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目的通过系统测定大鼠肝硬变病程中血清透明质酸 HA 的动态变化,来探讨其用于肝纤维化的测试和监控的临床价值.方法利用 CCl_4损伤 SD 大鼠造成肝硬变模型(n=6),综合分析不同时间取材的实验组和正常对照组(n=6)之间血清肝纤维化指标-血清 HA 浓度、肝组织病理改变以及 HA 在肝组织中的免疫组织化学定位.结果在大鼠肝硬变的病变发展过程中,血清 HA 含量在3(7.98 ng/mL),6(20.10 ng/mL),9(229.73 ng/mL),12(324.74 ng/mL)周时,均较正常组(0周:0.21 ng/mL)有极显著升高(P<0.01),其中9 wk 和12 wk 的血清 HA 含量比正常对照组分别高出1094和1546倍.而在0→3 wk、3→6 wk、6→9wk、9→12wk 时,血清 HA 平均增加的倍数依次为38,2.5,11.4和1.4倍.同时,肝组织病理学结果(HE 染色)显示,CCl_4诱发的肝纤维化大鼠在9 wk 以前无明显变化;9 wk～12 wk 时,与正常肝组织相比,有明显肝细胞坏死,同时伴有肝细胞质分布不均匀程度的加深及红细胞增多成团现象.此外,12 wk 组还出现明显的肝细胞气球样变性现象及局部炎症细胞浸润.同期用免疫组化法观察 HA 在不同期肝硬变大鼠肝内的分布,镜检发现只在12 wk 有少量 HA 的特异性分布.结论动态观察血清 HA 数值可用于早期肝纤维化的预测和肝纤维化的分期.
Objective To investigate the clinical value of serum hyaluronic acid (HA) in the pathogenesis of liver cirrhosis by measuring the dynamic changes of serum hyaluronic acid (HA) in rat liver cirrhosis.Methods The CCl_4-injured SD rats were used to induce liver cirrhosis model (n = 6), the serum levels of liver fibrosis, the pathological changes of liver tissue and the immunohistochemical localization of HA in the liver tissue were analyzed between the experimental group and the normal control group (n = 6) During the course of the development of liver cirrhosis in rats, the serum levels of HA were significantly higher at 3 (7.98 ng / mL), 6 (20.10 ng / mL), 9 (229.73 ng / mL) and 12 (324.74 ng / (P <0.01). The contents of HA in serum of 9 wk and 12 wk were 1094 and 1546 times higher than that of normal control group respectively The mean increase of serum HA was 38, 2.5, 11.4 and 1.4 times in the order of 3 wk, 3 → 6 wk, 6 → 9 wk and 9 → 12 wk, respectively.Hepatic histopathological results (HE staining) showed that CCl 4 -induced Compared with normal liver tissue, hepatic fibrosis rat had no obvious change before 9 wk. At 9 wk ~ 12 wk, compared with normal liver tissue, there was obvious hepatocyte necrosis accompanied with hepatocyte cytoplasmic distribution Homogeneity deepening and erythrocytosis.In addition, 12 wk group also showed obvious phenomenon of hepatocyte balloon-like degeneration and local inflammatory cell infiltration.At the same period, immunohistochemistry was used to observe the effect of HA on the liver in different stages of liver cirrhosis rats , The microscopic examination showed that there was only a small amount of HA specific distribution at 12 weeks.Conclusions The dynamic observation of serum HA value can be used to predict the early stage of liver fibrosis and the stage of liver fibrosis.