下颌下腺干-组细胞论文：不同方法诱导SD大鼠下颌下腺干-祖细胞增殖研究

下颌下腺干-组细胞论文：不同诱导SD大鼠下颌下腺干-祖细胞增殖研究 下颌下腺干/组细胞论文:不同方法诱导SD大鼠下颌下腺干/祖细胞增殖研究 【中文摘要】比较SD大鼠下颌下腺导管结扎和热环境处理后腺体内干/祖细胞的增殖情况,寻找能更好诱导腺体内干/祖细胞增殖的方法。方法:SD雄性大鼠15只,8周龄大小,大鼠单纯随机抽样分为正常组、结扎组和加热组三组,每组5只。结扎组大鼠麻醉下行双侧下颌下腺导管结扎,饲养6天后处死并摘除腺体。加热组大鼠置于34.5?0.5?环境饲养,48小时后处死并摘除下颌下腺腺体。正常组大鼠按正常环境给予饲养后摘除下颌下腺腺体。各组摘除的腺体行形态学、HE染色及CD117、Laminin、整合蛋白α6β1、ki-67免组织化学观察,流式细胞分析整合蛋白α6β1、c-Kit,CD90.1阳性细胞含量。结果:形态学观察:三组腺体中结扎组腺体体积最小,包膜不完整,质地较韧;加热组和正常组形态上相近,包膜完整,质地柔软,但加热组腺体体积较正常组大。腺体重量与大鼠体重之比发现:加热组比正常组大25.3%(p<0.05);正常组比结扎组大29.3%(p<0.05);加热组比结扎组大47.2%(p<0.05)。HE染色组织切片中导管结扎组结构紊乱,腺泡萎缩,导管增殖,间质纤维成分较多;加热组腺体腺泡导管结构清晰,与正常组相似,但高倍镜下可见有丝分裂相。流式细胞检测下颌下腺腺体内整合蛋白α。β,阳性细胞率在加热组为26.2?2.4%,结扎组为16.9?2.6%,正常组为10.6?0.7%,统计学分析在加热组中整合蛋白α。β1阳性细胞表达率分别较结扎组和正常组高(p<0.05), 而结扎组整合蛋白α。β1阳性细胞表达率较正常组高(p<0.05): CD90.1阳性细胞率在加热组为33.0?5.4%,结扎组为25.0?4.1%,正 常组为18.1?3.4%,统计学分析在加热组中CD90.1阳性细胞表达率 分别较结扎组和正常组高(p<0.05),结扎组CD90.1阳性细胞表达率 较正常组高(p<0.05); c-Kit阳性细胞率在加热组为13.4?3.4%,结 扎组为13.8?3.0%,正常组为8.5?3.1%,统计学分析显示在加热组 中c-Kit阳性细胞表达率与结扎组无统计学差异(p>0.05),但在加热 组和结扎组中c-Kit阳性细胞表达率均较正常组高(p<0.05)结论:加 热环境比导管结扎更能有效刺激涎腺腺体内表达整合蛋白α。β1和 CD90.1阳性的细胞增殖。加热环境和导管结扎两组对涎腺腺体内表 达c-Kit阳性细胞的增殖没有明显差异。 【英文摘要】:In order to find an efficient way to induce proliferation of stem/progenitor cell in salivary gland, we compared the proliferation ability of salivary stem/progenitor cells between main duct ligation and heat stress on submandibular gland of SD rat.Methods:Fifteen male Sprague-Dawlye(SD)Rats,10-weeks-old, divide into heat, ligation and normal groups, randomly. Under anesthesia, the duct of glands were ligated and then the glands were harvested after six days. The heat group rat were placed in a heat acclimation box with an ambient temperature of 34.5?for 48 hours and then the glands were harvested. The normal group rats were fed in normal environment and then removed the submandibular gland. All gland were tested in morphology and detected the percentage of cells that expressed stem/progenitor cell surface marker integrinα6β1, CD90.1, c-Kit with Flow Cytometry, respectively. (FCM)Results:Morphology observation, the glands volume was smallest in the duct ligation group, the gland shape of duct ligation changed to spindle-like and incomplete capsule. The shape of gland in the heat group was similar to normal group, and the color was slightly rosy. The gland weight to body weight ratio in the heat group was 25.3% larger than normal group and was 47.2% larger than duct ligation group. The gland weight to body weight ratio in the normal group was 29.3% larger than duct ligation group. Histological observation, HE and immunohistochemical staining:under the light microscope, the acinar and duct of glands in the heat and normal groups could be observed clearly, but the mitotic figures could be seen in the heat group. After duct ligation, acinar cells disappear and proliferation of duct could be occured. Flow cytometric analysis demonstrated that the percentage of cells positive forα6β1 integrin in the heat group was 26.2?2.4%, the ligation group was 16.9?2.6%, and the normal group was 10.6?0.7%. Statistic demonstrated that the the percentage of cells expressedα6β1integrin in the heat group was highest than duct ligation and normal groups (p<0.05), and the percentage of cells expressedα6β1integrin in the duct ligation group was higher than normal group (p<0.05). The percentage of cells positive for CD90.1 in the heat group, ligation group and normal group were 33.0?25.4%,25.0?4.1%,18.1?3.4% respectively. Statistic demonstrated that the the percentage of cells expressed CD90.1 in the heat group was highest than duct ligation and normal groups (p<0.05), and the percentage of cells expressed CD90.1 in the duct ligation group was higher than normal group (p<0.05). The percentage of cells expressed c-Kit in the heat group was 13.4?3.4%, in the duct ligation group was 13.8?3.0% and in normal group was 8.5?3.1%. Statistic demonstrated that the the percentage of cells expressed c-Kit had no statistic difference between heat group and duct ligation group (p>0.05), but the percentage of cells expressed c-Kit in heat group and duct ligation were higher than normal group(P<0.05).Conclusion:The heat-condition could more useful to induce the proliferation of cells that expressedα6β1integrin and CD90.1. And there was similar effect to induce the proliferation of cells that expressed c-Kit between the heat-condition and duct ligation groups. 【关键词】下颌下腺干/组细胞 增殖 热处理 导管结扎 【备注】索购全文在线加好友:1.3.9.9.3.8848 同时提供论文写作一对一指导和论文发表委托服务 【英文关键词】Submandibular gland stem/progenitor cells heat condition duct ligation proliferation 【目录】不同方法诱导SD大鼠下颌下腺干/祖细胞增殖研究 英文缩略词 4-5 目录 5-7 中文摘要 7-9 Abstract 9-10 前言 11-13 试验 一:Sprague-Dawlye大鼠下颌下腺主导管结扎组织损伤模型及热处理模型的建立 13-18 实验材料 13 实验方法 13-14 结果 14-16 讨论 16-17 参考文献 17-18 实验二:流式细胞检测正常组、结扎组、加热组腺体内干/祖细胞含量 18-27 实验材料 18-19 实验方法 19-20 结果 20-23 讨论 23-25 结论 25-26 参考文献 26-27 实验三:SD大鼠下颌下腺组织学变化 27-38 实验材料 27-28 实验方法 28-30 实验结果 30-35 讨论 35-37 参考文献 37-38 全文总结 38-40 综述:人工涎腺组织工程的研究进展 40-52 参考文献 47-52 致谢 52-53 作者简介 53