›› 2004, Vol. 3 ›› Issue (9): 496-500.

• 人工肝 • 上一篇    下一篇

重型肝炎患者血浆对HepG2细胞的损伤作用

程永波 张世昌 王英杰 陈 志 刘 俊 李嘉佳   

  1. 400038 重庆,第三军医大学附属西南医院全军感染病研究所
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2004-09-12 发布日期:2004-09-12

  • Received:1900-01-01 Revised:1900-01-01 Online:2004-09-12 Published:2004-09-12

摘要:

目的 观察重型肝炎患者血浆( severe hepatitis plasma,SHP)对HepG2细胞的损伤,为保持生物人工 肝反应器内肝细胞材料活力研究提供实验依据,以期提高生物人工肝的临床治疗效果。方法 复苏培养HepG2细胞,通过MTT法测细胞活力、检测LDH漏出量和细胞凋亡率及观察形态变化四个方面了解SHP对HepG2细胞的损伤情况。结果 ①所有浓度SHP培养的HepG2细胞活力都下降,100%SHP组下降幅度最大;②HepG2细胞在SHP作用下乳酸脱氢酶(LDH)漏出量与对照组相比有显著性差异,并且差异随时间延长加大;③不管灭活与否、浓度高低,SHP培养的HepG2细胞凋亡率都远高于对照组,最高为灭活前100%SHP组;④SHP可致HepG2细胞形成膜大疱,高浓度时致较多细胞死亡。结论 SHP能够损伤HepG2细胞,在生物人工肝的临床应用中,有必要对反应器中的肝细胞采取防护措施。

关键词: HepG2 细胞, 细胞培养, 重型肝炎, 血浆, 损伤

Abstract:

Objective To observe the injuries of plasma from patients with severe hepatitis on HepG2 cells. For the advanced study on hepatocytes to maintain their function in a bioartificial liver device and improving the clinical efficiency of the bioartificial liver device. Methods HepG2 cells were cultured. Viability, the leakages of LDH, apoptosis and the changes on morphology of HepG2 cells exposured to SHP were measured and investigated. Results ①The viability was inhibited in 10%, 30%, 50%, 100% SHP, a significant difference occurred in the culture medium containing 100% SHP compared to the controls. ②the leakages of LDH was significantly increased in the HepG2 cells cultured with SHP, at the end of the experiment they were approximately 2 times higher than that of HepG2 cells cultured in the medium containing 10% NCS. ③Exposure to SHP,especially the 100% SHP, resulted in a signifiant increase in apoptosis of HepG2 cells compared to control cultures. ④The bullae were observed from SEM pictures of HepG2 cells cultured in SHP for 24h. The higher of the SHP concentration the more dead cells were found. Conclusion Plasma from patients with severe hepatitis can damage HepG2 cells. It could be necessary to pretreat the patients,plasma to increase the efficiency of the bioartificial liver device. 

Key words: Cell culture, Severe hepatitis, Plasma, Injury

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