Wu Zongmei, Geng Yana, Buist-Homan Manon, Moshage Han
Department of Gastroenterology and Hepatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
Department of Gastroenterology and Hepatology, University Medical Center Groningen, University of Groningen, Groningen, the Netherlands.
Toxicol Appl Pharmacol. 2022 Feb 1;436:115858. doi: 10.1016/j.taap.2021.115858. Epub 2021 Dec 31.
The number of patients with non-alcoholic fatty liver disease (NAFLD) is rapidly increasing due to the growing epidemic of obesity. Non-alcoholic steatohepatitis (NASH), the inflammatory stage of NAFLD, is characterized by lipid accumulation in hepatocytes, chronic inflammation and hepatocyte cell death. Scopoletin and umbelliferone are coumarin-like molecules and have antioxidant, anti-cancer and anti-inflammatory effects. Cytoprotective effects of these compounds have not been described in hepatocytes and the mechanisms of the beneficial effects of scopoletin and umbelliferone are unknown.
To investigate whether scopoletin and/or umbelliferone protect hepatocytes against palmitate-induced cell death. For comparison, we also tested the cytoprotective effect of scopoletin and umbelliferone against bile acid-induced cell death.
Primary rat hepatocytes were exposed to palmitate (1 mmol/L) or the hydrophobic bile acid glycochenodeoxycholic acid (GCDCA; 50 μmol/L). Apoptosis was assessed by caspase-3 activity assay, necrosis by Sytox green assay, mRNA levels by qPCR, protein levels by Western blot and production of reactive oxygen species (ROS) by fluorescence assay.
Both scopoletin and umbelliferone protected against palmitate and GCDCA-induced cell death. Both palmitate and GCDCA induced the expression of ER stress markers. Scopoletin and umbelliferone decreased palmitate- and GCDCA-induced expression of ER stress markers, phosphorylation of the cell death signaling intermediate JNK as well as ROS production.
Scopoletin and umbelliferone protect against palmitate and bile acid-induced cell death of hepatocytes by inhibition of ER stress and ROS generation and decreasing phosphorylation of JNK. Scopoletin and umbelliferone may hold promise as a therapeutic modality for the treatment of NAFLD.
由于肥胖症的流行日益严重，非酒精性脂肪性肝病（NAFLD）患者的数量正在迅速增加。非酒精性脂肪性肝炎（NASH）是NAFLD的炎症阶段，其特征是肝细胞内脂质堆积、慢性炎症和肝细胞死亡。东莨菪素和伞形酮是香豆素类分子，具有抗氧化、抗癌和抗炎作用。这些化合物对肝细胞的细胞保护作用尚未见报道，东莨菪素和伞形酮有益作用的机制也尚不清楚。
研究东莨菪素和/或伞形酮是否能保护肝细胞免受棕榈酸诱导的细胞死亡。为作比较，我们还测试了东莨菪素和伞形酮对胆汁酸诱导的细胞死亡的细胞保护作用。
原代大鼠肝细胞暴露于棕榈酸（1 mmol/L）或疏水性胆汁酸甘氨鹅去氧胆酸（GCDCA；50 μmol/L）。通过半胱天冬酶-3活性测定评估细胞凋亡，通过Sytox green测定评估细胞坏死，通过qPCR测定mRNA水平，通过蛋白质印迹测定蛋白质水平，通过荧光测定评估活性氧（ROS）的产生。
东莨菪素和伞形酮均能保护肝细胞免受棕榈酸和GCDCA诱导的细胞死亡。棕榈酸和GCDCA均诱导内质网应激标志物的表达。东莨菪素和伞形酮可降低棕榈酸和GCDCA诱导的内质网应激标志物的表达、细胞死亡信号中间体JNK的磷酸化以及ROS的产生。
东莨菪素和伞形酮通过抑制内质网应激和ROS生成以及减少JNK磷酸化来保护肝细胞免受棕榈酸和胆汁酸诱导的细胞死亡。东莨菪素和伞形酮有望成为治疗NAFLD的一种治疗方式。
