Chinese Journal of Integrative Medicine最新文献

Objectives: To explore the effect of Pollen Pini (PP) on SMMC-7721 hepatoma cells.

Methods: The anti-proliferative effects of PP on SMMC-7721 cells were evaluated using cell counting kit-8 assay following 48 h treatment with concentrations ranging from 1.25 to 37.50 µg/µL Flow cytometry analysis at the half maximal inhibitory concentration (IC₅₀) revealed significant apoptosis induction and cell cycle alterations. For in vivo evaluation, SMMC-7721 xenograft-bearing nude mice were administered either vehicle (0.9% NaCl) or PP (500 mg/kg) via intraperitoneal injection every other day for 3 weeks. Subsequent tumor analysis included mass spectrometry-based proteomics and examination of phosphoinositide-3 kinase (PI3K)/protein kinase B (Akt)/Myc pathway proteins by immunohistochemistry and Western blot. Combination therapy with 25 µmol/L PI3K inhibitor and PP (IC₅₀) showed optimal efficacy, with Western blot revealing maximal protein modulation at 24 h.

Results: PP had an anti-tumor effect on SMMC-7721 cells in vitro, with the IC₅₀ concentration of 18.94 mg/mL. PP could promote the death of SMMC-7721 cells (P<0.01) and regulate the cell cycle more in G₀/G₁ phase cells (P<0.01). After the treatment effect of PP, the protein content of SMMC-7721 cells changed and the contents of cancer-promoting proteins PI3K, Akt and Myc decreased (P<0.01). PI3K inhibitor could reduce the proliferation of SMMC-7721 cells, and PI3K inhibitor combined with PP significantly reduced the expression of PI3K in SMMC-7721 cells (P<0.01).

Conclusion: PP has an anti-tumor effect on SMMC-7721 cells through the PI3K-Akt signaling pathway.

Objective: To investigate the anticancer activity of curzerene in colorectal cancer (CRC) in vitro and in vivo models.

Methods: HT29 and HCT8 cells were treated with different concentrations of curzerene (0, 20, 40, and 60 µg/mL) for 24 h. Cell viability was assessed using cell counting kit 8 assay, and cell proliferation was detected by colony-formation, then apoptosis rate was assessed by flow cytometry analysis. Mitochondrial membrane potential was measured using JC-1 assay kit. Intracellular calcium levels were examined using Fluo-3AM and Mag-fluo-3AM staining. Different inhibitors of cell death, including 3-methyladenine (3-MA), cloroquine (CQ), Nec-1, and carbobenzoxy-valyl-alanylaspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK), were also utilised to validate the death mechanisms. The binding ability of curzerene to mitogen-activated extracellular signal-regulated kinase (MEK) proteins was investigated by molecular docking. In addition, the expression of key proteins such as phosphated MEK (p-MEK), phosphated extracellular regulated protein kinase (p-ERK), B-cell lymphoma-2 (Bcl-2), Bcl associated X (Bax), poly ADP-ribose polymerase (PARP) and cleaved PARP were analysed by Western blot. Finally the viable HT29 cells were injected subcutaneously into the right dorsolateral abdomen of male BALB/c nude mice for in vivo potency assessment.

Results: Curzerene inhibited proliferation and induced apoptosis in HT29 and HCT8 cells in a time- and dose-dependent manner (all P<0.05). Subsequently, we demonstrated that the apoptosis inhibitor Z-VAD-FMK (P<0.05) but not 3-MA, CQ or necrostatin-1 rescued curzerene-induced cell death. Compared with the control group, 60 µg/mL curzerene increased the expression of cleaved PARP by affecting intracellular calcium distribution, reactive oxygen species (all P<0.01), decreasing mitochondrial membrane potential and the expressions of p-MEK, p-ERK, Bcl-2, and PARP (all P<0.05), and additionally increased the expression of cleaved PARP with a molecular binding energy of -7.1 kcal/mol. The results showed that curzerene treatment inhibited the activation of MEK/ERK signaling pathway, and pretreatment with the MEK activator C16-PAF significantly alleviated curzerene-induced cell death (all P<0.05). The results of in vivo experiments showed that curzerene significantly inhibited the growth of subcutaneous transplantation tumours in hormonal nude mice.

