Zhang Nan, Sheng Mingwei, Wu Man, Zhang Xinyue, Ding Yijie, Lin Yuanbang, Yu Wenli, Wang Shusen, Du Hongyin
Department of Anesthesiology, Tianjin First Central Hospital, Tianjin 300192, China.
Department of Anesthesiology, China-Japan Friendship Hospital, Beijing 100029, China.
Exp Biol Med (Maywood). 2019 Dec;244(18):1695-1704. doi: 10.1177/1535370219878651. Epub 2019 Sep 25.
Steatotic livers are more susceptible to ischemia/reperfusion injury, and increase the risk of primary graft non-function after liver transplantation. The protective effects of berberine have been described in various liver pathological models. However, it is unknown if berberine exerts its beneficial action in steatotic donors undergoing liver transplantation. In the present study, male Wistar rats were fed with high-fat diet (HFD) for 12 weeks to induce moderate steatotic liver. Then orthotropic liver transplantation was constructed. Berberine (200 mg/kg/d) was given intragastrically one week before liver transplantation. Thapsigargin (TG) (0.2 mg/kg) was administrated intravenously 24 h before liver transplantation. Liver function, oxidative stress, and inflammatory cytokine were detected by biochemical or histopathological analysis. The morphology of autophagosomes and endoplasmic reticulum (ER) was observed by transmission electron microscopy. The expression of CHOP, BIP, the phosphorylation of PERK, LC3-II/I, Beclin-1, and p62 were determined by Western blot assay. The co-localization of endoplasmic reticulum marker (KDEL) and autophagic protein (LC3B) was analyzed by immunofluorescence microscopy. The level of reticulophagy hallmark (FAM134B) was determined by immunohistochemistry. Compared with HFD + LT group, berberine ameliorated hepatocellular damage, decreased the oxidative stress level and inflammatory cytokine release. Simultaneously, berberine inhibited the expression of both endoplasmic reticulum stress parameters and autophagy-related proteins. Additionally, the co-localization of endoplasmic reticulum marker and LC3B was also reduced in HFD + BBR + LT group. berberine down-regulated the level of FAM134B. TG reversed the beneficial effects of berberine. Our study revealed that berberine exerts protective effects on steatotic livers undergoing transplantation by inhibiting endoplasmic reticulum stress-mediated reticulophagy.
Berberine is isolated from traditional Chinese medicine plants and has dramatically therapeutic potential against inflammation, diarrhea, and diabetes. But the benefits of BBR on steatotic grafts after liver transplantation remain poorly understood. Our findings might help explain the mechanism of berberine in protecting steatotic livers undergoing transplantation and give advantageous insights that berberine has potential as a suitable candidate for preventing hepatic injury after steatotic liver transplantation by inhibiting ER stress-mediated reticulophagy.
脂肪变性的肝脏对缺血/再灌注损伤更敏感，并增加肝移植后原发性移植物无功能的风险。黄连素在各种肝脏病理模型中均已显示出保护作用。然而，黄连素在接受肝移植的脂肪变性供体中是否发挥有益作用尚不清楚。在本研究中，雄性Wistar大鼠喂食高脂饮食（HFD）12周以诱导中度脂肪变性肝脏。然后构建原位肝移植模型。在肝移植前一周，通过灌胃给予黄连素（200mg/kg/d）。在肝移植前24小时静脉注射毒胡萝卜素（TG）（0.2mg/kg）。通过生化或组织病理学分析检测肝功能、氧化应激和炎性细胞因子。通过透射电子显微镜观察自噬体和内质网（ER）的形态。通过蛋白质免疫印迹法检测CHOP、BIP、PERK磷酸化、LC3-II/I、Beclin-1和p62的表达。通过免疫荧光显微镜分析内质网标志物（KDEL）和自噬蛋白（LC3B）的共定位。通过免疫组织化学测定网织自噬标志（FAM134B）的水平。与HFD+LT组相比，黄连素改善了肝细胞损伤，降低了氧化应激水平和炎性细胞因子释放。同时，黄连素抑制内质网应激参数和自噬相关蛋白的表达。此外，HFD+BBR+LT组内质网标志物与LC3B的共定位也降低。黄连素下调了FAM134B的水平。TG逆转了黄连素的有益作用。我们的研究表明，黄连素通过抑制内质网应激介导的网织自噬对移植的脂肪变性肝脏发挥保护作用。
黄连素是从传统中药植物中分离出来的，对炎症、腹泻和糖尿病具有显著的治疗潜力。但黄连素对肝移植后脂肪变性移植物的益处仍知之甚少。我们的研究结果可能有助于解释黄连素保护移植脂肪变性肝脏的机制，并提供有益的见解，即黄连素有可能通过抑制内质网应激介导的网织自噬，成为预防脂肪变性肝移植后肝损伤的合适候选药物。
