ABSTRACT
OBJECTIVE: To investigate the effects of interference of riboflavin (RIB) metabolic pathways on adhesion and migration to cisplatin (DDP) in ovarian cancer HO8910 cells. METHODS: Intervention RIB metabolic pathways by lentiviral vector harboring shRNA of riboflavin transporter 2 (RFT2) and chlorpromazine (CHL), a competitive inhibitor of RIB. HO8910 ovarian cancer cell line was divided into normal control group, shRNA control group, RFT2 shRNA group, CHL (50 μmol·L-1) group, DDP (20 μmol·L-1) group, RFT2 shRNA+DDP group, CHL + DDP group and DDP + RIB (20 μmol·L-1) group. Each group cells were collected after treatment 48 h according to the design. And then cell adhesive abilities were detected by adhesion experiment, the cells invasive abilities were observed by transwell method, the protein expressions of VEGF, MMP9 and MMP2 were detected by Western blot, and the expressions of TNF-α, NF-κB/p65 were assayed with laser confocal microscopy. RESULTS: Compared with the sole DDP treatment group, RFT2 shRNA or CHL combination with DDP had great advantages in reducing cell adhesion and migration viabilities, deceasing expressions of MM9 and MMP2, reducing cell expressions of TNF-α and NF-κB/p65. However, the RIB could weaken the effects of DDP on HO8910 cell. CONCLUSION: Interference metabolic pathway of RIB can enhance DDP effects on adhesion and migration viabilities of ovarian cancer HO8910 cell lines, and the effects are associated with blocking the pathway of TNF-α/NF-κB; However, RIB could attenuate the anti-tumor effects of cisplatin on HO8910 cell.
ABSTRACT
OBJECTIVE: To investigate the effects of icariin (ICA) on autophagy and activation of HSC-T6 cell line induced by lipopolysaccharide (LPS), and further research whether this effect is associated with FXR protein. METHODS: The HSC-T6 hepatic stellate cell was stimulated with LPS and then treated with ICA of three different doses and TLR4 siRNA. Autophagy changes were observed through the acridine orange (AO) and MDC staining, expression of Beclin-1 and LC3B were determined via laser scanning confocal microscopy, cell mitochondrial membrane potentials were detected using flow cytometry and the proteins of NF-κB (p65), LC3B-II, CollagenI, TGF-(β1, α-SMA were measured through Western blot. Furthermore, the ICA treatment group was intervened with Z-Gug-gulsterone(Z-Gug). The expressions of LC3B, Beclin-1 and FXR were measured through Western blot. RESULTS: Compared with the normal group, LPS significantly increased autophagy, up-regulated expression of Beclin-1 and LC3B, decreased mitochondrial membrane protentials and increased expression of NF-κB(p65), LC3B-II, Collagen I, TGF-β1 and α-SMA. However, the effects of LPS on HSC-T6 could be partly inhibited by TLR4 siRNA and ICA in a dose-dependent manner. Moreover, ICA increased expression of FXR and could be blocked by Z-Gug treatment. CONCLUSION: ICA ameliorated LPS/TLR4 can induce hepatic stellate cell activation by reduction autophagy and association with FXR protein.
ABSTRACT
BACKGROUND: Acute pulmonary embolism (APE) is a disorder involving the pulmonary circulation resulting from a blockage of the pulmonary artery. The present study aimed to investigate the effects of aspirin on the nuclear factor-κB (NF-κB) activity in a rat model of APE. METHODS: A total of 108 healthy male Sprague-Dawley rats were randomly assigned into six groups (n=18 rats per group): control group, sham operation group, APE model group, and low-, medium- and high-dose aspirin groups. Six, 24, and 72 hours after the induction of APE, rats in the low-, medium- and high-dose aspirin groups were given aspirin at a respective daily dose of 150, 300, and 600 mg/kg by gavage for three consecutive days. Rats in the other groups were treated with equal volumes of normal saline. Six rats in each group were anesthetized with 10％ chloral hydrate solution at each time point, and then the lung tissues were colected and analyzed using immunohistochemical staining. RESULTS: Positive immunohistochemical staining was present in the bronchial epithelial cells, alveolar cells, macrophages, and surrounding bronchial smooth muscle cells. When compared with the APE model group, the number of positive cells was significantly lower in the other groups at each time point (P<0.001). Statistically significant differences were also observed among the aspirin-treated groups at 6 hours (P<0.05,P<0.001). Compared with the APE model group, NF-κB protein expression was reduced in the other groups at each time point (P<0.05,P<0.001). Rats from the APE model group had thrombosis, damaged alveolar walls, and pulmonary hemorrhage, along with different degrees of infl ammatory cellular infiltration at each time point. However, pathological changes such as pulmonary hemorrhage and infiltration of inflammatory cells were attenuated after the aspirin treatment. CONCLUSION: Aspirin can significantly inhibit NF-κB activity in the lung of rats with APE in a dose-dependent manner, and can alleviate lung injury after APE.