Conclusion: Curzerene induces apoptosis in CRC cells through inhibition of the MEK/ERK signaling pathway, which will hopefully be a potential chemotherapeutic agent for treating CRC patients.

Objective: To investigate the effect and mechanism of panaxadiol saponin component (PDS-C) in radiation-induced cardiac injury (RIHD).

Methods: BALB/c mice were radiated by X-ray to establish RIHD mouse models, and randomly divided into 3 groups by using a random number table method (n=10 in each group): the mice in the normal control and RIHD model groups received normal saline, and the PDS-C group was treated with PDS-C (80 mg/kg) for 12 weeks. Cardiac function was observed by ultrasound imaging; serum myocardial enzymes were tested; and pathological examination of cardiac tissue was performed. Differentially transcribed genes of cardiac tissue were analyzed between normal and RIHD mice. The expression levels of inflammatory factors and fibrotic proteins were detected by immunohistochemistry, immunohistofluorescence and Western blot, respectively. Irradiation H9c2 cells were incubated by PDS-C (20, 40, 80 mg/L) for detecting inflammatory factors and fibrotic proteins. Irradiation H9c2 cells were incubated by both phorbol 12-myristate 13-acetate (PMA) as nuclear factor κB (NF-κB) activator and BAY11-7082 as inhibitor to verify the effects of PDS-C on NF-κB transcription factor.

Results: Differentially expressed genes between normal and RIHD mice were involved mainly in the NF-κB transcription factor, which is related to cardiac inflammation. PDS-C effectively increased the cardiac ejection fraction and fractional shortening (P<0.05), but decreased the levels of the myocardial enzymes aspartate aminotransferase, creatine kinase, creatine kinase isoenzyme and lactate dehydrogenase in RIHD mice (P<0.05). Pathological examination of cardiac tissue revealed obvious edema, severe necrosis, and nuclear pyknosis in RIHD mice; however, PDS-C clearly alleviated the extent of cardiac damage. Furthermore, PDS-C downregulated interleukin (IL)-1, IL-6, Smad2, connective tissue growth factor, collagen II and NF-κB (P<0.05). Cytological experiments revealed that PDS-C decreased reactive oxygen species levels in irradiated H9C2 cells (P<0.05), and the expressions of the above inflammatory factors and fibrosis-related proteins were consistent with the results in mouse models.

Conclusion: PDS-C showed cardioprotective effect in alleviating inflammation and fibrosis through inhibition of the NF-κB signaling pathway.

Objective: To elucidate the effect of hydroxychavicol (HC) in combination with 5-fluorouracil (5-FU) on purine metabolism and apoptosis in colorectal cancer cell lines HT-29 and DLD-1.

Methods: The viability of HT-29 and DLD-1 cells when treated with HC, (0-1,000 µmol/L) 5-FU (0-100 µmol/L) alone, and HC+5-FU for 24 and 48 h was determined. Hypoxanthine (HPX) and xanthine oxidoreductase (XOR) were evaluated, as well as reactive oxygen species (ROS) levels and mitochondrial membrane potential (MMP). The expression levels of genes including nucleoside transporters equilibrative nucleotide transport 1 and 2 (ENT1 and ENT2), the proapoptotic gene Caspase-3 (CASP3), and the anti-apoptotic gene BCL2 were analysed by quantitative polymerase chain reaction.

Results: Both HPX and XOR levels in cells treated with HC+5-FU were significantly decreased (P<0.05) after 24 and 48 h compared to control cells. ROS levels in HT-29 and DLD-1 treated with HC+5-FU for 24 and 48 h were 26.2% and 21.4%, and 9.1% and 20.5%, respectively, significantly lower than control cells. MMP assays indicated mitochondrial depolarisation. In HT-29 cells, ENT1 and BCL2 were downregulated at 24 h, and CASP3 was upregulated at 48 h. In DLD-1 cells, ENT1 and ENT2 were downregulated, while CASP3 showed a transient decrease at 24 h.

Conclusions: The combination of HC + 5-FU demonstrated synergistic effects in HT-29 and DLD-1 cells, disrupting oxidative balance and purine metabolism, as reflected in reduced hypoxanthine levels, XOR activity, and ROS production. This treatment also induced mitochondrial membrane depolarisation and altered apoptosis-related gene expression, supporting its role in apoptosis induction.