上海交通大学学报（医学版）

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

• 论著（基础研究） • 上一篇    下一篇

不同血液透析液对U937细胞凋亡和PKCδ表达的影响

朱小娟1, 郭黎莉2, 潘 瑜1, 谭龙益3, 张必嘏2, 金 燕2, 黄明珠2, 张国兰2, 蒙海燕2, 薄卫亚2, 徐琴君2, 金惠敏1   

  1. 上海交通大学 1. 医学院第三人民医院肾脏内科, 上海 201900；2. 第一人民医院宝山分院血液透析室, 上海 200940；3. 第一人民医院宝山分院检验科, 上海 200940
  • 出版日期:2009-12-25 发布日期:2009-12-25
  • 通讯作者: 郭黎莉, 电子信箱: gflmei@sohu.com。
  • 作者简介:朱小娟（1982—）, 女, 硕士生；电子信箱: zhuxiaojuan200709@126.com。
  • 基金资助:

    上海市卫生局基金（2007129）和上海市宝山区科委基金（08-E-28）

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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摘要：

目的 研究不同血液透析液对U937细胞蛋白激酶C-δ（PKCδ）表达及细胞凋亡的影响。方法 在对数生长期人单核细胞株U937细胞培养液中添加不同血液透析液进行干预，分为A液＋B液组（血液透析液A液＋B液组合）、A液＋B液+PKCδ特异性抑制剂（rottlerin）组、A液＋B粉组（血液透析液A液＋B粉组合）、A液＋B粉+ rottlerin组，同时设立空白对照和正常对照组。分组干预后24 h和48 h分别收集细胞，Hoechst33258染色后荧光显微镜观察细胞形态学改变；Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡；RT-PCR和Western blotting 检测PKCδ mRNA和蛋白表达。结果 A液+B粉组凋亡细胞明显增多，呈凋亡典型形态学改变，分组干预24 h和48 h的细胞凋亡率分别为（34.99±3.54）%和（46.35±4.22）%，明显高于相应时间点空白对照组的（7.40±1.33）%和（7.62±0.95）%、正常对照组的（5.37±0.29）%和（6.46±1.02）%和A液＋B粉+rottlerin组的（23.58±4.12）%和（33.70±4.50）%（P<0.05）；同时与两对照组和A液＋B粉+rottlerin组比较，A液＋B粉组U937细胞PKCδ mRNA和蛋白表达均明显上调（P<0.05）。A液＋B液组细胞凋亡率及PKCδ mRNA和蛋白表达与两对照组和A液＋B液+rottlerin组比较，差异均无统计学意义（P＞0.05）。结论 血液透析液A液＋B粉组合对U937细胞有明显促凋亡作用，其机制可能与上调细胞PKCδ表达有关；与A液＋B粉组合比较，选择A液＋B液组合作为血液透析液可降低患者外周血单核细胞的细胞凋亡率。

关键词: 透析液, U937细胞, 细胞凋亡, 蛋白激酶C-δ

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