Journal Article

Chrysosplenetin acts as a homeostasis stabilizer with dual-function in shattering Plasmodium berghei K173 resistance to artemisinin driven by both ABC transporters and heme-ROS/GSH axis

Heng, Xin, Wang, Xiangyu, Yang, Junyi, Tan, Qingfeng, Chen, Linwei, Huang, Yifan, Bo, Ruonan, Tian, Jingxuan, Ji, Hongyan, Huang, Yutao, Gao, Hua, Chen, Jing

2025

Overview

Background Chrysosplenetin (CHR), a polymethoxy flavonol co-occurring with artemisinin (ART) in Artemisia annua L., reverses ART resistance in Plasmodium berghei K173 potentially by downregulating intestinal P-glycoprotein (P-gp, encoded by  Mdr1a ) expression. In the present study, we further elaborated on the mechanism by comparing differences in antimalarial activity and resistance-associated molecular expression profiles between ART alone and combination therapy in blood and tissues of Mdr1a wild-type (WT) and knockout (KO) mice infected with either sensitive or resistant malarial parasites. Methods We evaluated the effects of monotherapy and combination therapy in WT and KO mice infected with sensitive and resistant P. berghei K173 strains. The mRNA expressions of multi-resistance proteins (Mrp1, 2, 4, 5) and breast cancer resistance proteins (Bcrp) were detected. Hemoglobin levels, mRNA expressions of cytokines including tumor necrosis factor-α (IFN-α), interferon-α (IFN-α), and interleukin (IL-1β) in blood and tissues, and redox balance (ROS/GSH levels), as well as gene or protein expression of signaling pathway (PI3K/AKT-mTOR and MAPK) were investigated. Results In drug-resistant mice, combination therapy maintained the highest survival (100%) and inhibition (30%) rates and the lowest parasitaemia percentage (approximately 20.0%), irrespective of Mdr1a gene status. Furthermore, combination reshaped the spatial and ART resistance-phenotypic disparities in Mrps and Bcrp mRNA expressions (with a fold change ranging from 1.35 to 38.03), ROS/GSH balance (ranging from 1.02-fold to 10.18-fold), hemoglobin levels (ranging from 1.04-fold to 1.20-fold), and cytokine profiles (ranging from 1.14-fold to 37.79-fold) induced by ART alone, which were partially dysregulated by Mdr1a deficiency. Monotherapy and combination exert oppositely regulatory effects on the PI3K/AKT-mTOR pathway in a tissue-, Mdr1a genotype-, and parasite sensitivity/resistance-dependent manner (ranging from 1.52-fold to 84.00-fold). Specifically, CHR reversed ART-induced changes via PI3K/AKT protein inhibition (ranging from 1.20-fold to 63.00-fold), which was contingent on P-gp functionality. Finally, mitogen-activated protein kinase (MAPK) pathway was involved in the antagonistic regulation between ART alone and combination therapy in a P-gp-independent manner (ranging from 1.39-fold to 16.69-fold). Conclusions The efflux pump function of P-gp is probably not a critical factor in the mechanism by which CHR reverses ART resistance. Instead, CHR acts as a homeostasis stabilizer with dual functions: it disrupts Plasmodium berghei K173 resistance to ART driven by both ABC transporters and the heme-ROS/GSH axis, in which the non-transport function of P-gp on ART is involved. Graphical Abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: 1-Phosphatidylinositol 3-kinase / ABC transporters / AKT protein / Amino acids / Animals / Antibodies / Antimalarial activity / Antimalarials - pharmacology / Antiparasitic agents / Artemisia annua / Artemisinin / Artemisinin resistance / Artemisinins - pharmacology / Artificial intelligence / ATP-Binding Cassette Transporters - genetics / ATP-Binding Cassette Transporters - metabolism / Biological response modifiers / Biomedical and Life Sciences / Biomedicine / Blood / Blood parasites / breast neoplasms / Chrysosplenetin / Cloning / Cytokines / Drug resistance / Drug Resistance - drug effects / Drugs / Efflux / Entomology / Ethylenediaminetetraacetic acid / Female / Flavonols / Genes / Genotypes / Glutathione / Glutathione - metabolism / Glycoproteins / Haemoglobin / Heme / Heme - metabolism / Heme-ROS/GSH axis / Hemoglobin / Homeostasis / Homeostasis - drug effects / Infectious Diseases / interleukins / intestines / Kinases / Malaria / Malaria - drug therapy / Malaria - parasitology / MAP kinase / Medical research / Metabolites / Mice / Mice, Knockout / mitogen-activated protein kinase / mRNA / Multidrug resistance / Necrosis / NMR / Nuclear magnetic resonance / P-Glycoprotein / P-glycoproteins / Parasite resistance / parasitemia / Parasites / Parasitic diseases / Parasitology / PI3K/AKT-mTOR and MAPK pathways / Plasmodium berghei / Plasmodium berghei - drug effects / Plasmodium berghei - genetics / Protein folding / Protein kinases / protein synthesis / Proteins / Reactive oxygen species / Reactive Oxygen Species - metabolism / RNA / Signal transduction / Signal Transduction - drug effects / Sodium / stabilizers / therapeutics / Therapy / Tissue / TOR protein / Tropical Medicine / Veterinary Medicine/Veterinary Science / Virology / α-Interferon