Abstract:

Objective To investigate the protective effect of berberine pretreatment on liver injury in rats undergoing autogenous orthotopic liver transplantation and its related mechanism. Methods Twenty-four cleangrade healthy male Sprague-Dawley rats were divided into 3 groups（n ＝ 8）. They were sham operation group（S group），model group（AT group）and berberine pretreatment group（B group）. The weight was 250 ～ 280 g. In S group，the abdomen was opened，the corresponding blood vessels and ligaments were isolated，and the abdomen wasclosed. Rat models of liver injury after orthotopic liver transplantation were prepared in both AT and B groups，andberberine〔200 mg/（kg·d）〕was given by gavage 1 week before surgery in B group. Rats were anesthetized at 6 h after reperfusion，and the serum and liver tissues were collected. The serum levels of alanine aminotransferase（ALT）and aspartate aminotransferase（AST）were detected，the contents of serum high mobility group box-1 protein（HMGB1）（by enzyme-linked immunosorbent assay），and the expression of peroxisome proliferator activated receptor（PPARγ） and nuclear transcription factor-κB（NF-κB）（by immunohistochemistry）were determined. The expressions of ASC，NLRP3 and Pro-caspase1（by Western blot）were detected. Results Compared with S group，the levelsof serum ALT and AST and the contents of HMGB1 were significantly increased. The expression of PPARγin liver was down-regulated and the expression NF-κB、ASC、NLRP3 and Pro-caspase1 in liver was up-regulated，and histopathological injury of liver was significantly aggravated in AT and B groups（P ＜ 0.05）. Compared with AT group，the levels of serum ALT and AST and the contents of HMGB1 were significantly decreased. The expression of PPARγin liver was up-regulated and the expression NF-κB、ASC、NLRP3 and Pro-caspase1 in liver wasdown-regulated in liver tissue（P ＜ 0.05），and liver histopathological damage was significantly improved in B group. Conclusion Pretreatment of berberine can inhibit the expression of NF-κB by activating PPARγand then inhibiting the pyroptosis pathway，thus play a protective role against liver injury induced by liver transplantation in rats. 

Key words:

Liver transplantation , Liver injury , Berberine , Peroxisome proliferator activated receptorγ, Nuclear transcription factor-κB, Pyroptosis

摘要：

目的 探讨小檗碱预处理在大鼠自体原位肝移植术诱发肝脏损伤的保护作用及相关的机制。 方法 健康雄性清洁级 SD 大鼠 24 只，体重为 250 ～ 280 g，分为 3 组（n ＝ 8）：假手术组（S 组）、模型组（AT 组）和小檗碱预处理组（B 组）。S 组仅进行开腹，游离肝脏周围的血管和韧带，关腹。AT 组和 B 组均制备自体原位肝移植诱发肝脏损伤大鼠模型，B 组术前 1 周灌胃小檗碱〔200 mg/（kg·d）〕。在灌注 6 h 时麻醉大鼠，取其血清和肝脏组织，检测血清丙氨酸转氨酶（alanine transaminase，ALT）和天冬氨酸转氨酶（aspartate transaminase，AST）水平，采用酶联免疫吸附试验（enzyme-linked immunosorbent assay，ELISA）法测定血清高迁移率族蛋白 B-1（high mobility group box-1 protein，HMGB-1）含量，采用免疫组织化学法测定肝脏组织过氧化物酶体增殖物激活受体 γ（peroxisome proliferators-activated receptors γ，PPARγ）和核转录因子 -κB（nuclear transcription factor-κB，NF-κB）表达，采用蛋白免疫印迹法检测凋亡相关斑点样蛋白（apoptosis associated spot-like protein，ASC）、NLRP3 和Pro-caspase1 蛋白表达。 结果 与 S 组比较，AT 组血清 ALT 、AST 水平和 HMGB1 含量升高。肝脏组织 PPARγ 表达下调，NF-κB、ASC、NLRP3 和 Pro-caspase1 表达上调（P ＜ 0.05），肝脏组织病理学损伤明显加重。与 AT 组比 较，B 组血清 ALT 、AST 水平和 HMGB1 含量降低，肝脏组织 PPARγ 表达上调，NF-κB、ASC、NLRP3 和 Pro-caspase1 表达下调（P ＜ 0.05），肝脏组织病理学损伤明显改善。 结论 预处理小檗碱可以通过激活 PPARγ，抑制 NF-κB 表达，进而抑制细胞焦亡通路，从而发挥对大鼠肝脏移植过程诱发肝损伤的保护作用。

关键词:

肝移植 , 肝脏损伤 , 小檗碱 , 过氧化物酶体增殖物激活受体 γ , 核转录因子 -κB , 细胞焦亡