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作 者:宁昕杰[1] 王辉[1] 陆新华[1] 罗骏成 巴越洋[2]
机构地区:[1]中山大学附属第三医院神经外科,广东广州510630 [2]中山市人民医院神经外科,广东中山520483
出 处:《中国病理生理杂志》2015年第3期392-396,共5页Chinese Journal of Pathophysiology
基 金:国家自然科学基金资助项目(No.30901542);广东省自然科学基金资助项目(No.7301217);广东省医学科学基金资助项目(No.B2007061);广东省科技计划(No.2012B031800056);中山大学高校基本业务费青年教师培育资助项目(No.12ykpy44)
摘 要:目的:探讨神经损伤后microRNA-21(miR-21)促进雪旺细胞增殖的分子机制。方法:通过实时荧光定量PCR检测miR-21的表达。通过脂质体介导将人工合成的miR-21模拟物(mimic)及其阴性对照转染大鼠雪旺细胞,CCK-8法检测转染miR-21的雪旺细胞的增殖。流式细胞术分析miR-21对雪旺细胞细胞周期的影响。使用Western blotting实验分析转化生长因子β诱导蛋白(TGFBI)和cyclin D1的表达水平。结果:miR-21在损伤模型组中的表达分别为假手术组和正常神经组织的(7.87±0.75)和(7.75±0.80)倍(P<0.01)。miR-21 mimic组miR-21的表达分别为对照组和空白组的(2.21±0.14)和(2.29±0.21)倍(P<0.05)。转染48 h后miR-21 mimic组CCK-8实验的A450值较阴性对照组和空白对照组增高(P<0.05)。实验组细胞增殖指数高于阴性对照组和空白对照组(P<0.01)。同时与对照组相比,转染miR-21后48 h实验组细胞TGFBI明显降低,cyclin D1表达增多(P<0.05)。结论:miR-21可促进雪旺细胞增殖,其机制可能与其下调TGFBI的表达有关。AIM: To investigate the molecular mechanism of microRNA-21 (miR-21) in the regulation of Schwann cell proliferation following nerve injury.METHODS:The expression of miR-2l was detected by real-time PCR. Synthetic miR-21 mimic and its control were transfected into rat Schwann cells.CCK-8 assay was performed to evaluate the influence of miR-21 on the proliferation of Schwann cells.The cell cycle distribution was determined by flow cytometry. The expression of transforming growth factorβ-induced protein ( TGFBI) and cyclin D1 were detected by Western blotting. RESULTS:The expression of miR-21 in model group was 7.87 ±0.75 and 7.75 ±0.80 times higher than that in sham operation group and blank group respectively.After transfected with miR-21 mimic, the expression of miR-21 in experimen-tal group was 2.21 ±0.14 and 2.29 ±0.21 times higher than that in negative control group and blank group respectively. Moreover, the A450 value of CCK-8 assay in experimental group at 48 h was higher than that in negative control group and blank group.The proliferation index in experimental group was higher than that in negative control group and blank group. At the same time, the expression of TGFBI obviously decreased and the cyclin D1 increased in the Schwann cells 48 h after transfection with miR-21.CONCLUSION:miR-21 promotes the proliferation activity of Schwann cells by down-regulating TGFBI expression.
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