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目的研究四氯化碳(CCl4)诱导性肝硬化大鼠肝组织内葡萄糖调节蛋白78(GRP78)表达的变化。方法将健康雄性Wistar大鼠随机分为对照组(n=5)和肝硬化组(n=5),对照组皮下注射植物油0.12ml/100g体重,肝硬化组以0.3ml/100g体重皮下注射60%CCl4植物油。在实验第21周的第1d杀死鼠,右心室取血检测肝功能指标。鼠肝中叶取材,石蜡切片行苏木精-伊红(HE)染色和天狼猩红组织化学染色,光镜观察形态学改变,行GRP78免疫组织化学显色,无菌下取肝组织行Western blotting及RT-PCR检测肝组织GRP78的表达。结果肝硬化组的血清ALT含量(278.2±88.42)明显高于对照组(154.8±9.94,t=3.10,P<0.05);ALB含量(1.68±0.62)明显低于对照组(3.02±1.96,t=2.62,P<0.05)。对照组肝小叶结构和肝细胞状态正常,仅在中央静脉及汇管区可见少量胶原纤维。20周肝硬化组的肝细胞脂肪变性明显,胶原纤维大量增生,假小叶形成。GRP78免疫组织化学呈色显示,肝硬化组肝组织中阳性细胞数明显多于对照组,棕黄色着色定位于细胞质中。Western blotting和RT-PCR显示肝硬化组肝组织GRP78的表达明显增强。结论在CCl4诱导性肝硬化的肝组织中,GRP78表达明显增强,这可能与肝细胞的内质网应激有关。
Objective To investigate the changes of glucose regulatory protein 78 (GRP78) expression in hepatic tissue of carbon tetrachloride (CCl4) -induced cirrhosis in rats. Methods Healthy male Wistar rats were randomly divided into control group (n = 5) and liver cirrhosis group (n = 5). The control group was subcutaneously injected with 0.12 ml / 100 g vegetable oil, and the liver cirrhosis group was subcutaneously injected with 0.3 ml / 100 g body weight % CCl4 vegetable oil. In the first 21 days of the experiment to kill rats, right ventricular blood test for liver function indicators. The middle part of the rat liver was harvested, and paraffin sections were stained with hematoxylin-eosin (HE) and Sirius red staining. Morphological changes were observed by light microscopy. GRP78 immunohistochemical staining was performed. The expression of GRP78 in liver tissue was detected by RT-PCR. Results The level of ALT in cirrhosis group was significantly higher than that in control group (278.2 ± 88.42, 154.8 ± 9.94, t = 3.10, P <0.05) = 2.62, P <0.05). The control group hepatic lobule structure and normal liver cells, only in the central vein and portal area visible a small amount of collagen fibers. 20 weeks of liver cirrhosis group of hepatocellular steatosis significantly, a large number of collagen fibers hyperplasia, pseudolobule formation. GRP78 immunohistochemical staining showed that liver cirrhosis group was significantly more than the number of positive cells in the control group, brown yellow staining located in the cytoplasm. Western blotting and RT-PCR showed that the expression of GRP78 in liver cirrhosis group was significantly increased. Conclusion The expression of GRP78 in CCl4-induced liver cirrhosis is significantly increased, which may be related to the endoplasmic reticulum stress in hepatocytes.