Jianpi Xiao'ai Prescription Inhibits Colorectal Cancer Progression by Inducing Mitochondrial Dysfunction via Modulation of iNOS-ARG1 Axis
10.13422/j.cnki.syfjx.20251729
- VernacularTitle:健脾消癌方调控iNOS-ARG1轴诱导线粒体功能障碍抑制结肠癌进展
- Author:
Xing LUO
1
;
Bo PAN
2
;
Jianfeng FU
2
;
Jia HUANG
1
;
Wei PENG
2
;
Fang LIU
1
Author Information
1. Hunan University of Chinese Medicine， Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine， Changsha 410006， China
2. Institute of Oncology， Hunan Academy of Chinese Medicine， Changsha 410013， China
- Publication Type:Journal Article
- Keywords:
Jianpi Xiao'ai prescription;
colorectal cancer;
inducible nitric oxide synthase-arginase 1 （iNOS-ARG1）;
arginine metabolism;
mitochondrial dysfunction
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2026;32(13):99-111
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the mechanism by which Jianpi Xiao'ai prescription （JPXAP） inhibits colorectal cancer progression by regulating the inducible nitric oxide synthase-arginase 1 （iNOS-ARG1） metabolic axis and inducing mitochondrial reactive oxygen species （mito-ROS）-mediated mitochondrial structural and functional impairment. MethodsAn arginine metabolism disorder model of human colorectal cancer HCT116 cells was established by combined treatment with recombinant human interferon-γ （IFN-γ， 10 μg·L-1） and N（ω）-hydroxy-L-arginine （Nor-NOHA， 200 μmol·L-1） for 24 h， followed by intervention with 5%， 10%， or 20% JPXAP-containing serum. Cell proliferation was assessed using cell counting kit-8 （CCK-8）， 5-ethynyl-2′-deoxyuridine （EdU） staining， and colony formation assays. Cell invasion and migration were evaluated using Transwell chamber and wound healing assays. Mitochondrial membrane potential （MMP） and ROS levels were assessed by JC-1 and MitoSOX staining， respectively. Mitochondrial ultrastructure was observed by transmission electron microscopy （TEM）. The expression of iNOS， ARG1， and mitochondrial dynamics-related proteins， including mitofusin 2 （MFN2） and dynamin-related protein 1 （DRP1）， was analyzed by Western blot and immunofluorescence. The levels of L-arginine， citrulline， and urea were determined by colorimetric methods and enzyme-linked immunosorbent assay （ELISA）. ResultsCompared with the blank group， the model group exhibited significantly upregulated iNOS expression， downregulated ARG1 expression， a decreased ARG1/iNOS ratio， reduced L-arginine and urea levels， and increased citrulline levels （P<0.05）. Meanwhile， mito-ROS accumulation was significantly increased， the JC-1 red/green fluorescence ratio was decreased， and mitochondria showed swelling and cristae disruption， indicating that metabolic disorder induced mitochondrial injury. Compared with the model group， all JPXAP-treated groups further decreased the ARG1/iNOS ratio， enhanced nitric oxide （NO） and reactive nitrogen species accumulation， further reduced L-arginine and urea levels， and increased citrulline levels （P<0.01）. EdU-positive rate， colony formation rate， wound healing rate， and Transwell invasion number all decreased significantly with increasing serum concentration （P<0.01）. Mito-ROS levels were further elevated， and the JC-1 red/green ratio further decreased. TEM revealed aggravated mitochondrial swelling and vacuolization. MFN2 expression was downregulated and DRP1 expression was upregulated （P<0.01），in a dose-dependent manner. ConclusionJPXAP further activates NO-mediated oxidative/nitrosative stress under arginine metabolism imbalance， inducing mito-ROS accumulation， MMP collapse， and mitochondrial dynamics imbalance， thereby inhibiting colorectal cancer cell proliferation and migration. These findings reveal an antitumor mechanism of JPXAP based on coordinated targeting of the "metabolism-mitochondria" axis.
