Analysis of enhanced efficacy mechanism of Terminalia chebula sand stir-frying processing on ulcerative colitis based on Nrf2/HO-1/GPX4 signaling pathway and metabolomics
Objective To explore the mechanism of Terminalia chebula before and after sand stir-frying processing on ulcerative colitis(UC)based on nuclear factor E2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/glutathione peroxidase 4(GPX4)signaling pathway and serum metabolomics,and to reveal the enhanced efficacy mechanism of T.chebula processing.Methods UC rat model was established using the dextran sulfate sodium(DSS)method.The raw and sand stir-fried T.chebula were administered by gavage to observe the general signs,disease activity index(DAI),the length of colorectum,and colon pathological status in rats.The mRNA relative expression levels of tumor necrosis factor-alpha(TNF-α),interleukin-1β(IL-1β),IL-10,Nrf2,HO-1,GPX4,and superoxide dismutase(SOD)in colon tissues were detected by quantitative real-time polymerase chain reaction(qRT-PCR).The protein relative expression levels of Nrf2,HO-1,and GPX4 in colon tissues were detected by western blotting.Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)was performed to detect the serum non-targeted metabolites in rats,screen for potential biomarkers in serum,and analyze potential metabolic pathways in combination with the human metabolome database(HMDB)and Kyoto encyclopedia of genes and genomes(KEGG).Results The raw and sand stir-fried T.chebula both mitigated the disease symptoms of DSS-induced UC rats,reduced the DAI scores,restored the length of colorectum,and improved colonic pathological damage.Compared to the model group,the mRNA expression levels of inflammatory cytokines and oxidative stress cytokines in the colon of rats were significantly reversed(P＜0.05,0.01,0.001).The protein expression of Nrf2,HO-1,and GPX4 was restored,with all factors except for the regulation of GPX4 by the raw T.chebula group demonstrating significant effects(P＜0.05,0.01,0.001).Compared to the raw T.chebula group,the sand stir-fried T.chebula demonstrated superior efficacy in the above effects.Among these effects,the sand stir-fried T.chebula significantly regulated the expression of Nrf2 and HO-1 mRNA,as well as HO-1 protein expression(P＜0.001).A total of 36 potential biomarkers were screened,with the raw and sand stir-fried T.chebula regulating 10 and 12 potential biomarkers,respectively.The sand stir-fried T.chebula affected eight signaling pathways,including arginine biosynthesis,to restore the balance of metabolites in the body.Conclusion After being processed by sand stir-frying,T.chebula may exert a stronger ameliorative effect on UC by inhibiting inflammatory responses,interfering with the Nrf2/HO-1/GPX4 signaling pathway,and regulating metabolic pathways such as serum metabolites and arginine biosynthesis.
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