School of Pharmaceutical Sciences, South-Central Minzu University, Wuhan, China.
School of Life Sciences, Nanjing University, Nanjing, China.
J Ethnopharmacol. 2024 Mar 1;321:117495. doi: 10.1016/j.jep.2023.117495. Epub 2023 Nov 26.
When left untreated, liver fibrosis (LF) causes various chronic liver diseases. Earthworms (Pheretima aspergillum) are widely used in traditional medicine because of their capacity to relieve hepatic diseases.
This study aimed to explore the anti-LF effects of water extract of earthworms (WEE) and the underlying molecular mechanisms.
A CCl-induced mouse model of LF was used to study the impact of WEE on LF in vivo. The anti-LF activity of WEE in mice was compared with that of silybin, which can be clinically applied in LF intervention and was used as a positive control. Activation of LX-2 hepatic stellate cells (HSCs) and apoptosis and ferroptosis of AML-12 hepatocytes induced by TGFβ1 were used as in vitro models.
WEE drastically improved LF in mice. WEE reduced markers of activated HSCs in mice and inhibited TGFβ1-induced activation of LX-2 HSCs in vitro. Additionally, WEE suppressed CCl-induced apoptosis and ferroptosis in mouse hepatocytes. Mechanistically, WEE induced Nrf2 to enter the nuclei of the mouse liver cells, and the hepatic levels of Nrf2-downstream antioxidative factors increased. LKB1/AMPK/GSK3β is an upstream regulatory cascade of Nrf2. In the LF mouse model, WEE increased hepatic phosphorylated LKB1, AMPK, and GSK3β levels. Similar results were obtained for the LX-2 cells. In AML-12 hepatocytes and LX-2 HSCs, WEE elevated intracellular Nrf2 levels, promoted its nuclear translocation, and inhibited TGFβ1-induced ROS accumulation. Knocking down LKB1 abolished the impact of WEE on the AMPK/GSK3β/Nrf2 cascade and eliminated its protective effects against TGFβ1.
Our findings reveal that WEE improves mouse LF triggered by CCl and supports its application as a promising hepatoprotective agent against LF. The potentiation of the hepatic antioxidative AMPK/GSK3β/Nrf2 cascade by activating LKB1 and the subsequent suppression of HSC activation and hepatocyte apoptosis and ferroptosis are implicated in WEE-mediated alleviation of LF.
未经治疗，肝纤维化（LF）会导致各种慢性肝病。蚯蚓（参环毛蚓）因其缓解肝脏疾病的能力而在传统医学中广泛应用。
本研究旨在探讨蚯蚓水提物（WEE）的抗 LF 作用及其潜在的分子机制。
采用 CCl 诱导的 LF 小鼠模型研究 WEE 对 LF 的体内影响。将 WEE 的抗 LF 活性与临床上可用于 LF 干预的水飞蓟宾进行比较，水飞蓟宾作为阳性对照。激活 TGFβ1 诱导的 LX-2 肝星状细胞（HSCs）和 AML-12 肝细胞凋亡和铁死亡被用作体外模型。
WEE 显著改善了 LF 小鼠的病情。WEE 降低了小鼠活化 HSCs 的标志物，并抑制了 TGFβ1 诱导的 LX-2 HSCs 的体外活化。此外，WEE 抑制了 CCl 诱导的小鼠肝细胞凋亡和铁死亡。在机制上，WEE 诱导 Nrf2 进入小鼠肝细胞的核内，肝脏中 Nrf2 下游抗氧化因子的水平增加。LKB1/AMPK/GSK3β 是 Nrf2 的上游调节级联。在 LF 小鼠模型中，WEE 增加了肝组织中磷酸化 LKB1、AMPK 和 GSK3β 的水平。在 LX-2 细胞中也得到了类似的结果。在 AML-12 肝细胞和 LX-2 HSCs 中，WEE 提高了细胞内 Nrf2 水平，促进其核转位，并抑制 TGFβ1 诱导的 ROS 积累。敲低 LKB1 消除了 WEE 对 AMPK/GSK3β/Nrf2 级联的影响，并消除了其对 TGFβ1 的保护作用。
我们的研究结果表明，WEE 可改善 CCl 诱导的 LF 小鼠，并支持其作为一种有前途的抗 LF 肝保护剂的应用。通过激活 LKB1 增强肝脏抗氧化 AMPK/GSK3β/Nrf2 级联，随后抑制 HSC 活化和肝细胞凋亡和铁死亡，WEE 介导的 LF 缓解与这一机制有关。
