脾脏在大鼠肝硬化形成过程中免疫调控机制的探讨

The mechanism for splenic promoting effects on liver cirrhosis

为了解脾脏促进肝硬化形成的机理，作者用ＣＣｌ４诱导大鼠肝硬化模型，实验动物切脾或注促吞噬肽（Ｔｕｆｔｓｉｎ），同时分离肝实质、枯否（Ｋｕｐｆｆｅｒ）和贮脂（Ｉｔｏ）细胞，加ＣＣｌ４和脾条件液或Ｔｕｆｔｓｉｎ混合培养；并由动物模型肝组织块提取ＲＮＡ，同５种ｃＤＮＡ探针作分子杂交，从整体、细胞和分子三个不同水平探讨其作用机制。结果表明，在肝硬化模型，与切脾组相比，有脾组动物血中白细胞介素１（ＩＬ１）、ＩＬ６和肿瘤坏死因子（ＴＮＦα）升高（Ｐ＜０．０５）。体外Ｉｔｏ细胞培养发现，加入脾条件液或Ｔｕｆｔｓｉｎ比单独加ＣＣｌ４的细胞分泌较多的纤维连接蛋白、层粘连蛋白和Ｉ型胶原（Ｐ＜０．０５）。狭缝杂交结果表明，从有脾组动物提取的ＲＮＡ与ＴＮＦα、ＩＬ１β、转化生长因子（ＴＧＦ）β和ＣｏｌｌａｇｅｎＩ探针杂交比切脾组呈现更高密度的放射自显影图像。作者认为，脾脏促进肝硬化形成的免疫调控是一复杂的过程，促进ＴＧＦβ基因的表达可能是引起肝脏纤维过度增生的关键所在。

The model of liver cirrhosis was induced by CCl4 and alcohol in rats, which were subjected to splenectomy or given Tuftsin.The isolated and purified liver parenchymal, Kupffer and Ito cells were cultured with CCl4 and splenic conditional fluid or Tuftsin. The RNA isolated from the liver tissues of cirrhosis animals were hybridized with five kinds of cDNA probes. In this study we explored the mechanism of spleen's promoting effects on the liver cirrhosis formation at the whole body, cellular and molecular levels. The result showed that in cirrhosis model,the levels of IL1, IL6 and TNFαin serum of rats in imitative splenectomy or Tuftsin group were significantly increased compared to those in splenectomy group (P<0. 05 ). Cell culture showed that if medium contained CCl4 and splenic conditional fluid or Tuftsin, its cells can secrete more fibronectin,laminin and collagen I than those cultured in medium only contained CCl4 (P<0. 05). Slot blot hybridization showed that the RNA isolated from liver of rats in imitative splenectomy or Tuftsin group hybridized with probes of TNFα , IL1β, TGFβ and Collagen I had a more high density picture of X-ray than that isolated from liver of rats in the splenectomy group. Should be it a complex process for spleen to promote liver cirrhosis formation, in which the TGFβ gene expression enhancement may be the key of splenic effects on liver fibrosis.