体外灌流慢性重型肝炎患者血浆对C3A细胞的细胞色素P4503A活性和表达的影响

目的：通过比较未经灌流处理的重肝血浆及处理过的重肝血浆对CYP3A的影响，阐明C3A细胞安定代谢变化的原因，为未来C3A细胞的改造与功能优化奠定基础。方法：制备血浆和培养C3A细胞；实验分为4组：正常胎牛血浆（NFBP）组，正常人血浆（NHP）组，体外灌流慢性重型肝炎患者血浆（HPP）组，体外未灌流慢性重型肝炎患者血浆（CSHP）组。用分光光度法测定红霉素N-脱甲基酶（ERD）活性即CYP4503A的活性，Western blot法测定CYP4503A4的表达。结果：CSHP组ERD活性低于NFBP组（P〈0．05）及NHP组（P〈0．05），HPP组ERD活性也低于NFBP组（P〈0．05）及NHP组（P〈0．05），HPP组ERD活性高于CSHP组（P〈0．05）；CSHP组CYP4503A4的表达低于NFBP组（P〈0．05）及NHP组（P〈0．05），HPP组CYP4503A4的表达也低于NFBP组（P〈0．05）及NHP组（P〈0．05），HPP组CYP4503A4的表达高于CSHP组（P〈0．05）。结论：经慢性重型肝炎患者血浆培养的C3A细胞，CYP4503A活性及蛋白表达降低。与之相比，经过体外灌流的慢性重型肝炎患者血浆其C3A细胞CYP4503A活性及蛋白表达有所增加，CYP4503A活性增加可能是导致安定代谢变化的原因。

Effect of hemoperfusion treated plasma from patients with chronic severe hepatitis on the activity and expression of CYP4503A in C3A cells

AIM: To investigate the changes of CYP4503A, a key enzyme of diazepam metabolism, and explore the effect of hemoperfusion treated plasma from patients with chronic severe hepatitis on the activity and expression of CYP4503A in C3A cells. METHODS: Plasma was prepared and C3A cells were cultured. There were four groups in the experiment: normal fetal bovine plasma (NFBP) group, normal human plasma (NHP) group, hemoperfusion plasma (HPP) group, chronic severe hepatitis plasma (CSHP) group. The activity of erythromycin N-demethylase (ERD), namely the activity of CYP4503A, was measured by spectrophotometer and the expression of CYP4503A4 was detected by Western blot. RESULTS: The activity of ERD in CSHP group and HPP group was lower than that in NFBP group (P<0.05) and NHP group (P<0.05) but the activity of ERD in HPP group was higher than that in CSHP group (P<0.05). The expression of CYP4503A4 increased in NFBP group (P<0.05) and NHP group (P<0.05) but the expression of CYP4503A4 in CSHP group decreased compared with HPP group(P<0.05). CONCLUSION: The activity of ERD and the expression of CYP4503A decreased in CSHP group while the activity of ERD and the expression of CYP4503A in HPP group increased, which may cause the changes of the diazepam metabolic rates of C3A cells. This finding will provide some references for future researches on cell construction.