ABSTRACT
Objective: To develop the quality standard for Shenmai Dihuang Pills (SDP). Methods: The TLC methods applied to identify MoutanCortex, CorniFructus, Alismatis Rhizoma, and GlehniaeRadix in the formulation were carried out according to the methods recorded in general rules of Chinese Pharmacopeia (2015 edition, volume 4). The contents of psoralen, iso psoralen, and paeonol were analyzed by high performance liquid chromatography on a Diamonsil C18 column (250 mm ×4.6 mm, 5μm) with the mobile phase of acetonitrile (A)-0.1% formic acid (B) by gradient elution at a flow rate of 1.0 mL/min. The detection wavelength was set at 254 nm and the column temperature was controlled at 30℃. Results: There were good specificities of TLC for MoutanCortex, CorniFructu, Alismatis Rhizoma, and Glehniae Radix. The methodology validation for the assay of psoralen, iso psoralen, and paeonol presented that they were in good linear correlation in the ranges of 6.531-58.78, 6.115-55.04 and 20.40-183.6 μg/mL, with the regression equations of Y =1.523×104X-1.209×104 (r = 0.999 9), Y =1.809×104 X-1.523×104 (r = 0.999 9), and Y =2.760 ×104 X-1.683 ×104 (r = 0.999 9), respectively. The average recoveries were 100.4% (RSD 1.1%), 101.5% (1.3%), and 101.5% (0.79%).The content ranges of psoralen, isopsoralen, and paeonol from five different batches of SDP were 322.1-331.0, 298.0-350.1, and 929.5-982.7 μg/g, respectively. Conclusion: The established quality of TLC and HPLC methods are specific, reliable, accurate, and suitable, which can be successfully applied to the quality control for the preparation.
ABSTRACT
To study the inhibitory effect of Glehniae Radix petroleum ether part on TGF-β1-induced epithelial mesenchymal transition in non-small cell lung cancer A549 and its possible mechanism. With type Ⅱ epithelial cells of lung cancer A549 as the research object, the experiment was performed in 5 μg•L⁻¹ TGF-β1-induced epithelial mesenchymal transition model,and blank control group, model group and Glehniae Radix petroleum ether group were set up. MTT assay was carried out to detect the effect of petroleum ether extract of Glehniae Radix on the survival of A549 cells. A549 cells induced by TGF-β1(5 μg•L⁻¹) was intervened by different polar parts of Glehniae Radix, Real-time quantitative polymerase chain reaction(RT-qPCR) was used to analyze mRNA expressions of the epithelial mesenchymal transition markers, such as ColⅠ,E-cadherin,Vimentin and α-SMA. Enzyme linked immunosorbent assay(ELISA) was used to detect hydroxyproline(HYP) level. The migration and invasion abilities of cells were detected through wound scratch assay. According to the experimental results, the petroleum ether extract of Glehniae Radix could inhibit the growth of A549 cells in a concentration-dependent manner. Compared with model group, Glehniae Radix petroleum ether part group could effectively inhibit mRNA expressions of ColⅠ,Vimentin and α-SMA, but improve expression of E-cadherin.Glehniae Radix petroleum ether part could reduce the content of hydroxyproline in cells and inhibit the migration of A549 cells.Therefore, the petroleum ether extract of Glehniae Radix can effectively inhibit the occurrence of epithelial mesenchymal transition induced by TGF-β1 induced alveolar epithelial cells, and Glehniae Radix petroleum ether part may be a potential drug for idiopathic pulmonary fibrosis. The mechanism may be achieved through the regulation of ColⅠ, Vimentin, α-SMA and E-cadherin.
ABSTRACT
Water-soluble polysaccharides from traditional Chinese medicine have properties of complex structure and high molecular, resulting in hardly complete their structural characterization.However, a "bottom-up" approach could solve this problem.Glehniae Radix extract was extracted with hot water and then precipitated by 40% ethanol to obtain Glehniae Radix polysaccharides (RGP). Subsequently, a partial acid hydrolysis method was carried out and the effects of acid concentration, time and temperature on hydrolysis were investigated. Under the optimum hydrolysis condition (1.5 mol•L⁻¹ trifluoroacetic acid, 4 h, and 80 ℃), RGP were hydrolyzed to characteristic oligosaccharide fragments. Futher, a hydrophilic liquid chromatography- mass spectrometry method was used for the separation and structural characterization of the polysaccharide hydrolysates. According to MS and MS/MS analysis of several standard disaccharides, a method for determining the type of polysaccharide glycosidic linkage by mass spectrometry was established. The results showed that the polysaccharide hydrolysates were linear glucan containing 1, 4-glycosidic bonds. And gluco-oligosaccharides with the degrees of polymerization (DP) of 4-11 were obtained after partial acid hydrolysis.