ABSTRACT
In this study, an immunostimulating particulate β-glucan was isolated from a hot alkaline extract of the fruiting bodies of Ganoderma lucidum. The optimum conditions of 8 hours treatment time, 1∶20 solid - liquid ratio and 55 ℃ for the alkaline extract process were obtained after investigating by single-factor experiments and Box-Benhnken design in terms of the Ganoderma lucidum particulate β-glucan (GLG) increment, and these conditions resulted in a GLG yield of 8.57%. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Jiangsu Provincial Academy of Chinese Medicine. The result showed that resident macrophages were effectively activated by GLG, such as with the up-regulation of co-stimulatory molecules, the secretion of cytokines and phagocytic uptake. GLG could also promote the proliferation of spleen lymphocytes in mice. In addition, IFN-γ production of spleen CD4+ T cells and cytotoxic T lymphocyte (CTL) responses were significantly enhanced on GLG orally treatment, which ultimately resulted in significantly decreased tumor burden. Taken together, these data suggest that GLG might act as an immune stimulator to exert antitumor effects.
ABSTRACT
Objective Solamargine (SM), with its anti-inflammatory and anti-tumor effects, inhibits the proliferation and promotes the apoptosis of various tumor cells. This study was to investigate the effects of SM on the proliferation and apoptosis of human esophageal cancer KYSE150 cells and its action mechanism. Methods We treated KYSE150 cells with SM at the concentrations of 0 (the blank control group), 2, 4, 6 and 8 μmol/L for 24 hours. Then, we observed the morphological changes of the cells under the inverted microscope, detected their proliferation and apoptosis by MTT assay and flow cytometry respectively, and determined the expressions of the classical NF-κB signaling pathway-related proteins NF-κB, p-NF-κB, IKKα, IKKβ, IkBα and p-IkBα) and apoptosis-related proteins Bax, caspase-3, cleaved caspase-3 and Bcl-2 in different groups of the cells by Western blot. Results Compared with the blank control, the inhibition rate of the proliferation of the KYSE150 cells in the 2, 4, 6 and 8 μmol/L SM groups was increased significantly in a concentration-dependent manner (0 vs [15.03 ± 0.15]%, [47.94 ± 1.74]%, [68.72 ± 0.47]% and [77.51 ± 1.70]%, P<0.05), and so was the apoptosis rate ([8.17 ± 0.51]% vs [14.50 ± 0.73]%, [18.57 ± 2.08]%, [65.10 ± 10.88]% and [81.55 ± 5.48]%, P<0.05). The expression of the apoptosis-related protein Bax in the SM treated cells was up-regulated, those of Bcl-2, IKKα, IKKβ and p-IkBα down-regulated, and the activity of caspase-3 and cleaved caspase-3 promoted, all in a concentration-dependent manner, with statistically significant differences between the blank control and the 4, 6 and 8 μmol/L SM groups (P<0.05). Statistically significant differences were also found in the expressions of NF-κB, p-NF-κB and IkBα between the blank control and the 6 and 8 μmol/L SM groups (P<0.05). Conclusion Solamargine can significantly inhibit the proliferation and promote the apoptosis of KYSE150 cells, probably by suppressing the classical NF-κB signaling pathway.
