Objective: To observe the therapeutic effects and related mechanism of hemin on the progression of hepatic fibrosis in rats. Methods: Sixty male Wistar rats were randomly divided into normal control group, 4-week model group, 6-week model group, hemin inhibitor zinc protoporphyrin-IX (ZnPP-IX) intervention group and hemin intervention group. Hemin intervention group in complex liver fibrosis model was intraperitonealy administered ZnPP-IX or hemin every other day for 2 weeks from the fourth week. The mRNA expression of HO-1, α-smooth muscle actin (α-SMA) and nuclear factor-κB (NF-κB) in the liver tissue was detected by real-time polymerase chain reaction. Immunohistochemistry was used to detect HO-1 and localization of α-SMA expression. Serum hyaluronic acid, propeptide of type III collagen and hepatic transforming growth factor beta (TGFβ), and interleukin 6 (IL-6) expressions were detected by enzyme-linked immunosorbent assay. The content of hydroxyproline in hepatic tissues was measured by alkaline hydrolysis method. One-way ANOVA was used to compare the mean of each group. The difference between the two groups was compared by independent samples t- test. P-values < 0.05 was considered statistically significant. Results: Compared with model groups and ZnPP-IX intervention group, Hemin's intervention significantly increased the expression of HO-1 mRNA (P < 0.01) and protein distribution in liver tissues, while the expression of alpha-SMA mRNA was significantly decreased (P < 0.05) in portal space and areas around the fibrotic septum, and hepatic sinus. Hyp content and serum hyaluronic acid and propeptide of type III collagen decreased significantly (P < 0.05). Meanwhile, NF-κB p65 mRNA expression and the downstream production of TGFβ and IL-6 in Hemin intervention group were also inhibited (P < 0.05). Conclusion: Hemin can significantly inhibit the progression of hepatic fibrosis in rats by up-regulating HO-1 expression, and the inhibiting activity of NF-κB p65 leads to downstream of the inflammatory factors.
目的： 观察血晶素（Hemin）在大鼠肝纤维化进展过程中的治疗效果并探讨其相关作用机制。 方法： 将60只雄性Wistar大鼠随机分为正常对照组、4周模型组、6周模型组、血红素加氧酶-1（HO-1）抑制剂锌原卟啉-IX（ZnPP-IX）干预组、Hemin干预组，在复合因素构建肝纤维化模型过程中，干预组自第4周起开始给予隔日腹腔注射ZnPP-IX或Hemin干预2周。采用实时定量聚合酶链反应检测肝组织HO-1、α-平滑肌肌动蛋白（α-SMA）及核转录因子-κB（NF-κB）的mRNA表达；采用免疫组织化学技术检测HO-1及α-SMA定位表达；酶联免疫吸附法测定检测血清透明质酸、Ⅲ型胶原前肽及肝组织转化生长因子β（TGFβ）、白细胞介素6（IL-6）的表达；碱水解法检测肝组织羟脯氨酸（Hyp）含量。应用单因素方差分析（One-way ANOVA）进行各组样本均数的比较，两组间差异比较采用两独立样本的t检验，P < 0.05为差异有统计学意义。 结果： Hemin的干预同模型组及ZnPP-IX处理组相比可显著促进肝组织中HO-1的mRNA表达（P < 0.01）及蛋白分布，而α-SMA的mRNA表达（P < 0.05）及在汇管区、纤维间隔与肝窦周围分布明显减少，Hyp含量及血清中透明质酸、Ⅲ型胶原前肽水平均显著下降（P < 0.05）；同时Hemin干预组NF-κB p65的mRNA表达及其下游TGFβ、IL-6的产生也受到抑制（P < 0.05）。 结论： 血晶素可以显著抑制大鼠肝纤维化进展，该作用同其上调HO-1表达，进而抑制NF-κB p65的活性及下游炎性因子的释放密切相关。.
Keywords: Heme oxygenase-1; Hemin; Liver cirrhosis; Nuclear factor-κB.