Ophiopogon Polysaccharide Liposome Regulated the Immune Activity of Kupffer Cell through miR-4796.

The purpose of this article is to study the effects and mechanism of miR-4796 in the process of ophiopogon polysaccharide liposome (OPL) regulation of the immune activity of Kupffer cells (KCs). In this study, KCs were used as cell models, and were treated with OPL in different concentrations after being transfected with miR-4796 mimic or miR-4796 inhibitor. Firstly, the secretion of NO and iNOS, phagocytic activity, the expression of surface molecules CD14 and MHC II, apoptosis and ROS secretion were measured by Griess, flow cytometry, fluorescence staining and ELISA. Then, real-time PCR and Western blot were used to measure the expression of TLR4, IKKß, MyD88 and NF-κB in the TLR4-NF-κB signaling pathway. The results showed that after transfection with miR-4796 mimic, the secretion of NO and iNOS, cell migration, cell phagocytosis and expression levels of CD14 and MHC II in the OPL group were significantly higher than those in the miR-4796 mimic control group (p < 0.05; p < 0.01). In addition, the mRNA and protein expression levels of TLR4, MyD88 and NF-κB were significantly higher than those in miR-4796 mimic control group (p < 0.05; p < 0.01). After transfection with miR-4796 inhibitor, the secretion of NO and iNOS, cell migration, cell phagocytosis, expression of CD14 and MHCII in OPL group were significantly higher than those in the miR-4796 inhibitor control group (p < 0.05; p < 0.01). These results indicated that OPL could regulate the immune activity of KCs by regulating miR-4796 and activating the TLR4-NF-κB signaling pathway.

The effect of miR-1338 on the immunomodulatory activity of ophiopogon polysaccharide liposome.

This study is to investigate the effect of microRNA-1338 (miR-1338) on the activity of Kupffer cells (KCs) and its mechanism regulated by ophiopogon polysaccharide liposome (OPL). KCs was treated with different OPL after transfected with miR-1338 mimic and miR-1338 inhibitor. The secretion of NO and iNOS, the expression of catalase (CAT) and peroxidase (POD), the phagocytic activity, the expression of CD14 and MHC II, the apoptosis and the secretion of ROS were measured. In addition, the expressions of key signal factors TLR4, IKKß, MyD88 and NF-κB in NF-κB signaling pathway were measured by real-time PCR and Western blot (WB). The results showed that OPL could promote the secretion of iNOS, the expression of POD, the phagocytosis, the mRNA expression of TLR4, MyD88, IKKß and NF-κB, the protein expression of TLR4 and NF-κB, and inhibit the cell apoptosis and ROS secretion after transfected with miR-1338 mimic. After transfected with miR-1338 inhibitor, OPL could promote the secretion of NO and iNOS, the expression of POD, cell migration, phagocytosis, and inhibit cell apoptosis. Meanwhile, the mRNA expression of TLR4, MyD88, IKKß and NF-κB and the protein expression of TLR4, MyD88 and NF-κB were promoted. These results suggested that OPL could activate TLR4-NF-κB signaling pathway and thereby improve the activity of KCs by regulating miR-1338.

Activation of macrophages by the ophiopogon polysaccharide liposome from the root tuber of Ophiopogon japonicus.

This study evaluated the immunomodulatory effects of ophiopogon polysaccharide liposome (OPL) on macrophages in vitro. The phagocytic activity, the secretion of nitric oxide (NO) and inducible nitric oxide synthase (iNOS), the level of cytokines, chemokines and the expression of CD14 and MHC-II costimulatory molecules were measured. Results showed that OPL could significantly improve the phagocytic activity and the level of IL-1ß, TNF-α, MCP-1 and MIP-1ß, promote the secretion of NO and iNOS, and enhance the expression of CD14 and MHC-II costimulatory molecules in the peritoneal macrophages of mice compared with ophiopogon polysaccharide (OP). Altogether, these results suggested that OPL could activate macrophages, and the efficacy was significantly superior to OP. Therefore, OPL would be exploited in a potent immunomodulators. Moreover, it also provided the theoretical basis for further studying the mechanism of OPL on improving the immune response.

Antioxidative and immunological activities of ophiopogon polysaccharide liposome from the root of Ophiopogon japonicus.

