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机构地区:[1]南方医科大学肿瘤研究所,广州510515 [2]南方医科大学组织胚胎教研室,广州510515
出 处:《中国比较医学杂志》2014年第1期35-39,I0009,共6页Chinese Journal of Comparative Medicine
基 金:国家自然科学基金委;广东省政府联合重点基金(U0732006)
摘 要:目的探讨巢蛋白(nestin)基因过表达对小鼠部分肝切除后肝再生增殖能力的影响及其可能的分子机制。方法应用荧光定量PCR、Western blot和免疫组化的方法检测转基因小鼠肝脏中外源基因nestin的表达情况。使用Brdu标记法比较转基因小鼠和野生型小鼠肝脏在不同状态下(静息状态、肝叶切除后24 h、肝叶切除后72 h)肝细胞增殖能力的差异。结果荧光定量PCR、Western blot和免疫组化证实转基因小鼠肝脏中存在nestin的高表达状态。在静息状态下,野生鼠和nestin过表达的转基因鼠肝脏BrdU标记率几乎为零,两者无明显差异;但在肝大部切除的情况下,nestin过表达的转基因鼠有更多的细胞进入增殖状态,增殖能力较野生鼠明显增强,差别具有统计学意义(24 h:t=2.996,P=0.024;72 h:t=2.915,P=0.027)。结论 nestin过表达对静息状态下的肝脏增殖无影响,但可增加损伤修复情况下肝脏的再生增殖能力。表明nestin过表达对静止期细胞作用不明显,但在一些病理或刺激状态下nestin可以通过促进DNA的合成而加速细胞的分裂增殖。Objective To study the influence and possible mechanism of nestin overexpression on the partial hepatectomy regeneration and proliferation capacity in mice. Methods In order to confirm the overexpression status,the expression levels of nestin in liver was detected by fluorescence quantitative PCR,Western blot and immunohistoehemistry. Then,The difference of proliferation index in wild type mice and transgenic mice liver was detected by Brdu label,the mice were divided into 3 groups( in the resting state,24hours later of liver subtotal nephrectomy,72hours later of liver subtotal nephrectomy). Results Fluorescence quantitative PCR,Western blot and immunohistochemistry confirmed overexpression of nestin in the liver of the transgenic mice. In the resting state, Brdu labelling index is almost zero both in the transgenic mice and the wild type mice, showing no difference between them;However,in the case of partial hepateetomy,more cells in the transgenic mice enter into proliferation status, suggesting that overexpression of nestin enhanced the regeneration and proliferation capacity of the liver cells in the transgenic mice when compared to the wild type (24hours later:t = 2. 996 ,P = 0. 024 ;72hours later : t = 2.915, P = 0. 027 ). Conclusion Nestin overexpression had no effect on the liver proliferation inthe resting state,but can increase the proliferative capacity in the case of partial hepatectomy ( PH). This indicate that Nestin overexpression had no obvious effect on quiescent cell,but in some pathological changes or stimulus state,nestin can increase the proliferation of cells through promote the formation of DNA.
分 类 号:R332[医药卫生—人体生理学]
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