低强度周期性静水压力对人膝关节软骨细胞的影响  

Effects of Low-intensity Cyclical Hydrostatic Pressures on Chondrocytes of the Keen Joint in Human

作　　者：王鑫 白倩[2] 颜世举[3] 裘秀春[3] 范清宇[3] 马保安[3] 

机构地区：[1]兰州军区临潼疗养院第二疗区,陕西西安710600 [2]中国人民解放军第518医院,陕西西安710005 [3]第四军医大学唐都医院全军骨科中心暨全军骨肿瘤研究所,陕西西安710038

出　　处：《现代生物医学进展》2016年第21期4032-4036,4046,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81072194)

摘　　要：目的:探讨低强度周期性静水压力对体外培养的人膝关节软骨细胞增殖、凋亡,以及细胞Ⅱ型胶原分泌表达的影响。方法:体外酶消化法分离培养成人膝关节正常软骨细胞,将培养的第3代软骨细胞分为两组:正常对照组、3.0MPa组压力实验组,应用多功能恒温体外细胞培养中高压静水压力加载装置加载低强度周期性压力,共5d,每天2h。Ⅱ型胶原免疫组织化学染色法和甲苯胺蓝染色法鉴定软骨细胞,流式细胞术检测细胞凋亡,四甲基偶氮唑蓝(MTT)法绘制细胞生长曲线,qRT-PCR、Western-Blot检测Ⅱ型胶原的分泌和表达。结果:软骨细胞Ⅱ型胶原免疫组织化学染色和甲苯胺蓝染色均显示为阳性。与正常对照组相比,3.0MPa组表现出促进软骨细胞增殖,抑制细胞凋亡,且Ⅱ型胶原的合成分泌明显升高(P<0.05)。结论:通过体外模拟人生理情况下较低强度(3.0MPa)的周期性静水压力对人软骨细胞增殖、凋亡水平及周围基质分泌合成功能的影响,初步证实了较低强度压力有助于软骨自我修复和自身保护作用的发挥。Objective： To investigate the effects of low-intensity cyclical hydrostatic pressures on the proliferation, apoptosis and II collagen contents of chondrocytes of the keen joint cultured in vitro in human. Methods： The normal chondrocytes of human knee joint were isolated and cultured in vitro. The third generation of chondrocytes were treated with low-intensity cyclical hydrostatic pressures（3.0 MPa） by the multifunctional thermostatic high-insensitive hydrostatic pressure loading device for 2 h per day lasting for 5 days. Toluidine blue staining and immunohistochemical staining of type II collagen were employed to identify the chondrocytes. Cell apoptosis and proliferation were examined by flow cytometry and MTT assay respectively. The q RT-PCR and Western-Blot were performed to measure contents and expression of Ⅱ collagen of chondrocytes in two groups. Results： Immunohistochemistry and toluidine blue staining showed a positive staining of the chondrocytes. Compared with the control group, the 3.0 MPa group promoted proliferation, Ⅱ collagen contents and expression of chondrocytes, while inhibited apoptosis of chondrocytes（P〈0.05）. Conclusion： The lower-intensity cyclical hydrostatic pressure（3.0 MPa） under physiological conditions which can influence proliferation and apoptosis of human chondrocytes,stimulate the synthetic and secretory function in vitro experiments. Initially confirmed that the lower-intensity cyclical hydrostatic pressure significantly promote the Self-Healing and protective function of articular cartilage.

关 键 词：人软骨细胞 膝关节 低强度压力 Ⅱ型胶原 

分 类 号：R-33[医药卫生] R684.3