獐牙菜苦苷通过促进 AMPK/mTOR 通路介导的自噬来缓解环磷酰胺引起的骨髓抑制。

Asperuloside alleviates cyclophosphamide-induced myelosuppression by promoting AMPK/mTOR pathway-mediated autophagy.

机构信息

Department of Hematology, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.

出版信息

J Biochem Mol Toxicol. 2024 Feb;38(2):e23641. doi: 10.1002/jbt.23641.

Abstract

Cyclophosphamide (CTX) is a common anticancer chemotherapy drug, and myelosuppression is the most common serious side effect. Asperuloside (ASP), the active component of Hedyotis diffusa Willd., may have the effect of ameliorating chemotherapy-induced myelosuppression. This study aimed to explore the effect and possible mechanism of ASP on CTX-induced myelosuppression. Male SPF C57BL/6 mice were randomly divided into five groups: control group, CTX (25 mg/kg) group, CTX + granulocyte-macrophage-colony stimulating factor (GM-CSF) (5 μg/kg) group, CTX + high-dose ASP (50 mg/kg) group and CTX + low-dose ASP (25 mg/kg) group, with six mice in each group. The body weight of mice was monitored every other day, the hematopoietic progenitor cell colony number was measured by colony forming unit, and the relevant blood indicators were detected. Femoral bone marrow was observed by hematoxylin-eosin, C-kit expression was detected by immunohistochemistry, and autophagy and adenine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway protein expressions were detected by immunohistochemistry and western blotting (WB). Then the AMPK inhibitor dorsomorphin was used to interfere with AMPK/mTOR pathway. Results showed that ASP significantly increased the body weight of CTX-induced mice, increased the number of hematopoietic progenitor cells, the expression of white blood cells, red blood cells, platelets, GM-CSF, thrombopoietin and erythropoietin in blood, and the expression of C-kit in bone marrow. In addition, ASP further promoted the expression of Beclin1 and LC-3II/I induced by CTX, and regulated the protein expressions in the AMPK/mTOR pathway. The use of dorsomorphin inhibited the alleviation effect of ASP on CTX-induced myelosuppression and the promotion effect of ASP on autophagy. In conclusion, ASP alleviated CTX-induced myelosuppression by promoting AMPK/mTOR pathway-mediated autophagy.

摘要

环磷酰胺（CTX）是一种常见的抗癌化疗药物，骨髓抑制是最常见的严重副作用。白花蛇舌草中的活性成分熊果酸（ASP）可能具有改善化疗引起的骨髓抑制的作用。本研究旨在探讨 ASP 对 CTX 诱导的骨髓抑制的作用及可能的机制。雄性 SPF C57BL/6 小鼠随机分为 5 组：对照组、CTX（25mg/kg）组、CTX+粒细胞-巨噬细胞集落刺激因子（GM-CSF）（5μg/kg）组、CTX+高剂量 ASP（50mg/kg）组和 CTX+低剂量 ASP（25mg/kg）组，每组 6 只。每隔一天监测小鼠体重，集落形成单位法检测造血祖细胞集落数，检测相关血液指标。苏木精-伊红染色观察股骨骨髓，免疫组化法检测 C-kit 表达，免疫组化和 Western blot（WB）法检测自噬和腺苷单磷酸激活蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）通路蛋白表达。然后用 AMPK 抑制剂 dorsomorphin 干扰 AMPK/mTOR 通路。结果表明，ASP 显著增加了 CTX 诱导的小鼠体重，增加了造血祖细胞的数量，增加了血液中白细胞、红细胞、血小板、GM-CSF、血小板生成素和促红细胞生成素的表达，以及骨髓中 C-kit 的表达。此外，ASP 进一步促进了 CTX 诱导的 Beclin1 和 LC-3II/I 的表达，并调节了 AMPK/mTOR 通路中的蛋白表达。使用 dorsomorphin 抑制了 ASP 对 CTX 诱导的骨髓抑制的缓解作用和 ASP 对自噬的促进作用。综上所述，ASP 通过促进 AMPK/mTOR 通路介导的自噬来缓解 CTX 诱导的骨髓抑制。