State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing, China.
State Key Laboratory of Natural Medicines, Department of Biochemistry, China Pharmaceutical University, Nanjing, China.
Life Sci. 2021 Sep 15;281:119778. doi: 10.1016/j.lfs.2021.119778. Epub 2021 Jun 27.
Demethylenetetrahydroberberine (DMTHB) is a novel derivative of berberine and demethyleneberberine. This research explored the pharmacological effects and molecular mechanisms of DMTHB on nonalcoholic fatty liver disease (NAFLD).
C57BL/6 mice were induced by a methionine- and choline- deficient (MCD) diet and L02 cells were induced by palmitic acid to establish NAFLD animal and cell models. qPCR and western blotting were used to detect the expression of genes and proteins associated with pharmacological mechanism. A biotin-labeled DMTHB pulldown assay was used to further clarify the pharmacological targets.
Our results indicated that DMTHB significantly alleviates NAFLD in mice. Biochemical assays showed that serum alanine aminotransferase, aspartate aminotransferase and hepatic lipids were significantly decreased in MCD-induced NAFLD mice orally administered of DMTHB (50 mg/kg or 150 mg/kg body weight daily) for 30 d. qPCR and ELISA analysis demonstrated that DMTHB reduced the expression of serum proinflammatory cytokines, such as TNF-α, IL-1β and IL-6. Moreover, pull-down assays and compound-centric chemical proteomics illustrated that DMTHB inhibited NOD-like receptor protein 3 (NLRP3) inflammasome signaling. In addition, DMTHB also attenuated oxidative stress and endoplasmic reticulum stress by downregulation CYP2E-1 and ATF-4 expression. Moreover, DMTHB treatment ameliorated the liver fibrosis in MCD-induced NAFLD mice by suppressing the expression of TGF-β1, α-SMA and collagen 1A1.
DMTHB targeted the NLRP3 inflammasome to suppress inflammation and inhibited CYP2E1 to reduce oxidative stress and ER stress. Consequently, DMTHB may have therapeutic benefits in the treatment of NAFLD in the clinic.
脱氢小檗碱（DMTHB）是小檗碱和去甲小檗碱的一种新型衍生物。本研究探讨了 DMTHB 对非酒精性脂肪性肝病（NAFLD）的药理作用和分子机制。
采用蛋氨酸和胆碱缺乏（MCD）饮食诱导 C57BL/6 小鼠和棕榈酸诱导 L02 细胞建立 NAFLD 动物和细胞模型。采用 qPCR 和 Western blot 检测与药理机制相关的基因和蛋白表达。采用生物素标记的 DMTHB 下拉实验进一步阐明药理作用靶点。
我们的研究结果表明，DMTHB 可显著缓解 MCD 诱导的 NAFLD 小鼠的疾病。生化检测表明，经口给予 50mg/kg 或 150mg/kg 体重的 DMTHB （每日一次）30d 后，MCD 诱导的 NAFLD 小鼠血清丙氨酸转氨酶、天冬氨酸转氨酶和肝脂质显著降低。qPCR 和 ELISA 分析表明，DMTHB 降低了血清促炎细胞因子 TNF-α、IL-1β和 IL-6 的表达。此外，下拉实验和基于化合物的化学蛋白质组学表明，DMTHB 抑制了 NOD 样受体蛋白 3（NLRP3）炎症小体信号。此外，DMTHB 通过下调 CYP2E-1 和 ATF-4 的表达，减轻氧化应激和内质网应激。此外，DMTHB 通过抑制 TGF-β1、α-SMA 和胶原 1A1 的表达，改善 MCD 诱导的 NAFLD 小鼠的肝纤维化。
DMTHB 靶向 NLRP3 炎症小体抑制炎症，并抑制 CYP2E1 减少氧化应激和内质网应激。因此，DMTHB 可能在临床上对 NAFLD 的治疗具有治疗益处。
