摘要
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 research the expression of glucose regulating protein 78 （GRP78） in livers of carbon tetrachloride （CCl4） induced hepatoeirrhosis rats. Methods Healthy adult male Wistar rats were randomly divided into control group （n = 5） and hepatoeirrhosis group （n = 5）. Oily solution with 0.12ml per 100g rat weight was subcutaneous injected in control group, and 60% carbon tetrachloride oily solution with 0.3ml per 100g rat weight were subcutaneous injected in hepatoeirrhosis group for 20 weeks. At the first day of 21 weeks, all rats were killed. Each blood was collected from right ventricle and some hepatic function was measured. The morphological structure in hepatic middle lobule were observed through the staining of H E and Sirius red. The expressions of GRP78 was detected by immnunohistoehemistry, Western blotting and RT-PCR. Results The serum ALT in 20 weeks hepatoeirrhosis group （278.2±88.42） was evidently higher than that in control group （154.8±9.94, t = 3.10, P 〈 0.05）, and the serum ALB in the 20 weeks hepatoeirrhosis group （1.68±0.62） was evidently lower than that of control group （3.02±1.96, t = 2.6223, P 〈 0.05）. In the control group, the structure of liver lobule was normal and a few of collagen fibers loeated in portal area and central vein. In the 20 weeks hepatocirrhosis group, the structure of liver lobule was in disorder, a lot of hepatic eels were of fatty degeneration, and collagen fibers hyperplasia evidently. The immunohistochemistry staining of GRP78 in 20 weeks hepatoeirrhosis group was stronger than that of control group, and brown granules mostly loeate in the hepatic cytoplasm. Western blotting and RT-PCR also showed that the expression of GRP78 was enhanced. Conclusion In carbon tetrachloride induced hepatoeirrhosis, the expression of GRP78 increased evidently in liver. It demonstrated that hepatocirrhosis was in contact with endoplasmic reticulum stress.
出处
《解剖学报》
CAS
CSCD
北大核心
2006年第3期290-293,共4页
Acta Anatomica Sinica
基金
国家自然科学基金资助项目(30170244
30471638)
关键词
肝硬化
葡萄糖调节蛋白78
内质网应激
四氯化碳
免疫印迹法
大鼠
Hepatocirrhosis
Glucose regulating protein 78 （GRP78）
Endoplasmic reticulum stress
Carbon tetrachloride
Western blotting
Rat
作者简介
王晓红（1981-）,女（汉族）,山西省人,硕士研究生.
张卫光,通讯作者 E-mail：zhangook@yahoo.com.cn Tel：（010）82801466
