常用柠檬酸钠抗凝血浆对HepG2细胞的损伤观察  

The injuries of sodium citrate plasma on HepG2 cells

作　　者：程永波[1] 王英杰[1] 张世昌[1] 陈志[1] 陈国致[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《中国医师杂志》2006年第5期605-607,共3页Journal of Chinese Physician

基　　金：国家自然科学基金资助项目(30470458)

摘　　要：目的观察常用柠檬酸钠抗凝血浆(scP)对HepG2细胞的损伤,为大量输注或置换血浆治疗晚期重型肝炎病人研究提供实验依据。方法复苏培养HepG2细胞。四甲基偶氮唑盐法测细胞活力,流式细胞仪测定细胞周期和凋亡率,检测细胞LDH漏出量和总蛋白、谷胱甘肽含量,观察培养过程中细胞形态变化,系统了解scP对体外培养HepG2细胞生长、损伤、功能和形态方面的影响。结果⑴各浓度scP培养第1d HepG2细胞活力下降(F=37.108,P=0.001);第2天除10%scP组外,50%、100%scP组细胞绝大部分死亡。⑵细胞生长周期测定50%scP培养1 d HepG2细胞S期所占比例较对照组增高,凋亡率10%scP和50%scP组都远高于对照组。⑶scP作用HepG2细胞5 h,LDH漏出量都高于RPM I-1640对照组(P值均<0.05),这种差异随浓度升高加大。⑷scP培养HepG2细胞24 h总蛋白合成下降,GSH含量升高。⑸光镜下100%scP培养24 h,HepG2细胞大部分死亡,10%scP组生长相对较好;100%hP组与对照组比较差异无统计学意义。结论scP对体外培养HepG2细胞有毒性作用,原因主要与血液保养液中所含的柠檬酸和柠檬酸钠有关。Objective To observe the injuries of sodium citrate plasma （scP） on the growth and function of HepG2 cells. Methods The HepG2 cells were cultured. The viability, cell cycle and apoptosis, the leakage of LDH, total protein, glutathione and the changes on morphology of hepatocytes exposured to scP were investigated. Results The viability of HepG2 cells was inhibited when the cells were cul- tured in scP for 24h （ F = 37. 108, P = 0. 001 ）. After 48h, nearly all cells died except cells in 10% scP group. After the cells were exposed to scP for 24h, the percentage of S phase of the cell cycle and the rate of apoptosis were significantly increased compared to those of the control. The leakages of LDH were increased in the HepG2 following exposure to scP for 5h （ P 〈0. 05）. Exposure of HepG2 cells to scP within 24h resulted in a decrease in the total protein synthesis and a increase in the GSH content. Conclusion scP is damaged to the growth and function of the HepG2 cells, which results from the citric acid and sodium citrate contained in anticoagulant.

关 键 词：柠檬酸盐类 抗凝药 肝细胞 肝炎 

分 类 号：R457.1[医药卫生—治疗学]