ABSTRACT
OBJECTIVE：To establish pre -column derivatization-HPLC fingerprint of Polyporus polysaccharide ，and to determine the contents of main monosaccharide components ，so as to provide reference for quality evaluation of Polyporus umbellatus. METHODS ：Polyporus polysaccharide was extracted with boiling water and precipitated by ethanol and deproteinized by Sevage from 11 batches of P. umbellatus from different producing areas. The samples were firstly hydrolyzed with trifluoro-acetic acid （TFA）and then derivatized by 1-phenyl-3-methyl-5-pyrazolone（PMP）. HPLC analysis was then conducted. The determination was carried out on HypersiL BDS C 18 column with mobile phase composed of 0.1 mol/L phosphate buffer （pH 6.84）-acetonitrile（84∶16，V/V）by gradient elution at the flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm， and column temperature was 30 ℃. The sample size was 20 µL. The similarity of 11 batches of Polyporus polysaccharide was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System （2012A edition ），and the contents of main monosassharide components were detected. The peak was identified by comparing with the reference substance ，and cluster analysis was performed by using SPSS 23.0 software. RESULTS ：In HPLC fingerprints of the 11 batches of samples ，3 common peaks were identified ，namely mannose ，glucose and galactose. The similarity of all samples was above 0.94. Cluster analysis classified 11 batches of samples into three categories. S 1-S6，and S 8 were grouped into category 1；S7，S10 and S 11 were grouped into category 2；S9 was individually grouped into one category. Results of content determination showed that the contents of mannose ranged from 1.571 to 8.771 mg/g；those of glucose ranged from 26.072 to 132.194 mg/g，and those of galactose ranged from 3.420 to 36.593 mg/g. CONCLUSIONS ：Established pre-column derivatization HPLC fingerprints can provide reference for quality evaluation of P. umbellatus . The monosaccharide composition of different batches of Polyporus polysaccharide is the same ；there is no significant correlation between fingerprint characteristic peak and the origin of herbs ；there is significant difference in the content of monosaccharide of P. umbellatus .
ABSTRACT
Objective: To study the effects of polyporus polysaccharide (PPS) on the proliferation of human lung cancer A549 cells and the corresponding molecular mechanism. Methods: The A549 cells in control group were normally treated and the cells in experimental groups were incubated with different doses of PPS. The MTT method and flow cytometry were used to analyze the influence of drugs on cell proliferation. The qRT-PCR was used to detect mRNA levels of A549 cells and Cyclin D1 mRNA stability. The levels of human antigen R (HuR) protein in plasma and nuclei and cyclin D1 protein were detected by Western blotting. Results: Compared with the control, the cell growth was obviously inhibited (P<0.05), the Cyclin D1 mRNA stability and Cyclin D1 mRNA were decreased in the mid-and high-dose PPS groups (P<0.05), the total protein of HuR was slightly changed, but the cytoplasm protein of HuR was down-regulated (P<0.05) and the nucleus protein of HuR was up-regulated (P<0.05). Conclusion: PPS can suppress the proliferation of A549 cells, which is possibly affected by down-regulating the cytoplasm protein of HuR, lowering the stability of Cyclin D1 mRNA, and thus decreasing the expression of Cyclin D1 protein.
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1.0 mg?L -1 ),the intracellular calcium became increased,and kept on a higher level.Conclusion PUPS participates in the process of calcium message transduction.