苯巴比妥钠对环磷酰胺致大鼠肝细胞毒性的影响

Influence of phenobarbital on cyclophosphamide induced toxicity of rat liver cells

目的:利用原代培养正常大鼠肝细胞研究苯巴比妥钠(PNB)对环磷酰胺(CP)致离体大鼠肝细胞的毒性作用。方法:用两步灌注法分离原代大鼠肝细胞;观察苯巴比妥钠(PNB)对环磷酰胺(CP)致离体大鼠肝细胞的毒性作用。采用四甲基噻唑蓝(MTT)法测定PNB对原代大鼠肝细胞的最大无毒浓度(TC0),先用PNB的最大无毒浓度(TC0)诱导原代大鼠肝细胞24h,再直接加含不同浓度CP的培养液培养24h,MTT法测药物对细胞生长抑制率的影响,计算IC50。并测定药物作用后培养液上清中AST、ALT和LDH活性及培养液中GSH和细胞内GSH的含量。结果:经苯巴比妥钠诱导后的环磷酰胺各剂量组与未经苯巴比妥钠诱导的环磷酰胺各剂量组比较,细胞生长抑制率降低,培养液上清AST、ALT和LDH活性降低,培养液上清和细胞内GSH含量升高。结论:苯巴比妥钠能减轻环磷酰胺致离体大鼠肝细胞的毒性作用。

OBJECTIVE By using primary cell culture of normal adult rat hepatocytes,to study the toxicity of phenobarbital sodium （PNB） against cyclophosphamide in isolated rat liver cells. METHODS Using isolate hepatocytes of rat by two-step perfusion, the toxicity of phenbarbital sodium （PNB） against cyclophosphamide （CP） was observed in isolated rat liver cells. The largest non-toxic concentration TG, of the PNB on primary rat hepatocytes was determined by using MTT method. First, the largest non-toxic concentration of PNB was used to induce primary rat hepatocytes for 24 h,then different concentrations of CP was added into culture for 24 h. The inhibition of cell growth rate was determined by MTT,the IC50 was calculated and the activity of AST, ALT and LDH and the contents of GSH and GSH in cells culture were determined. RESULTS Compared all groups that have been incubated by the sodium phenobarbital with those groups without phenobarbital incubation, the cell growth inhibitory rate degraded, AST, ALT and LDH activity in the culture medium supernatant degraded, while the cell GSH content in culture medium supernatant increased. CONCLUSION Phenobarbital sodium can reduce toxicity in the cyclophos-phamide-induced rat liver ceils in vitro.