中文摘要：

目的建立经门静脉高通量注射将裸DNA质粒导入大鼠肝脏表达的实验方法,观察FasL在肝脏的表达、对肝损伤及肝功能生化指标变化的影响。方法对大鼠经门静脉高通量注射质粒pDC316-LacZ,观察LacZ基因在大鼠肝脏的表达情况。将11只大鼠随机分为实验组6只（注射pDC315-rFasL）,对照组5只（注射pDC316-LacZ）,经门静脉高通量注射将大鼠FasL真核质粒导入大鼠肝脏。分别于术后第1、3-4、6天处死大鼠,术前、术后取血查ALT和AST,肝组织进行RT-PCR、Western印迹及常规HE染色和免疫组织化学实验。结果经门静脉高通量注射了pDC316-LacZ的大鼠肝脏,可观察到蓝色沉淀物质在肝组织表达。经门静脉将FasL质粒导入大鼠肝脏,24h后取肝组织通过RT-PCR和Western印迹实验证实FasL在肝脏中表达。免疫组织化学结果表明在术后实验组肝组织大量表达FasL。常规HE染色显示实验组术后肝小叶中肝细胞嗜酸性变及凋亡小体明显高于对照组。术后第1天实验组与对照组ALT、AST水平显著升高。第3～4天对照组恢复正常,但实验组于术后第6天才基本正常。结论初步建立了门静脉高通量注射将裸DNA质粒导入大鼠肝脏表达的实验方法。肝细胞表达FasL可能成为效应细胞致邻近肝细胞凋亡。

英文摘要：

Objective To observe the overexpression of FasL in hepatic tissue, and the influence on liver injury, biochemical indicator alteration of liver function and expression of FasL, We develop the experimental method of high volume hydrodynamic injection of naked plasmid DNA via the rat portal vein. Methods Naked plasmid DNA of pDC316- LacZ was injected to rat liver in high volume hydrodynamic way via portal vein. We harvested the rat liver after 24 hours to detect the expression of LacZ gene with beta-galactosidase staining in situ. Eleven Wistar rats were divided into test group treated with pDC315-rFasL plasmid and control group treated with pDC316-LacZ with the method mentioned above. Rats were sacrificed on day 1, day 3/4, day 6 after surgery. Rat sera were harvested before and after surgery and the aminopherase was assayed. Rat livert issues were harvested for detecting expression of FasL by RT-PCR, Western blot, HE staining and immunohistochemistry. Results The expression of 13 galactosidase was found in rat liver injected with pDC316-LacZ plasmid. RT-PCR and Western blot on hepatic tissues harvested proved that FasL had been expressed in rat liver successfully. Immunohistochemistry results indicated that FasL had expressed abundantly in the hepatic lobule first region of rats liver in test group. The HE staining showed that the hepatocytes of test group appeared more acidophilia degeneration and apoptosis than those of control group. The serum transaminase increased significantly at day 1 in both groups. The level of transaminase recovered to normal on day 3/4 in control group, while that in test group recovered to normal on day 6. Conclusion High volume hydrodynamic injection of naked plasmid DNA via the hepatic portal vein has been initially developed. FasL-overexpressing hepatocytes may become effector cells to induce adjacent hepatocyteapoptosis.