Kupffer细胞NF-κB激活对胆道梗阻合并肝部分切除鼠肝细胞再生的影响

Effects of Kupffer cell NF-κB activation on liver regeneration after partial hepatectomy in biliary obstructive rats

目的 探讨Kupffer细胞 (KCs)NF κB激活在鼠非肝硬变性胆道梗阻肝部分切除后肝细胞再生过程中的作用。方法 Wistar大鼠随机分为正常大鼠 70 %肝部分切除组 (N PH)、胆总管梗阻 1周 70 %肝部分切除组 (BDO PH)、BDO PHIL 1 0或生理盐水治疗组。EMSA法检测KCsNF κB激活、RT PCR法检测肝内HGF、IL 6、TGF β1及IL 1 βmRNA表达 ,ELISA法检测肝内TNF α水平 ,免疫组化法检测肝细胞BrdU标记。结果 BDO PH后 0～ 72h肝内IL 6mRNA、TGF β1mRNA、IL 1 βmRNA及TNF α表达增高而HGFmRNA表达减少 ,肝细胞BrdU标记减少。而IL 1 0能下调BDO PH后KCsNF κB活性、上调肝内HGFmRNA与下调肝内IL 6mRNA、TGF β1mRNA、IL 1 βmRNA及TNF α表达 ,从而促进肝细胞BrdU标记。结论 KCsNF κB过度激活可能抑制非肝硬变性胆道梗阻肝部分切除后肝细胞再生 ,适当调节KCsNF κB活性可能促进肝细胞再生。

Objective To explore the effects of Kupffer cells(KCs) NF κB activation on liver regeneration after 70% partial hepatectomy(PH) in rats with noncirrhotic bile duct obstruction(BDO). Methods Wistar rats were randomly divided into normal rats 70%PH group (N PH), BDO PH group in which 70%PH were performed after common bile duct obstruction for 1 week, BDO PH IL 10 treatment group and sham control group in which recombinant murine IL 10 (10 mg/kg) or normal saline were injected via the dorsum vein of penis per 24 h after BDO. NF κB activation of KCs were determined with electrophoretic mobility shift assay (EMSA). Expression levels of hepatic HGF(Hepatocyte growth factor) mRNA, IL 6 mRNA, IL 1β mRNA,TGF β 1 mRNA and TNF α were measured with RT PCR and ELISA. BrdU labelling of hepatocytes were determined with immunohistochemical analysis. Results NF κB activities of KCs , expression levels of hepatic IL 6 mRNA, IL 1β mRNA, TGF β 1 mRNA and TNF α were significantly higher and the expression levels of hepatic HGF mRNA were significantly lower in BDO PH group than in N PH group rats from 0～72 h after 70%PH. BrdU labelling of hepatocytes reached the peak at 24 h after 70%PH in N PH group, and at 48 h in BDO PH group, and the peak levels of BrdU labelling of hepatocytes in BDO PH group were decreased significantly compared with those of N PH group; NF κB activities of KCs , expression levels of hepatic IL 6 mRNA, IL 1β mRNA, TGF β 1 mRNA and TNF α were significantly lower and the expression levels of hepatic HGF mRNA were significantly higher in BDO PH IL 10 treatment group rats than in BDO PH group rats from 0～72 h afte 70%PH. BrdU labelling of hepatocytes reached the peak at 48 h after 70%PH in BDO PH IL 10 treatment group, and the peak levels of BrdU labelling of hepatocytes in BDO PH IL 10 treatment group were increased significantly compared with those of BDO PH group. Conclusion The excessive NF κB activation of KCs after BDO PH inhibited regeneration of hepatocytes , and adjustment of KCs NF κB activities to a appropriate level could promote liver regeneration after BDO PH.