细胞表达HSP70的细胞模型研究

The study on expression of heat shock protein 70 in human dental pulp cell model through thermal preconditioning

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

AIM:To establish a heat shock cell model of human dental pulp cell, which containing promoted HSP70 by heat perconditioning. METHODS:The human dental pulp cells seeded on cover slip were preconditioned at 42℃ for 25min,and recovered at 37℃for 1,2,3,4,6,8,10h,and 1d respectively.The expression of HSP70 was studied by immunohistochemistry. The cells seeded in 96-well plates were preconditioned using the same method,and then cultured 3 days for MTT assay.RESULTS:The control group was negative.The 1h and 2h groups were moderate positive. The 3h and 4h groups were strong positive. The immunohistochemistric staining reached the strongest at 6,8h and 10h . After 24h the expression of HSP70 droped back to less positive. There was no statistical difference between preconditioning group and control group in MTT assay. CONCLUSION: The preconditioning method can successfully induce the expression of HSP70 in human dental pulp cells. The thermal preconditioning will not lead to change in number of survival cells cultured for 3 days.