NATURAL PRODUCT RESEARCH AND DEVELOPMENT ›› 2023, Vol. 35 ›› Issue (4): 573-583.doi: 10.16333/j.1001-6880.2023.4.003

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Toddalia asiatica promotes lipopolysaccharide-induced autophagy and inhibits inflammation in RAW 264.7 cells based on the PI3K/AKT/mTOR pathway

ZHANG Zong-xing1,2,JIANG Lu1,2,LIU Dao-zhong1,2,HOU Zi-ming1,2,TIAN Meng-jie1,2,TAO Bo-nan1,2,FENG Jia1,2,YUAN Lin1,2*   

  1. 1Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University;2Department of Medicine,Hubei Minzu University,Enshi 445000,China
  • Online:2023-04-24 Published:2023-04-24

Abstract:

The root of the Rutaxaceae plant Toddalia asiatica (L.) Lam is a natural Tujia Chinese herbal medicine with anti-inflammatory,anti-rheumatic,anti-tumor,anti-microbial and other pharmacological activities.Its characteristics of small poisonous side effects and remarkable curative effect make it a hot spot in current research.Many natural products had been shown to inhibit inflammation and autoimmune diseases by targeting PI3K/AKT/mTOR mediated autophagy.In this study,the effect of TAAE on autophagy was studied by regulating PI3K/AKT/mTOR signaling pathway,and RAW 264.7 cells were induced by LPS to establish an inflammatory model.Cytotoxicity detection kit was used to detect the effect of TAAE on cell viability,the concentration and intervention time of the drug were screened out.Transmission electron microscopy and MDC staining were used to detect the biological function of macrophages.The levels of inflammatory factors in the supernatant were detected by ELISA,and the expression levels of autophagy and pathway-related proteins were detected by Western blot.The effects of autophagy on inflammation and signaling pathways were further verified by using an early autophagy inhibitor (3-MA) and pathway PI3K agonist (740Y-P).The results indicated that TAAE may inhibit the PI3K/AKT/mTOR signaling pathway,promote the formation of autophagic vesicles,fusion and degradation of autophagosomal lysosomes,and reduce the expression and secretion of inflammatory cytokines in RAW 264.7 cells treated with LPS.Overall,the results of this study provide a new clue for the research of the anti-inflammatory mechanism of Toddalia asiatica,and provide a theoretical basis for better clinical application of Toddalia asiatica in the treatment of inflammatory diseases.

Key words: Toddalia asiatica, PI3K/AKT/mTOR pathway, autophagy, inflammation, RAW 264.7 cells

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