Synergistic anti-liver cancer effects of curcumin and total ginsenosides

BACKGROUND Liver cancer is the sixth most frequently occurring cancer in the world and the fourth most common cause of cancer mortality.The pathogenesis of liver cancer is closely associated with inflammation and immune response in the tumor microenvironment.New therapeutic agents for liver cancer,which can control inflammation and restore cellular immunity,are required.Curcumin(Cur)is a natural anti-inflammatory drug,and total ginsenosides(TG)are a commonly used immunoregulatory drug.Of note,both Cur and TG have been shown to exert anti-liver cancer effects.AIM To determine the synergistic immunomodulatory and anti-inflammatory effects of Cur combined with TG in a mouse model of subcutaneous liver cancer.METHODS A subcutaneous liver cancer model was established in BALB/c mice by a subcutaneous injection of hepatoma cell line.Animals were treated with Cur(200 mg/kg per day),TG(104 mg/kg per day or 520 mg/kg per day),the combination of Cur(200 mg/kg per day)and TG(104 mg/kg per day or 520 mg/kg per day),or 5-fluorouracil combined with cisplatin as a positive control for 21 d.Tumor volume was measured and the protein expression of programmed cell death 1 and programmed cell death 1 ligand 1(PD-L1),inflammatory indicators Toll like receptor 4(TLR4)and nuclear factor-κB(NF-κB),and vascular growth-related factors nitric oxide synthases(iNOS)and matrix metalloproteinase 9 were analyzed by Western blot analysis.CD4+CD25+Foxp3+regulatory T cells(Tregs)were counted by flow cytometry.RESULTS The combination therapy of Cur and TG significantly inhibited the growth of liver cancer,as compared to vehicle-treated animals,and TG showed dose dependence.Cur combined with TG-520 markedly decreased the protein expression of PD-L1(P<0.0001),while CD4+CD25+Foxp3+Tregs regulated by the PD-L1 signaling pathway exhibited a positive correlation with PD-L1.Cur combined with TG-520 also inhibited the cascade action mediated by NF-κB(P<0.0001),thus inhibiting the TLR4/NF-κB signalling pathway(P=0.0088,P<0.0001),which is associated with inflammation and acts on PD-L1.It also inhibited the NF-κB-MMP9 signalling pathway(P<0.0001),which is associated with tumor angiogenesis.CONCLUSION Cur combined with TG regulates immune escape through the PD-L1 pathway and inhibits liver cancer growth through NF-κB-mediated inflammation and angiogenesis.

Dynamic Microwave-Assisted Extraction of Total Ginsenosides from Ginseng Fibrous Roots

A dynamic microwave-assisted extraction（DMAE） method is established for the extraction of total ginsenosides from ginseng fibrous roots. The extraction process has been simulated and its main affection factors（liquid/solid ratio K of solvent to ginseng powders（V/m）, irradiation time, irradiation temperature, extracting solution concentration, flow rate of solvent and microwave power） have been optimized by response surface methodology（RSM）. The optimum conditions of extraction are the liquid/solid ratio of 270 m L/g, the extraction temperature of 75 ℃, the extraction time of 35 min, ethanol concentration of 70%（V/V）, the solvent flow rate of 1.3 m L/min, and the microwave power of 500 W. The yield of ginsenosides obtained by the proposed DAME method is（15.0±0.7） %, which is well agreement with the yield predicted by the model. Compared with static microwave-assisted extraction, DMAE has a higher extraction yield and can avoid the degradation of ginsenoside.

Profiling and identification of chemical components of Shenshao Tablet and its absorbed components in rats by comprehensive HPLC/DAD/ESI-MS^n analysis

Shenshao Tablet(SST),prepared from Paeoniae Radix Alba(PRA)and total ginsenoside of Ginseng Stems and Leaves(GSL),is a traditional Chinese medicine(TCM)preparation prescribed to treat coronary heart disease.However,its chemical composition and the components that can migrate into blood potentially exerting the therapeutic effects have rarely been elucidated.We developed an HPLC/DAD/ESI-MS^n approach aiming to comprehensively profile and identify both the chemical components of SST and its absorbed ingredients(and metabolites)in rat plasma and urine.Chromatographic separation was performed on an Agilent Eclipse XDB C_(18) column using acetonitrile/0.1% formic acid as the mobile phase.MS detection was conducted in both negative and positive ESI modes to yield more structure information.Comparison with reference compounds(t_R,MS^n),interpretation of the fragmentation pathways,and searching of in-house database,were utilized for more reliable structure elucidation.A total of 82 components,including 21monoterpene glycosides,four galloyl glucoses,two phenols from PRA,and 55 ginsenosides from GSL,were identified or tentatively characterized from the 70% ethanolic extract of SST.Amongst them,seven and 24 prototype compounds could be detectable in the plasma and urine samples,respectively,after oral administration of an SST extract(4 g×kg^(–1))in rats.No metabolites were observed in the rat samples.The findings of this work first unveiled the chemical complexity of SST and its absorbed components,which would be beneficial to understanding the therapeutic basis and quality control of SST.