›› 2009, Vol. 29 ›› Issue (12): 1434-.

• Original article (Basic research) • Previous Articles     Next Articles

Effects of different dialysates on apoptosis and expression of PKCdelta of U937 cell line

ZHU Xiao-juan1, GUO Li-li2, PAN Yu1, TAN Long-yi2, ZHANG Bi-gu2, JIN Yan2, HUANG Ming-zhu2, ZHANG Guo-lan2, MENG Hai-yan2, BO Wei-ya2, XU Qin-jun2, JIN Hui-min1   

  1. 1. Department of Nephrology, The Third People's Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 201900, China；2. Hemodialysis Unit, 3. Clinical Laboratory, Baoshan Branch, The First People's Hospital, Shanghai Jiaotong University, Shanghai 200940, China
  • Online:2009-12-25 Published:2009-12-25
  • Supported by:

    Shanghai Municipal Health Bureau Foundation, 2007129; Foundation from Science and Technology Committee of Baoshan District, Shanghai, 08-E-28

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Abstract:

Objective To investigate the effects of different dialysates on expression of protein kinase C-δ (PKCδ) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKCδ specific inhibitor）group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKCδ mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were (34.99±3.54)% and (46.35±4.22)%, respectively, significantly higher than those at corresponding time points in blank control group [(7.40±1.33)% and (7.62±0.95)%], normal control group [(5.37±0.29)% and (6.46±1.02)%] and fluid A+powder B+rottlerin group [(23.58±4.12)% and (33.70±4.50)%] (P<0.05). Compared with normal control group, blank control group and fluid A+powder B+rottlerin group, the expression of PKCδ mRNA and protein of U937 cells in fluid A+powder B group were significantly increased (P<0.05). There was no significant difference in cell apoptosis rates and expression of PKCδ mRNA and protein between fluid A+fluid B group and blank control group, normal control group and fluid A+fluid B+rottlerin group (P＞0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKCδ. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing  apoptosis of peripheral blood monouclear cells.

Key words: dialysate, U937 cell, cell apoptosis, protein kinase C-δ