ABSTRACT
Objective The metastasis mechanism of cholangiocarcinoma is complex, which may be related to epithelial-mesenchymal transition(EMT). This study focused on investigating the inhibition effects of allicin on TGF-β1 induced epithelium mesenchymal transition of human cholangiocarcinoma cells and its related mechanism, and providing theoretical basis for the application of allicin in the treatment of cholangiocarcinoma. Methods MTT assay were used to detect the inhibition effects of different concentrations of allicin on the human cholangiocarcinoma RBE cell proliferation, and the drug concentration of allicin was determined by IC50 of 24 h. The RBE cells were cultured and divided into control group, allicin group(130.7μmol/L), TGF-β1 group(10ng/mL) and allicin+ TGF-β1 group(130.7μmol/L+10ng/mL). Wound scratch and transwell invasion assay were performed to detect the migration and invasion ability of RBE cells after 24 hours. Western blots were applied to detect expression of EMT-related proteins (E-Cadherin, N-Cadherin, Vimentin, Snail) and NF-κB signaling pathways. Results The migration rates in allicin group and allicin+ TGF-β1 group were both decreased compared with that in the control group ( 9.25% ± 0.36% vs 28.19 %±0.66%, P<0.05) and TGF⁃β1 group(13.91%±0.75% vs 49.22%±0.27%, P<0.05). The invasion rates in allicin group and allicin+ TGF-β1 group were also decreased compared with that in the control group (6.59%±0.06% vs 33.48%±0.04%, P<0.05) and TGF⁃β1 group(9.4%± 0.05% vs 40.21%±0.12%, P<0.05). Compared with the control group, E-Cadherin expression was significantly increased, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly decreased in the allicin group (P<0.05). Compared with TGF-β1 group, E-Cadherin expression was significantly up-regulated, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly down-regulated in the allicin+ TGF-β1 group (P<0.05)． Conclusion These results indicate that allicin can inhibit the EMT induced by TGF-β1 on the human cholangiocarcinoma cell by blocking NF-κB signaling pathway, which may have potential value to be the drug candidate for the treatment of human cholangiocarcinoma in future.
ABSTRACT
The aim of this study was to investigate the mechanisms of mono-functional alkylating agent MNNG to damage human gastric epithelial GES-1 cells and roles of Wnt/β-catenin signaling pathway in the process. The GES-1 cells were treated with MNNG (2 × 10 mol/L) for 24 h. The morphological changes of the GES-1 cells were observed under inverted microscope 2 d after treatment. The cell viability was measured by MTT assay. The apoptosis and cell cycle distribution of the GES-1 cells were analyzed by flow cytometry. The mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7 in the GES-1 cells were detected by qPCR. The protein expressions of β-catenin, GSK-3β, p-GSK-3β and c-Met were determined by Western blot. The results showed that MNNG induced the injury of GES-1 cells and changed the normal cell morphology to irregular long spindle shape. MNNG induced the apoptosis of GES-1 cells and blocked the cell cycle progression obviously. MNNG up-regulated the mRNA expressions of β-catenin, GSK-3β, c-Met and MMP7, and increased the protein expressions of β-catenin, GSK-3β and p-GSK-3β. These results suggest that the damage of GES-1 cells induced by MNNG may be related to the activation of Wnt/β-catenin signaling pathway, which will provide the basis for the study of cell model of gastric mucosal cell injury.
ABSTRACT
Objective: To study the possible mechanisms of Chinese materia medica Kangmin Pingchuan Formula via regulating the balance of regulatory T cells (Tregs) and Th17 cells. Methods: The female BALB/c mice were divided into five groups: control, model, Prednisone (5.5 mg/kg), Kangmin Pingchuan Formula low- (4.95 g/kg) and high-dose (9.90 g/kg) groups; Ovalbumin was used to make the asthmatic model of mice, the drug was ig administered daily after atomization for consecutive 15 d, then the mice were killed after the last administration. The paraffin-embedded tissue sections of the lungs in mice were stained by HE; Th17 cells and Tregs in blood were detected by flow cytometry; IL-4, TGF-β, and TNF-α in serum of mice were detected by ELISA assay. Results: HE staining showed that Kangmin Pingchuan Formula could alleviate the inflammation in the lung tissue of mice, decrease the proportion of Th17 cells, increase the proportion of Tregs in blood, promote TGF-β levels, and inhibit TNF-α levels in blood. Conclusion: Kangmin Pingchuan Formula could treat the asthma through impacting the proportion of Th17 cells and Tregs in blood as well as the levels of related inflammatory cytokines in mice.