Hsp27mrna, Hsp70mrna Expression In Cultured Human Lens Epithelial Cells

Objective:Investigation of the pathogenesis of cataract through the observation of the effect by high-temprature oxidation on the expression of heat shock protein in the human lens epithelial cells,try to prove that HSP27,HSP70 could participate in the process of the prevention of crystalline lens.Methods:Human HLB-C3's cell line(from Chinese medicine department university subsidiary the ophthalmology department of the first hospital provide) contained 15% calf serums was cultivated within the DMEM 37℃culture box .Then the lens cell was divided into the control group and two experimental groups (Ⅰ,Ⅱ), Experimental groupⅠwas cultivated in 30 minutes with the high temperature (45℃) condition . In difference compartments (0 h, 2 h, 4 h, 6 h, 16 h, 24 h) we adopt the method of Reverse transcription polymerase chain reaction (RT-PCR) to examine HSP27mRNA, the HSP70mRNA level. Experimental groupⅡwas nurtured in oxidation (50mmol/LH2O2) condition in 30 minutes.After restoration to normal condition , in difference compartment (0h,2h,4h,6h,16h,24h) in the biological medium condition . we adopt the method of Reverse transcription polymerase chain reaction (RT-PCR) to examine HSP27mRNA, the HSP70 mRNA level. Comparison group in difference compartment (0h,2h,4h,6h,16h,24h) was adopted by the method of Reverse transcription polymerase chain reaction (RT-PCR) to examine HSP27mRNA, the HSP70mRNA level. Regardβ-actin as the confidential reference items to correct, and carry on statistics analysis with the index obtained.Results:Under the physiological,the high temperature and the state of oxidation condition the hot shock protein(HSP) can express. With the High temperature and the oxidized stress 2 hours latter they cause the increase of HSPmRNA level . The statistical analysis demonstrated that the expression of the HSP27,HSP70 in high temperature group and oxidation group are obviously higher than comparison group, it has significance difference (p<0.05). And in each group the HSP27 and HSP70 expression reached the peak value in different time-gap from 2h to 6h. .Later it would reduce gradually. But all of them still maintained high level in 16h. Conclusion:HSPs exsit in human lens epithelial cells and can be induced after stress. the data shows that it could pretect human lens epithelial cells in high-temprature oxidation status .