Psoralen and isopsoralen improve lipid metabolism disorder via inhibition of NF-κB activation in LO2 cells / 中国中药杂志

The aim of this paper was to investigate the mechanism and effect of psoralen and isopsoralen in the treatment of lipid accumulation in LO2 cells. Human LO2 cells nonalcoholic fatty liver models were established by using palmitic acid( PA). Then psoralen and isopsoralen were administered for intervention. Intracellular triglyceride( TG) and total cholesterol( TC) content,the cell supernatant alanine aminotransferase( ALT) and aspartate aminotransferase( AST) levels were determined by enzyme method. Cell supernatant proinflammatory cytokines( IL-6,TNF-α) and chemokines( IL-8,MCP-1) were determined by ELISA method. Western blot method was conducted to detect the protein expression of intracellular nuclear factor( NF-κB) p65 phosphorylation( p-p65),nonphosphorylated protein( p65),and transforming factor TGF-β1. Result showed that as compared with the model group,intracellular TG and TC levels,the cell supernatant ALT and AST levels,proinflammatory cytokines and chemokines were decreased( P < 0. 01,P <0. 05); the p-p65/p65 ratio and TGF-β1 protein expression were also significantly decreased( P< 0. 01,P< 0. 05) in psoralen intervention group. As compared with the model cells,intracellular TG content had no significant changes,but all the other indexes were reduced( P<0. 01,P<0. 05) in the cells of isopsoralen intervention group. Psoralen exhibited better effect than isopsoralen( P< 0. 01,P<0. 05). It is concluded that psoralen could improve the adipogenesis of LO2 cells induced by PA; both psoralen and isopsoralen are effective in ameliorating LO2 cells injury induced by PA,reducing inflammation via inhibiting the activation of NF-κB and down-regulating the expression of TGF-β1.

Toxicity testing of four silver nanoparticle-coated dental castings in 3-D LO2 cell cultures / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

To address the controversial issue of the toxicity of dental alloys and silver nanoparticles in medical applications, an in vivo-like LO2 3-D model was constructed within polyvinylidene fluoride hollow fiber materials to mimic the microenvironment of liver tissue. The use of microscopy methods and the measurement of liver-specific functions optimized the model for best cell performances and also proved the superiority of the 3-D LO2 model when compared with the traditional monolayer model. Toxicity tests were conducted using the newly constructed model, finding that four dental castings coated with silver nanoparticles were toxic to human hepatocytes after cell viability assays. In general, the toxicity of both the castings and the coated silver nanoparticles aggravated as time increased, yet the nanoparticles attenuated the general toxicity by preventing metal ion release, especially at high concentrations.

Effects of three Wenyang Jianpi Tang on cell proliferation and apoptosis of nonalcoholic fatty liver cells / 中国中药杂志

To investigate the effects of Sijunzi Tang, Lizhong Tang and Fuzi Lizhong Tang on the cell proliferation and apoptosis of nonalcoholic fatty liver cells through the nonalcoholic fatty liver cell model established by inducing L02 cells with oleic acid. Different concentrations of oleic acid were added into L02 cells to induce the nonalcoholic fatty liver cell model. Oil red O staining was used to observe fatty droplets of fatty liver cells. Automatic biochemical analyzer was used to detect the levels of aspartic transaminase(AST), alanine aminotransferase(ALT), total cholesterol(TC), and triglyceride(TG) in the cell supernatants. There were five groups, namely normal group, model group, model and Sijunzi Tang group, model and Lizhong Tang group, and model and Fuzi Lizhong Tang group. The cell proliferation and apoptosis of the five groups were detected by MTT colorimetry test and flow cytometer. The expressions of PCNA, cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, Bax and Bcl-2 proteins of the five groups were detected by Western blot. The oil red O staining results showed that the optimum concentration of oleic acid that was used to induce nonalcoholic fatty liver cell models was 80 mg•L-1. The levels of AST, ALT, TC and TG in the nonalcoholic fatty liver cell supernatants were higher than that in normal liver cell supernatants(P<0.01). MTT colorimetry test and flow cytometer results showed that all of Sijunzi Tang, Lizhong Tang and Fuzi Lizhong Tang could effectively promote the cell proliferation, and inhibit the cellular apoptosis of nonalcoholic fatty liver cells(P<0.01). And Fuzi Lizhong Tang showed the best effect. Western blot results showed that Sijunzi Tang, Lizhong Tang and Fuzi Lizhong Tang could down-regulate the expressions of cleaved caspase-3, cleaved caspase-8, cleaved caspase-9 and Bax proteins, and up-regulate the expressions of PCNA and Bcl-2 proteins of nonalcoholic fatty liver cells. And Fuzi Lizhong Tang showed the best effect. In conclusion, all of Sijunzi Tang, Lizhong Tang and Fuzi Lizhong Tang could effectively promote the cell proliferation, and inhibit the cellular apoptosis of nonalcoholic fatty liver cells. And Fuzi Lizhong Tang showed the best effect. The pharmacodynamic mechanism may be related to the expressions of key factors in pathways related with proliferation and apoptosis mediated by the three decoctions.

Structure features of polysaccharides from Chrysanthemum morifolium and their activities against tumor cells and NF-κB / 中草药

Objective: To clarify the structure features of polysaccharides from Chrysanthemum morifolium (PCM) and to study their activities against tumor cells and NF-κB. Methods: Six homogeneous neutral polysaccharides were obtained from three kinds of C. morifolium (Hangju, Huaiju, and Boju) flowers by successive hot water extraction, followed by ethanol precipitation, ion-exchange chromatography, and gel permeation chromatography. Their primary structures were characterized by HPGPC, IR, GC, and GC-MS analyses. Their bioactivities were examined by MTT assay using PANC-1 and LO2 cells. In addition, NF-κB signaling activation in PANC-1 and LO2 cells treated by polysaccharides were also measured. Results: The weight-average molecular mass of the six PCM, CMTA0S1, CMTA0S3, CMJA0S1, CMJA0S2, CMBA0S1, and CMBA0S3 was 7.523 × 104, 7.80 × 103, 7.80 × 104, 1.04 × 104, 5.79 × 104, and 1.35 × 104, respectively. CMTA0S1, CMJA0S1, and CMBA0S1 mainly contained galactose (Gal), arabinose (Ara) and glucose (Glc) residues in molar ratio of 1.23:1.00:0.20, 2.18:1.00:0.53, and 3.30:1.00:0.75, while CMTA0S3, CMJA0S2, and CMBA0S3 mainly contained Gal, Ara, Glc, and mannose (Man) residues in molar ratio of 0.73:1.00:0.40:0.21, 1.39:1.00:0.84: 0.55, and 1.19:1.00:0.48:0.19. Methylation analysis indicated that six PCM primarily consisted of T-arabinofuranosyl, 1, 5-arabinofuranosyl, 1, 4-galactopyranosyl, 1, 3, 6-galactopyranosyl, and 1, 4-glucopyranosyl residues. The biological activity study suggested that all the PCM could inhibit the growth of PANC-1 cells. Among them the inhibitory rates of CMTA0S3 and CMJA0S2 were at most to 70% with concentration-effect relationship. The NF-κB inhibition test indicated that only the crude polysaccharide CMBA had strong immunosuppressive activity, and homogeneous polysaccharides CMTA0S1 and CMJA0S1 showed potential immunostimulation. Conclusion: The six homogeneous polysaccharides share similar structures and inhibition on PANC-1 cells growth. Meanwhile they also may regulate the NF-κB activation.