Protective effect of Pai-Nong-San against AOM/DSS-induced CAC in mice through inhibiting the Wnt signaling pathway.

Pai-Nong-San (PNS), a prescription of traditional Chinese medicine, has been used for years to treat abscessation-induced diseases including colitis and colorectal cancer. This study was aimed to investigate the preventive effects and possible protective mechanism of PNS on a colitis-associated colorectal cancer (CAC) mouse model induced by azoxymethane (AOM)/dextran sodium sulfate (DSS). The macroscopic and histopathologic examinations of colon injury and DAI score were observed. The inflammatory indicators of intestinal immunity were determined by immunohistochemistry and immunofluorescence. The high throughput 16S rRNA sequence of gut microbiota in the feces of mice was performed. Western blot was used to investigate the protein expression of the Wnt signaling pathway in colon tissues. PNS improved colon injury, as manifested by the alleviation of hematochezia, decreased DAI score, increased colon length, and reversal of pathological changes. PNS treatment protected against AOM/DSS-induced colon inflammation by regulating the expression of CD4+ and CD8+ T cells, inhibiting the production of HIF-α, IL-6, and TNF-α, and promoting the expression of IL-4 and IFN-γ in colon tissues. Meanwhile, PNS improved the components of gut microbiota, as measured by the adjusted levels of Firmicutes, Bacteroidetes, Proteobacteria, and Lactobacillus. PNS down-regulated the protein expression of p-GSK-3ß, ß-catenin, and c-Myc, while up-regulating the GSK-3ß and p-ß-catenin in colon tissues of CAC mice. In conclusion, our results suggested that PNS exhibits protective effect on AOM/DSS-induced colon injury and alleviates the development of CAC through suppressing inflammation, improving gut microbiota, and inhibiting the Wnt signaling pathway.

[Effect of supplementing Qi and activating blood circulation method on protein uptake in NRK-52E cells injured by high glucose].

OBJECTIVE: To study the effect of supplementing Qi and activating blood circulation method with danshen and huangqi on protein uptake in NRK-52E cells injured by high glucose. METHODS: The influence of combined use of huangqi and danshen on NRK-52E cells proliferation were determined by MTT method, FITC-BSA uptake were observed by fluorescence microscope and analyzed by fluorescence spectrophotometer, and megalin expression were detected by the method of immunohistochemistry. RESULTS: Combined use of huangqi and danshen could strengthen its protection effect. The FITC-BSA uptake and expression of megalin in NRK-52E cells were decreased in high glucose environment. Combined use of huangqi and danshen could increase FITC-BSA uptake and megalin expression in high glucose-injured NRK-52E cells. CONCLUSION: The method of supplementing Qi and activating blood circulation can protect high glucose injured NRK-52E cells and improve the function of protein uptake by increasing expression of megalin.

Layer-by-layer assembly of chitosan stabilized multilayered liposomes for paclitaxel delivery.

Paclitaxel (PTX) loaded multilayered liposomes were prepared using layer-by-layer assembly in an effort to improve the stabilization of the liposomal compositions for PTX delivery. Stearyl amine was used to provide positive charge to the PTX-liposomes, and subsequently coated with anionic polyacrylic acid (PAA) followed by cationic chitosan. Various process variables were optimized and the optimum formulation was found to have particle size of 215 ± 17 nm, zeta potential of +27.9 ± 3.4 mV and encapsulation efficiency of 70.93 ± 2.39%. The lyophilized chitosan-PAA-PTX-liposomes formulation was stable in simulated gastrointestinal fluids and at different environmental conditions (4 °C and 25 °C). In vitro drug release experiments demonstrated that chitosan-PAA-PTX-liposomes formulation exhibited obvious sustained release behaviors compared to PTX-liposomes. Furthermore, chitosan-PAA-PTX-liposomes formulation revealed enhanced PTX induced cytotoxicity in human cervical cancer cell culture experiments compared to PTX-liposomes. In conclusion, the approach presented herein will provide a promising solution for PTX delivery.

Macrophage receptors of polysaccharide isolated from a marine filamentous fungus Phoma herbarum YS4108.

AIM: YCP, a novel (1,4)-alpha-D-glucan, was isolated from the mycelium of the marine filamentous fungus Phoma herbarum YS4108. In this work, we investigated a YCP-binding cellular receptor expressed by macrophages and the intracellular signal transduction pathways involved in YCP-induced macrophage activation. METHODS: Fluorescence-labeled YCP (fl-YCP) was prepared using the CDAP-activation method. Fluorescence confocal laser microscopy and fluorescence-activated cell sorting (FACS) were used to analyze the effect of fl-YCP on macrophages. To characterize the properties of the YCP receptor, carbohydrates and antibodies were used to inhibit the binding of fl-YCP to macrophages. Moreover, we investigated the role of membrane receptors Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), Toll-like receptor 6 (TLR6) and complement receptor 3 (CR3). We also examined the role of the p38 kinase pathway in mediating nitric oxide (NO) production. RESULTS: YCP had an in vitro stimulatory effect on the release of NO in macrophage, and fl-YCP can bind directly to receptors on the surface of macrophages in a time- and dose-dependent manner. Competition studies show that LPS, laminarin, anti-TLR4 antibody and anti-CD11b (CR3) antibody could inhibit fl-YCP binding to macrophages. Conversely, mannose, anti-TLR2 and anti-TLR6 antibody could not. Treatment of RAW264.7 cells with YCP resulted in significant activation of p38 in a time-dependent manner. The specific p38 inhibitor SB203580 abrogated YCP-induced NO generation. Treatment of RAW264.7 cells with anti-TLR4 antibody and anti-CR3 antibody significantly reduced YCP-induced NO production and p38 activation. CONCLUSION: We have demonstrated that YCP-induced NO production occurs through the TLR4 and CR3 membrane receptors in a p38 kinase-dependent manner in macrophages.Acta Pharmacologica Sinica (2009) 30: 1008-1014; doi: 10.1038/aps.2009.93.