kidney 1 week after transplantation, mainly distributed in the extramedullary area of the kidney.
【學位授予單位】：昆明醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R692.9;R-332

【參考文獻】

相關(guān)期刊論文 前6條

1 傅曉晴,武一曼,陳振彬,陳華欽,陳琪,陳惠穎,顧云龍;腺嘌呤制作腎陽虛型慢性腎功能衰竭大鼠模型的電鏡病理學研究[J];福建中醫(yī)學院學報;2002年03期

2 周小舟,張盛光,陽曉,魏毅;腺嘌呤所致大鼠慢性腎功能衰竭的機理研究[J];基礎醫(yī)學與臨床;1997年01期

3 張靚,唐朝樞;第四屆國際松弛素及其相關(guān)肽會議在美召開[J];生理科學進展;2005年01期

4 王勁,羅成基,郭朝華,張勇;間充質(zhì)干細胞與相關(guān)因子[J];中國實驗血液學雜志;2002年05期

5 鄭法雷;慢性腎功能衰竭進展的機制與防治[J];中華內(nèi)科雜志;1998年03期

6 傅鵬,崔若蘭,袁偉杰,戴建新,孫樹漢;缺血性急性腎功能衰竭病程進展中p27~(kip1)作用的研究[J];中華腎臟病雜志;2002年04期

，

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