Zhou J J, Qin J J, Lyu L, Zhou H M, Zhang F
Liver Transplantation Center of First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China.
Zhonghua Gan Zang Bing Za Zhi. 2018 Jan 20;26(1):42-47. doi: 10.3760/cma.j.issn.1007-3418.2018.01.010.
To investigate the role of short-term starvation (STS) in alleviating hepatic ischemia-reperfusion injury in mice and possible mechanism of action. Wild-type male C57BL/6 mice aged 8 weeks were randomly divided into 75% hepatic ischemia-reperfusion injury group (IR group), STS+75% hepatic ischemia-reperfusion injury group (STS group), and sirtinol+STS+75% hepatic ischemia-reperfusion injury group (SIR group), using a random number table, and sham-operation groups (IR-Sham group, STS-Sham group, and SIR-Sham group) were also established. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, and the histomorphological changes of the liver were observed, as well as the expression of Sirt1, LC3B, and P62 proteins in liver tissue and the results of LC3B fluorescence staining. An analysis of variance was used for comparison of data between multiple groups, and the t-test was used for comparison of data between two groups. Compared with the IR group, the STS group had significant reductions in the serum levels of ALT (3 152.7 ± 735.6 U/L vs 8 414.2 ± 1 052.2 U/L, < 0.01) and AST (3 577.0 ± 714.0 U/L vs 10 845.8 ± 1 145.7 U/L, < 0.01) and significant alleviation of liver pathological injury (Suzuki score: 1.50±0.55 vs 3.50±0.55, < 0.01). Compared with the STS group, the SIR group had significant increases in the serum levels of ALT (7 002.7 ± 1 485.2 U/L vs 3 152.7 ± 735.6 U/L, < 0.01) and AST (8 980.7 ± 1 739.1 U/L vs 3 577.0 ± 714.0 U/L, < 0.01) and significant exacerbation of liver pathological injury (Suzuki score: 3.33 ± 0.52 vs 1.50 ± 0.55, < 0.01). Compared with the IR group and the IR-Sham group, the STS group and the STS-Sham group had significant increases in the mRNA and protein expression of Sirt1 and the protein expression of LC3B and a significant reduction in the protein expression of P62, as well as a significant increase in the percentage of LC3B-positive cells in liver tissue (22.83% ± 5.19% / 22.17% ± 4.83% vs 10.16% ± 3.06% / 10.83% ± 1.94%, both < 0.01). Compared with the STS group and the STS-Sham group, the SIR group and the SIR-Sham group had significant reductions in the expression of Sirt1 and LC3B proteins and a significant increase in the expression of P62 protein, as well as a significant reduction in the percentage of LC3B-positive cells in liver tissue (11.83% ± 9.24% / 14.67% ± 4.68% vs 22.83% ± 5.19% / 22.17% ± 4.83%, both < 0.01). STS can effectively alleviate hepatic ischemia-reperfusion injury, and its protective effect may be associated with increasing the expression of Sirt1, inducing and promoting hepatocyte autophagy, and reducing hepatocyte death.
探讨短期饥饿（STS）在减轻小鼠肝脏缺血再灌注损伤中的作用及其可能的作用机制。将8周龄野生型雄性C57BL/6小鼠采用随机数字表法随机分为75%肝脏缺血再灌注损伤组（IR组）、STS + 75%肝脏缺血再灌注损伤组（STS组）、sirtinol + STS + 75%肝脏缺血再灌注损伤组（SIR组），并设立假手术组（IR-Sham组、STS-Sham组和SIR-Sham组）。检测血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平，观察肝脏组织形态学变化，以及肝脏组织中Sirt1、LC3B和P62蛋白的表达情况及LC3B荧光染色结果。多组间数据比较采用方差分析，两组间数据比较采用t检验。与IR组相比，STS组血清ALT水平（3152.7±735.6 U/L比8414.2±1052.2 U/L，P<0.01）和AST水平（3577.0±714.0 U/L比10845.8±1145.7 U/L，P<0.01）显著降低，肝脏病理损伤明显减轻（铃木评分：1.50±0.55比3.50±0.55，P<0.01）。与STS组相比，SIR组血清ALT水平（7002.7±1485.2 U/L比3152.7±735.6 U/L，P<0.01）和AST水平（8980.7±1739.1 U/L比3577.0±714.0 U/L，P<0.01）显著升高，肝脏病理损伤明显加重（铃木评分：3.33±0.52比1.50±0.55，P<0.01）。与IR组和IR-Sham组相比，STS组和STS-Sham组Sirt1的mRNA和蛋白表达、LC3B蛋白表达显著增加，P62蛋白表达显著降低，肝脏组织中LC3B阳性细胞百分比显著增加（22.83%±5.19%/22.17%±4.83%比10.16%±3.06%/10.83%±1.94%，均P<0.01）。与STS组和STS-Sham组相比，SIR组和SIR-Sham组Sirt1和LC3B蛋白表达显著降低，P62蛋白表达显著增加，肝脏组织中LC3B阳性细胞百分比显著降低（11.83%±9.24%/14.67%±4.68%比22.83%±5.19%/22.17%±4.83%，均P<0.01）。短期饥饿可有效减轻肝脏缺血再灌注损伤，其保护作用可能与增加Sirt1表达、诱导并促进肝细胞自噬、减少肝细胞死亡有关。
