重型肝炎患者人工肝治疗前后血清对TGF-β1表达的影响

摘要

Objective To investigate the effect of plasma exchange to the expression of TGF-β1 in Kupffer cells ( KCs) . Methods A total of 40 patients with chronic severe hepatitis were treated with plasma exchange. The serums before and after treatment were collected. The level of serum lipopolysaccharide ( LPS) was measured by tachypleus amebo-cyte lysate method. The level of TGF-β1 was measured by enzyme linked immunosorbent assay ( ELISA) . KCs of rats were isolated and primarily cultured by liver perfusion with collagenase Ⅳ, density gradient centrifugation with Percoll and selective attachment. Peroxidase stain and phagocytosis of latex-beads test were used to identify KCs. The expression of TGF-β1 in KCs after co-cultured with serum for 24 hours was measured by immunohistochemical method. Results The levels of LPS and TGF-β1 in patients with chronic severe hepatitis were significantly higher than normal group ( P < 0. 01) , and were significantly decrease after plasma exchange treatment ( P < 0. 05 ) . Liver function and blood coagulation function obviously improved after plasma exchange treatment. The expression of TGF-β1 in KCs co-cultured with serum after plasma exchange treatment was significantly decreased ( P < 0.05). Conclusion Plasma exchange can not only directly decrease the level of TGF-β1, but also decrease the expression of TGF-β1 due to attenuate the activation of KCs.%目的 探讨血浆置换治疗前后慢性重型肝炎患者血清对大鼠枯否细胞(KCs)表达转化生长因子β1(TGF-β1)的影响.方法 对40例重型肝炎患者给予血浆置换治疗,收集治疗前后血清.基质偶氮显色法检测内毒素,酶联免疫吸附法(ELISA)检测TGF-β1浓度.Ⅳ型胶原酶灌注,梯度离心法分离大鼠KCs,用聚苯乙烯乳珠吞噬实验和过氧化物酶染色进行鉴定.治疗前后血清直接干预KCs,24 h后采用免疫细胞化学方法检测TGF-β1在KCs的表达.结果 慢性重型肝炎患者血清内毒素和TGF-β1浓度较健康者升高(P＜0.01)；血浆置换治疗后较治疗前降低(P＜0.05)；治疗后肝功能、凝血功能明显改善(P＜0.05)；治疗后患者血清刺激KCs产生TGF-β1较治疗前减少(P＜0.05).结论 血浆置换治疗可以通过直接清除作用以及减少内毒素对KCs的刺激,从而降低TGF-β1浓度,减少其对肝细胞再生的抑制作用,为肝细胞再生创造条件.