【摘 要】
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目的:调控胰岛早期凋亡是胰岛移植成功的关键.本研究旨在观察BMSCs与胰岛共移植对胰岛早期凋亡的影响,并初步探讨其可能作用机理.方法:(1)体内实验:全骨髓贴壁法分离培养BALB/c乳鼠骨髓间充质干细胞(BMSCs)并鉴定.胶原酶消化法获取供体胰岛并计数.腹腔注射STZ建立糖尿病BALB/c小鼠模型.造模成功后,将小鼠随机分配到4个实验组:①胰岛单独移植组:肾包膜下注射胰岛350个(n=10),②
【机 构】
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中南大学湘雅医院内分泌科 长沙410008 华中科技大学同济医学院附属同济医院器官移植研究所 武汉
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目的:调控胰岛早期凋亡是胰岛移植成功的关键.本研究旨在观察BMSCs与胰岛共移植对胰岛早期凋亡的影响,并初步探讨其可能作用机理.方法:(1)体内实验:全骨髓贴壁法分离培养BALB/c乳鼠骨髓间充质干细胞(BMSCs)并鉴定.胶原酶消化法获取供体胰岛并计数.腹腔注射STZ建立糖尿病BALB/c小鼠模型.造模成功后,将小鼠随机分配到4个实验组:①胰岛单独移植组:肾包膜下注射胰岛350个(n=10),②胰岛+BMSCs共移植组:350个胰岛+7×105BMSCs(n=10),③BMSCs单独移植组:7×105 BMSCs(n=6),④假手术对照组:等量RMPI1640液(n=6).观察术后血糖变化,并以TUNEL+胰岛素检测术后1、3、7、28胰岛B凋亡状况;透视电镜观察胰岛B细胞内质网结构改变,并以免疫荧光法观察胰岛B细胞内质网应激相关指标BIP, CHOP的表达.(2)体外实验:分为胰岛单独培养组和胰岛+BMSCs共培养组.分别于培养第1、3、5、7、14天以AO/PI染色法观察两组胰岛存活死亡情况;GSIS(葡萄糖刺激胰岛素分泌)法检测胰岛分泌功能;并以real-time PCR、western blot检测胰岛细胞内质网应激相关指标BIP,CHOP, caspase12的RNA及蛋白质表达.
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