慢性重型肝炎患者血浆对HEPG_2细胞增殖及解毒功能的影响

Effects of plasma from patients with chronic severe hepatitis on the proliferation and detoxic capability of HEPG_2 cell line

目的:观察慢性重型肝炎患者血浆(CSHP)对HEPG2细胞增殖及解毒功能的影响。方法:分别用100%正常人血浆(NHP)和100%CSHP培养HEPG2细胞。培养24、48、72、96和120 h,MTT法检测细胞活力;培养24、48 h,姬姆萨染色,倒置显微镜下观察2组HEPG2细胞的形态;培养48 h,扫描电镜观察2组细胞的生长状态,并检测HEPG2细胞内还原型谷胱甘肽(GSH)含量。结果:连续5 d,100%CSHP培养的HEPG2细胞活力均高于100%NHP组(t=5.424、4.714、3.834、3.781和6.106,P均<0.001)。培养24、48 h,可见100%CSHP培养的HEPG2细胞生长较密。培养48 h,电镜下可见2组细胞生长状态均良好,但100%CSHP组处于分裂前期的细胞数多于100%NHP组,且分裂较活跃;100%CSHP组HEPG2细胞内还原型GSH含量为(306.7±16.2)mg/g,低于100%NHP组的(492.3±15.7)mg/g(t=9.18,P<0.001)。结论:CSHP可促进HEPG2细胞的增殖,并使HEPG2细胞内还原型GSH消耗明显增加,导致其解毒功能下降。

Aim： To investigate the effects of plasma from patients with chronic severe hepatitis on HEPG2 cell in vitro. Methods：HEPG2 cells were cuhured in normal human plasma（NHP） and chronic severe hepatitis plasma（CSHP） , respectively. MTT method was used to detect the activity of HEPG2 cell cultured for 24,48,72,96 and 120 h. Cellular mor- phology was observed by inverted microscope and electron microscope. The content of glutathione （GSH） inside the cells was detected after being cultured for 48 h. ReSUlts ：The activities of HEPG2 cells cultured in CSHP were higher than those in NHP for successive 5 days（ t = 5. 424,4. 714,3. 834,3. 781 , and 6. 106, P 〈 0. 001 ）. HEPG2 cells cultured in CHSP for 24 h looked thicker than that in NHP. The observation through electron microscope demonstrated that the HEPG2 cells cultured for 48 h in both groups grew well,while the ceils in prophase of division in CSHP group were much more than that in NHP group. The content of GSH inside HEPG2 cell cultured in CSHP for 48 h was （306.7 ± 16.2） rag/g, lower than that of （492.3± 15.7 ） mg/g in NHP （t = 9.18 ,P 〈 0. 001 ）. Gonclusion ： CHSP can not only promote the proliferation of HEPG2 cells, but also decrease the detoxic capability of HEPG2 cells by exhausting GSH inside the cells.