The antioxidative and immunoregulatory activities of ophiopogon polysaccharide liposome (OPL) were investigated. The results demonstrated that the radical-scavenging activity increased along with the increasing of OPL concentrations, and the scavenging capability of OPL was slightly stronger than ophiopogon polysaccharide (OP) in vitro. In vivo, OPL at high and medium doses could significantly improve the levels of SOD and GSH-Px, and decrease the levels of MPO, XOD and MDA compared with OP. In addition, OPL could not only significantly enhance the splenocyte proliferation and the proportion of CD4(+) and CD8(+) T cells, but also improve the levels of cytokines, antigen-specific antibody titers and immune organ index in the mice immunized with OVA compared with OP. These results indicated that the antioxidative and immunoregulatory activities of OP were significantly enhanced after encapsulated with liposome. Therefore, OPL would be expected to exploit into a new-type of functional food as both medicine and food.

The adjuvanticity of ophiopogon polysaccharide liposome against an inactivated porcine parvovirus vaccine in mice.

In this study, the adjuvant activity of ophiopogon polysaccharide liposome (OPL) was investigated. The effects of OPL on the splenic lymphocyte proliferation of mice were measured in vitro. The results showed that OPL could significantly promote lymphocyte proliferation singly or synergistically with PHA and LPS and that the effect was better than ophiopogon polysaccharide (OP) at most of concentrations. The adjuvant activities of OPL, OP and mineral oil were compared in BALB/c mice inoculated with inactivated PPV in vivo. The results showed that OPL could significantly enhance lymphocyte proliferation, increase the proportion of CD4(+) and CD8(+) T cells, improve the HI antibody titre and specific IgG response, and promote the production of cytokines, and the efficacy of OPL was significantly better than that of OP. In addition, OPL significantly improved the cellular immune response compared with oil adjuvant. These results suggested that OPL possess superior adjuvanticity and that a medium dose had the best efficacy. Therefore, OPL can be used as an effective immune adjuvant for an inactivated PPV vaccine.

Ophiopogon polysaccharide liposome can enhance the non-specific and specific immune response in chickens.

The purpose of this study is to investigate the immune-enhancing activity of ophiopogon polysaccharide liposome (OPL). In non-specific immune response experiment, the phagocytosis and cytokines secretion of peritoneal macrophages in vitro and in vivo were performed. In specific immune response experiment, the activity of OPL was measured on chickens which were vaccinated with Newcastle disease (ND) vaccine and then challenged with ND virus at 49-old-day. The results showed that OPL could significantly promote the phagocytosis of macrophages and induce the secretion of IL-2 and IL-6 in vitro; OPL at high and medium doses could significantly improve the phagocytosic index, promote lymphocyte proliferation, increase the proportion of T lymphocyte subpopulations (CD4(+) and CD8(+)), enhance antibody titer and improve the protective rate in vivo. Moreover, its efficacy was significantly better than ophiopogon polysaccharide (OP). These results indicated that the immune-enhancing activity of OP was significantly improved after encapsulated with liposome.

Preparation and optimization of ophiopogon polysaccharide liposome and its activity on Kupffer cells.

The purpose of this study was to prepare and optimize ophiopogon polysaccharide liposome (OPL), and to improve the immune-enhancing activity of ophiopogon polysaccharide (OP). OPL was prepared and optimized using the methods of reverse-phase evaporation and response surface methodology. The property was evaluated with particle size, zeta potential, and morphology. The results showed that the optimum preparation conditions were: soybean phosphatide to OP ratio of 9.5:1, soybean phospholipid to cholesterol ratio of 8:1, and chloroform to phosphate-buffered saline ratio of 3:1. Subsequently, the immune-enhancing activity of OPL on Kupffer cells (KCs) was performed. The results showed that OPL could significantly promote the phagocytosis of KCs, induce the secretion of nitric oxide, induced nitric oxide synthase, IL-6 and IL-12, and improve the expression of CD80 and CD86 compared with OP at 125-7.813 µg mL(-1). These results indicated that the immune-enhancing activity of OP was significantly improved after encapsulated with liposome. Therefore, liposome would be expected to exploit into a new-type preparation of OP.