【摘要】 目的:探讨细胞周期素E表达与人肾间质成纤维细胞体外增殖的关系。方法:实验于2004-06/2005-10在泸州医学院分子病理实验室和中心实验室完成。选取第3代对数生长期人肾间质成纤维细胞用于实验。采用组织块培养法获取人肾间质成纤维细胞并鉴定;用免疫细胞化学SABC法和图像分析技术检测体外培养第1,3,5,7,9天人肾间质成纤维细胞的细胞数量、细胞周期素E表达阳性的细胞数量以及阳性细胞的平均吸光度值。各时间点阳性细胞细胞周期素E平均吸光度值之间的两两比较采用q检验。结果:①体外培养1～9d,人肾间质成纤维细胞细胞总量逐渐增加,生长曲线呈“S”形,第3～5天细胞数量增长最迅速为对数生长期,第7～9天细胞数量增长最平缓。②细胞周期素E表达阳性细胞比率以第3,5天最高,随后阳性细胞的实际数量仍有所增加,而阳性比率开始缓慢降低。③细胞周期素E平均吸光度值随体外培养时间的延长呈现由弱变强再变弱的变化规律,平均吸光度于第5天最高,第7,9天细胞周期素E呈下降趋势。结论:细胞周期素E表达量增加与人肾间质成纤维细胞的体外增殖速度有密切关系,可能在肾间质纤维化形成过程中发挥重要作用。更多还原

【Abstract】 AIM: To investigate the relationship between the expression of cyclin E and the proliferation of human renal interstitial fibroblasts (hRIFs) in vitro. METHODS: The experiment was conducted in the Laboratory of Pathology and Central Laboratory of Luzhou Medical College between June 2004 and October 2005. hRIFs of the third logarithmic phase were adopted for experiment. Serial cultivation was used to obtain and identify the hRIFs. Strept actividin-biotin complex (SABC) and image analysis were used to detect the total number of hRIFs and the number of hRIFs that positively expressed in cyclin E as well as the mean absorbance of positive cell respectively on the 1st, 3rd, 5th, 7th and 9th days after culture in vitro. Comparisons in the mean absorbance in cyclin E at each time-point between each two were done by q test. RESULTS: ① The total number of hRIFs increased gradually from the 1st to 9th day of cultivation in vitro, and cell growth curve was shown in an"S". The total number increased rapidly from the 3rd to the 5th day, which was the logarithmic phase, while the number from the 7th to the 9th day increased slowly. ② The ratio of hRIFs positively expressed in the cyclin E peaked on the 3rd-5th days, which was then relatively increased , while the positive ratio of hRIFs began to decreased slowly. ③ The mean absorbance in cyclin E was in a weaker-stronger-weaker changing rule with the time of cultivation prolonging. The mean absorbance peaked on the 5th day and decreased on the 7th and 9th days in cycline E. CONCLUSION: Increased expression of cyclinE is closely related with the proliferation of hRIFs in vitro, which may play an important role in the course of renal interstitial fibrosis.更多还原