摘要
目的 观察RNA干扰肝脏Kupffer细胞肿瘤坏死因子-α（TNF-α）对大鼠肝脏缺血再灌注损伤的保护作用.方法 构建针对大鼠TNF-α基因的短发夹状RNA（shRNA）真核表达载体.肝脏缺血再灌注损伤前48 h经门静脉注射磷酸盐缓冲液（PBS）、空载体或TNF-α shRNA.实验随机分为4组,假手术组、PBS组、空载体组和shRNA组.阻断大鼠70%入肝血流40 min,再灌注6 h检测血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、肝脏Kupffer细胞TNF-α mRNA、血清TNF-α、肝组织中丙二醛（MDA）以及超氧化物岐化酶（SOD）含量.结果 与PBS组和空载体组比较,shRNA组再灌注6 h后血清ALT和AST水平显著降低（P＜0.05）,Kupffer细胞TNF-α mRNA水平、血清TNF-α水平（56.6±6.7 pg/ml比87.8±8.7 pg/ml和96.5±7.3 pg/ml,P＜0.05）、肝组织中MDA含量（93.4±13.3 nmol/mg比133.5±12.4 nmo1/mg和136.7±13.6 nmol/mg,P＜0.05）显著降低,SOD活性显著升高（22.4±4.6 U/mg比12.2±3.1 U/mg和11.4±2.9 U/mg,P＜0.05）.结论 RNA干扰Kupffer细胞TNF-α基因的表达可以减轻大鼠肝脏缺血再灌注损伤.
Objective To investigate the protective effect of silencing tumor necrosis factor-α （TNF-α） gene in Kupffer cells on liver ischemic reperfusion injury in rats. Methods We constructed the eukaryotic expression vector of small hairpin RNA targeting rat TNF-α gene. PBS, blank vector or TNF-α shRNA mixture were injected through portal vein 48 h before operation. The animals were divided into 4 groups： sham group, PBS group, blank vector group, and shRNA group. The left lateral and median lobes of the liver were rendered ischemic for 40 min resulting in a segmental （70 %） hepatic ischemia. The serum ALT and AST levels were determined 6 h after reperfusion. The mRNA expression of TNF-α in Kupffer cells, serum TNF-α level, concentration of the malondialdehyde （MDA） and superoxide dismutase（SOD） in the liver tissue were detected. Results Compared with the PBS and blank vector groups, ALT and AST levels were significantly decreased in the shRNA group after reperfusion for 6 h （P〈0. 05）, TNF-α mRNA level, serum TNF-α level（56.6 ± 6. 7 pg/ml vs 87.8 ± 8. 7 pg/ml and 96.5±7.3 pg/ml,P〈0. 05）, concentration of MDA in the liver tissue（93.4±13.3 nmol/mg vs 133.5±12. 4 nmol/mg and 136.7 ±13. 6 nmol/mg, P〈0.05 ）were significantly decreased, and SOD activity was significantly increased（22.4±4.6 U/mg vs 12. 2±3. 1 U/mg and 11. 4±2. 9 U/mg,P〈0. 05）. Conclusion RNA interfering TNF-α gene in Kupffer cells can protect the liver against ischemia/reperfusion injury.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2010年第10期767-769,共3页
Chinese Journal of Hepatobiliary Surgery
基金
教育部留学回国人员科研启动基金（2008-890号）
