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生理周期性牵张应力促进人类膀胱平滑肌细胞增殖分子机制的研究

发表时间:2016-03-15  浏览量:1885  下载量:779
全部作者: 田野,岳轩,杨童欣,陈林,罗德毅,王坤杰
作者单位: 四川大学华西医院泌尿外科;成都大学附属医院泌尿外科
摘 要: 目的:探索不同模式的牵张应力条件促进人类膀胱平滑肌细胞(human bladder smooth muscle cells,HBSMCs)增殖能力的生物学效应,以及这一过程背后的分子生物学机制。方法:将HBSMCs接种到硅胶膜上,对其施加单轴拉伸应力,其应变参数分别为2.5%,5%,10%和15%,牵张频率参数分别为0.05,0.10,0.20,0.50,1.00 Hz,并设置无应力施加的对照组进行研究。采用溴-脱氧尿嘧啶(bromodeoxyuridine,BrdU)掺入法量化分析细胞的增殖能力,摸索最适合HBSMCs增殖的牵张模式。采用实时PCR(real-time PCR,RT-PCR)技术和Western blotting技术分别从分子水平和蛋白质水平检测生理周期性牵张应力条件下HBSMCs的PI3K、SGK1、Akt、Kv1.3分子表达情况,以进一步研究应力促进细胞增殖的分子机制。结果:最适合HBSMCs增殖的牵张应力参数为应变5%、牵张频率0.10 Hz,因该试验组的细胞增殖参数较其他各试验组和对照组更高,其结果具有统计学意义(P<0.05)。与对照组相比,牵张试验组的PI3K、SGK1、Kv1.3表达量显著升高(P<0.05),而Akt则无明显改变(P>0.05)。分别应用SGK1和Kv1.3的阻滞剂重复细胞牵张试验均可消除上调的增殖能力。结论:周期性牵张应力可以促进HBSMCs增殖,其力学分子生物学信号可能通过PI3K-SGK1-Kv1.3通路转导,而PI3K-Akt通路并未参与其中。
关 键 词: 泌尿外科学;组织工程;平滑肌细胞;细胞增殖;机械应力;信号转导
Title: Physiological cyclic stretch induced proliferation of human bladder smooth muscle cells via PI3K-SGK1-Kv1.3 pathway
Author: TIAN Ye, YUE Xuan, YANG Tongxin, CHEN Lin, LUO Deyi, WANG Kunjie
Organization: Department of Urology, West China Hospital, Sichuan University; Department of Urology, Affiliated Hospital of Chengdu University
Abstract: Objective: To explore the different modes of stress induced proliferation of human bladder smooth muscle cells (HBSMCs) and to investigate the mechanism involved in this process. Methods: HBSMCs were seeded in a silicone membrane and subjected to cyclic stretch of 2.5%, 5%, 10% and 15% equibiaxial strain at frequencies of 0.05, 0.10, 0.20, 0.50, 1.00 Hz, respectively. HBSMCs without stess were employed as the control group. Bromodeoxyuridine (BrdU) assays were used to detect the proliferative activity of each group, and hereby to determine the optimal stress mode. To further determine the mechanism of the cell proliferation process triggered by physiological cyclic stretch, the expression of PI3K, SGK1, Akt, Kv1.3 was investigated at the transcriptional and translational levels (molecular and protein levels) by real-time PCR (RT-PCR) and Western blotting analysis, respectively. Results: Optimal physiological stretch was established as 5% strain at a frequency of 0.10 Hz, whereby HBSMCs revealed a statistical significant increase in proliferative activity compared with the other groups, including the control group, which served as the control (P<0.05). The expressions of PI3K, SGK1 and Kv1.3 (P<0.05), however, without Akt (P<0.05), were upregulated by cyclic stretch as compared with the control group. When separately treated with inhibitors of SGK1 and Kv1.3, increased stress induced proliferation was largely eliminated. Conclusion: Cyclic stretch induces the proliferation of HBSMCs and the PI3K-SGK1-Kv1.3 pathway is involved in this process, either fully or at least partially, rather than its related pathway, PI3K-Akt.
Key words: urology; tissue engineering; smooth muscle cells; cell proliferation; mechanical stress; signal transduction
发表期数: 2016年3月第5期
引用格式: 田野,岳轩,杨童欣,等. 生理周期性牵张应力促进人类膀胱平滑肌细胞增殖分子机制的研究[J]. 中国科技论文在线精品论文,2016,9(5):428-434.
 
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2016-04-06 14:12:41
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