ABSTRACT
OBJECTIVE： To establish a quantitative analysis of multi-components by singer marker （QAMS） for determining the contents of diosgenin， 5-hydroxymethylfurfural， vanillic acid， rutin， quercetin， kaempferol in Polygonati Rhizoma and its decoction piece. METHODS： HPLC external standard method was used to determine the contents of 6 components in Polygonati Rhizoma and its decoction piece simultaneously [the separation was carried out on Diamonsil-C18 column with mobile phase consisted of acetonitrile-water （gradient elution）； the detection wavelengths of 5-hydroxymethylfurfural， vanillic acid， rutin， quercetin and kaempferol were set at 254 nm（0-60 min）； the detection wavelength of diosgenin was set at 202 nm （60-75 min） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃]. Using vanillic acid as internal standard， relative correction factors （RCFs） of aother 5 components were calculated and to investigate durability. Relative retention method was used to accurately locate the chromatographic peaks of the components to be determined， and then the contents of the aother 5 components in Polygonati Rhizoma were calculated according to RCFs， and the results were compared with those determined by external standard method. The method was validated by Polygonati Rhizoma decoction piece. The contents of 6 components were determined by QAMS method and external standard method respectively， and then the differences of content determination were compared between 2 methods. RESULTS： The methodology investigation results of HPLC method were in line with related requirements. Within the linear range， the RCFs of diosgenin， 5-hydroxymethylfurfural， rutin， quercetin， kaempferol were 0.195， 0.025， 0.263， 0.345 and 0.075， respectively. Under different experiment conditions， RCFs showed good reproducibility； there was no statistical significance of 6 components in Polygonati Rhizoma and its decoction piece determined by external standard method and QAMS method （P＞0.05）. CONCLUSIONS： Established QAMS method is suitable for simultaneous determination of 6 components in Polygonati Rhizoma and its decoction piece.
ABSTRACT
Objective The objective of this study was to investigate the inhibitory effect of MDM2 inhibitor Nultin-3 com-bined with cisplatin on human oral squamous cell carcinoma(OSCC)Tca8113 cells and its mechanism. Methods Human OSCC Tca8113 cells were treated with Nultin-3,cisplatin,Nultin-3 combined with cisplatin,or vehicle control groups. The proliferation of Tca8113 cells was determined by thiazolyl blue(MTT)assay. The expression of MDM2,P53,Caspase-9 and Caspase-3 protein was determined by Western blot. Results The proliferative rate of OSCC ca8133 cells treated with Nultin-3 combined with cisplatin was significantly lower than that of other groups(P<0. 05). The relative expression of Caspase-9,Caspase-3 and P53 protein in the Nultin-3 combined with cisplatin was significantly higher than those of the Nultin-3 and cisplatin alone groups(P<0. 05). In addi-tion,the relative expression of MDM2 protein in the Nultin-3 combined with cisplatin group was significantly lower than that of the cisplatin and Nultin-3 alone groups(P<0. 05). Conclusion Nultin-3 combined with cisplatin has synergistic effect on oral squa-mous cell carcinoma Tca8113 cells. Nultin-3 regulates the MDM2-p53 signaling pathway and up-regulates the expression of pro-apoptotic proteins Caspase-9 and Casapase-3 to enhance the inhibition of cisplatin on oral squamous cell carcinoma,providing a solid theoretical basis for clinical research and treatment.