1.Mechanistic investigation of FOXO4 in Celastrus orbiculatus extract-mediated inhibition of aerobic glycolysis in the therapy of gastric precancerous lesions
Ziwei PAN ; Junsong WEN ; Xuedan YAO ; Yanqing LIU ; Yaodong ZHU
Acta Universitatis Medicinalis Anhui 2026;61(3):462-469
ObjectiveTo investigate the effects of forkhead box protein O4 (FOXO4) expression on gastric precancerous lesions (GPL), and to clarify its mechanism in mediating the therapeutic effect of Celastrus orbiculatus extract (COE) on GPL by regulating aerobic glycolysis. MethodsReferring to the previously established combined modeling protocol in our research group, a rat model of gastric precancerous lesions (GPL) was constructed through the following procedures: rats were given free access to 170 μg/mL N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) solution for ad libitum drinking, fed with a diet supplemented with 0.03% ranitidine hydrochloride, and treated with a cycling regimen of “2-day feeding followed by 1-day fasting”. Specifically, on the afternoon of each fasting day, the rats received intragastric administration of 2% sodium salicylate at a dose of 10 mL/kg body weight. FOXO4-silenced and overexpression model rats were constructed by tail vein injection of plasmids. The rats were randomly divided into control, model, COE, overexpression negative control (OE-NC), FOXO4 overexpression (OE-FOXO4), OE-FOXO4+COE, silencing negative control (shNC), FOXO4 silencing (shFOXO4) and shFOXO4+COE groups. Gastric mucosal histopathological changes were observed in each group. Lactic acid content in gastric mucosal tissues was detected by colorimetry. The expression levels of FOXO4, HK2, PKM2, LDHA and GLUT1 were evaluated by immunohistochemistry (IHC), and their mRNA levels were determined by RT-PCR.Results Compared with the control group, the COE, OE-FOXO4 and OE-FOXO4+COE groups exhibited significantly improved gastric mucosal lesions, reduced lactic acid levels, weakened expression of aerobic glycolysis-related proteins (PKM2, HK2, LDHA, GLUT1), and enhanced FOXO4 expression. The OE-FOXO4+COE group showed the lowest lactic acid level and more pronounced changes in related protein expression compared with the COE and OE-FOXO4 groups. In contrast, the shFOXO4 and shFOXO4+COE groups displayed increased lactic acid levels, enhanced expression of aerobic glycolysis-related proteins, and reduced FOXO4 expression compared with the model group.ConclusionFOXO4 expression is involved in the inhibitory effect of COE on GPL, possibly by regulating the aerobic glycolysis process.
3.UPLC-Q-TOF-MS Analysis on Chemical Constituents of Classical Prescription Xiehuang San Standard Decoction
Wan XIAO ; Siqi WANG ; Jiazheng LI ; Xuedan FU ; Jianming JU ; An KANG ; Weifeng YAO ; Hailang JIANG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(6):569-581
OBJECTIVE To analyze the chemical constituents from classical prescription Xiehuang San(XHS)standard decoc-tion by UPLC-Q-TOF-MS technology,and classify the chemical composition and analyze the representative components.METHODS Acquity HSS T3 column(2.1 mm×100 mm,1.8 μm)was used as the chromatographic column,with 0.1%formic acid solution-0.1%formic acid acetonitrile as the mobile phase for gradient elution.The volume flow rate was 0.4 mL·min-1 and the column tem-perature was 40℃.Mass spectrometry data of XHS were collected in positive and negative ion modes.The chemical constituents from classical prescription XHS were analyzed and identified by Masslynx 4.1 software comparison with reference materials,mass spectrome-try data analysis and reference to relevant literature.RESULTS A total of 107 compounds were analyzed and identified from XHS,including 45 flavonoids,27 triterpenoids,11 monoterpenoids,10 phenylpropanoids,6 chromogenic ketones,5 alkaloids and 3 other other compounds.CONCLUSION The study provides an experimental basis for the further research on the substance basis and qual-ity control of XHS.

Result Analysis
Print
Save
E-mail