Tian Mengxi, Liu Wenlan, You Hongjie, Zhao Qingzhou, Ouyang Luodan, Gao Biane, Zhang Xin, Che Niancong
J Tradit Chin Med. 2016 Aug;36(4):471-8. doi: 10.1016/s0254-6272(16)30064-4.
To investigate the inhibitory effect of Yiguanjian decoction (YD) on DNA damage in Concanavalin A (Con A)-induced liver injury mice model and to explain the possible mechanism.
METHODS: Totally 120 male BALB/c mice were randomly divided into 6 groups, 20 mice each: normal group, model group, Bifendate group, YD low dose group, YD middle dose group and YD high dose group. Except normal group, liver injury model induced by Con A was established. While modeling, each mouse in YD group was given YD (0.4 mL/20 g per day) by intragastric administration (0.13 g YD for YD low dose group; 0.26 g for YD middle dose group; 0.52 g for YD high dose group). Bifendate group was given Bifendate (0.2 g·kg-1·d-1) by gavage. Normal group and model group were fed with same volume of physiological saline daily. After 8 weeks, the serum alanine transaminase (ALT) and aspartate transaminase (AST) were tested. The hematoxylin-eosin staining was used to evaluate the grade of liver inflammation and liver fibrosis stage. Hepatocellular DNA damage was detected by single cell gel electrophoresis technology. The protein expression of tumor necrosis factor-α (TNF-α), Bax and MutT Homolog 1 (MTH1) was detected by western blotting and enzyme linked immunosorbent assay. Bax mRNA and MTH1 mRNA were detected by Real-time Polymerase Chain Reaction (PCR).
YD can improve the degree of liver inflammation and fibrosis in the liver of chronic hepatitis mice, the dose effect relationship is remarkable (P < 0.05). YD can reduce liver cell DNA damage. The difference between YD middle dose group and model group was statistically significant (P < 0.05). YD middle dose group had decreased the protein expression of TNF-α in the mice liver of immunological liver injury (P < 0.05). YD can increase the protein expression of Bax (P < 0.05). Compared with normal group, the protein expression of MTH1 was decreased (P < 0.05), but there was no statistical significance between YD group and model group (P > 0.05). YD can increase the mRNA expression of Bax and MTH1 (both P < 0.05).
YD can effectively inhibit the DNA damage in immunological liver injury mice, the mechanism may be that it can decrease the TNF-α and increase the Bax and MTH1 expression.
探讨一贯煎（YD）对刀豆蛋白A（Con A）诱导的肝损伤小鼠模型DNA损伤的抑制作用，并阐明其可能机制。
将120只雄性BALB/c小鼠随机分为6组，每组20只：正常组、模型组、联苯双酯组、YD低剂量组、YD中剂量组和YD高剂量组。除正常组外，其余各组建立Con A诱导的肝损伤模型。造模期间，YD组小鼠每日按0.4 mL/20 g灌胃给予YD（YD低剂量组为0.13 g YD；中剂量组为0.26 g；高剂量组为0.52 g）。联苯双酯组按0.2 g·kg-1·d-1灌胃给予联苯双酯。正常组和模型组每日给予等体积生理盐水。8周后，检测血清丙氨酸转氨酶（ALT）和天冬氨酸转氨酶（AST）。采用苏木精-伊红染色评估肝脏炎症程度和肝纤维化分期。采用单细胞凝胶电泳技术检测肝细胞DNA损伤。采用蛋白质印迹法和酶联免疫吸附测定法检测肿瘤坏死因子-α（TNF-α）、Bax和MutT同源蛋白1（MTH1）的蛋白表达。采用实时聚合酶链反应（PCR）检测Bax mRNA和MTH1 mRNA。
YD可改善慢性肝炎小鼠肝脏炎症和纤维化程度，剂量效应关系显著（P<0.05）。YD可减轻肝细胞DNA损伤。YD中剂量组与模型组比较差异有统计学意义（P<0.05）。YD中剂量组可降低免疫性肝损伤小鼠肝脏中TNF-α的蛋白表达（P<0.05）。YD可增加Bax的蛋白表达（P<0.05）。与正常组比较，MTH1的蛋白表达降低（P<0.05），但YD组与模型组比较差异无统计学意义（P>0.05）。YD可增加Bax和MTH1的mRNA表达（均P<0.05）。
YD可有效抑制免疫性肝损伤小鼠的DNA损伤，其机制可能是降低TNF-α水平，增加Bax和MTH1表达。
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