急性肝损伤小鼠中线粒体解耦联蛋白2的表达变化及意义

目的:观察C57小鼠急性肝损伤(acute liver injury,ALI)中解偶联蛋白2(Uncouple Protein2,UCP2)的表达,探讨ALI与UCP2表达变化的意义。方法:领取36只小鼠,随机分为对照组、ALI 1 d组、ALI 4 d组、ALI 7 d组。分别检测血清谷氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)的变化;肝组织病理变化用HE染色观察;利用Western Blot方法分别检测全肝组织蛋白和肝细胞线粒体提纯蛋白中的UCP2蛋白水平的变化;Real Time quality PCR检测UCP2在mRNA水平的表达变化。结果:ALI组1 d、4 d组与对照组相比,ALT(160.69±22.11 vs 34.43±5.19;96.37±15.39 vs 34.43±5.19)、AST(306.54±68.09 vs 97.74±14.49;173.94±26.74 vs 97.74±14.49)表达差异具有统计学意义(P0.05);肝组织病理学检查ALI组1 d和4 d组中肝细胞出现大面积坏死,7d组肝细胞坏死缓解;蛋白水平检测UCP2在ALI 1 d和4 d时全肝组织分别增加2.84倍和2.25倍,在肝线粒体中1 d、4 d和7 d时分别增加2.19倍、1.68倍和1.56倍,差异具有统计学意义(P0.05);mRNA水平检测UCP2在ALI 1 d和4 d与对照组相比明显升高,相对增加3.79倍和1.46倍(P0.05)。结论:急性肝损伤状态下UCP2在蛋白和mRNA水平高表达,UCP2可能对于治疗急性肝损伤具有重要意义。

Expression of Uncoupling Protein 2 in Mice with Acute Liver Injury and its Significance

ABSTRACT Objective: To observe the change of uncoupling protein 2 (UCP2) and its correlation with liver injury in the C57/BL mice with thioacetamide (TAA) induced acute liver injury (ALI). Methods: C57/BL mice were randomly divided into four groups: control group, ALI 1 d, ALI 4 d and ALI 7 d. Mice received intra-abdominal injection of TAA (300 mg?kg-1?d-1 for 3 days) in the ALI groups, and the control group was injected equal volume of 0.9 % NaCl. The level of serum aspartate amino transferase (AST) and alanine aminotransferase (ALT) in serum was detected by Elisa. The samples of liver tissue were harvested and pathomorphological changes of samples were examined by hematoxylin-eosin (HE) staining. Western Blot was used to detect the expression of UCP2 protein. Real time quality PCR was used to evaluate the expression of UCP2 in the mRNA level. Results: The level of ALT (160.69±22.11 vs 34.43±5.19; 96.37±15.39 vs 34.43±5.19)and AST (306.54±68.09 vs 97.74±14.49; 173.94±26.74 vs 97.74±14.49) in ALI groups were signifi- cantly higher than those in the control group at 1 d and 4 d (P<0.05), and peaked at 1d; HE staining showed that a mass of inflammatory cells infiltration and cells degeneration were observed in ALI groups, and the damage was evident at 1 d and 4 d; Western Blot detected that the expression of UCP2 in the extracted protein of liver tissue and liver mitochondria increased significantly at 1 d (2.84 and 2.19 - fold, respectively) and 4 d (2.25 and 1.68 - fold, respectively) compared with control group (P<0.05); The result of real time quality-PCR indicated that the level of UCP2 enhanced significantly at 1 d (3.79 - fold) and 4 d (1.46 - fold) in ALI groups compared with control group (P<0.05). Conclusion: UCP2 was highly expressed in protein and mRNA levels in acute liver injury, and UCP2 could be important for the treatment of acute liver injury.