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目的 :通过热休克预处理提高人牙髓细胞HSP70的表达水平 ,建立供体外实验研究的热休克细胞模型。方法 :将体外培养的人牙髓细胞爬片后进行热处理 ,置 42℃水浴 2 5min ,37℃复温。于复温后 1、2、3、4、6、8、10h、1d固定 ,进行HSP70的免疫组化染色 ;96孔板细胞同样方法热处理 ,连续培养 3d后 ,进行MTT检测。结果 :正常对照牙髓细胞呈阴性表达。热休克处理牙髓细胞 1、2h组中度阳性着色 ,3、4h组强阳性着色 ,6、8、10h组达到高峰 ,1d组恢复到弱阳性水平。MTT检测显示热处理组与对照组无统计学差异。结论 :热预处理方法可成功诱导人牙髓细胞表达HSP70 ;该方法不会导致连续培养 3d后细胞存活数量的明显变化
OBJECTIVE: To enhance the expression of HSP70 in human dental pulp cells by heat shock preconditioning and to establish a heat shock cell model for in vitro studies. Methods: Human dental pulp cells cultured in vitro were heat-treated and placed in 42 ℃ water bath for 25min and 37 ℃ rewarming. The cells were fixed at 1, 2, 3, 4, 6, 8, 10h and 1d after rewarming for immunohistochemical staining of HSP70. The 96-well plate cells were heat-treated in the same way and cultured for 3 days. MTT assay was performed. Results: The normal control dental pulp cells were negative expression. Heat shock treatment of dental pulp cells 1,2 h group moderate positive staining, 3,4h group strong positive staining, 6,8,10 h group peaked, 1d group returned to a weak positive level. MTT test showed no significant difference between the heat-treated group and the control group. CONCLUSION: Thermal preconditioning can successfully induce HSP70 expression in human dental pulp cells. This method does not result in a significant change in cell survival after 3 days of continuous culture