ABSTRACT
The aim of this paper was to discuss the effect of swertiamarin, gentiopicrin and sweroside on rheumatoid arthritis fibroblast-like synoviocytes(RA-FLSs) and B-cell lymphoma-2(Bcl-2) and their mechanisms. ZINC database and RCSB PDB database were retrieved for 3 D chemical structures of swertiamarin, gentiopicrin and sweroside and 3 D target protein structures. AutoDock Mgltools 1.5.6, AutoDockVina 1.1.2 and pyMOL 2.2.0 were applied for molecular docking to analyze the relationship between Bcl-2(1 GJH) target protein and important ingredients. The cell apoptosis of RA-FLSs was tested by Annexin V-FITC. The Bcl-2 protein expression of RA-FLSs treated with different ingredients was tested by Western blot. The Bcl-2 mRNA expression of RA-FLSs treated with different ingredients was tested by RT-PCR. Swertiamarin, gentiopicrin and sweroside were docked well with Bcl-2(1 GJH). The binding energy of swertiamarin was-6.9 kcal·mol~(-1), the binding energy of gentiopicrin was-6.7 kcal·mol~(-1) and the binding energy of sweroside was-6.4 kcal·mol~(-1). Compared with the blank group, the Bcl-2 protein expression of each group were reduced, while that of the gentiopicrin group was the highest(P<0.01). Compared with the blank group, the Bcl-2 mRNA expression of each groups were reduced. Gentiopicrin can reduce the Bcl-2 protein expression and the Bcl-2 mRNA expression, so as to promote the RA-FLSs apoptosis.
Subject(s)
Humans , Apoptosis , Arthritis, Rheumatoid/genetics , Cell Proliferation , Cells, Cultured , Fibroblasts , Iridoid Glucosides , Molecular Docking Simulation , Proto-Oncogene Proteins c-bcl-2/genetics , Pyrones , SynoviocytesABSTRACT
OBJECTIVE： To establish the method for the content determination of gentiopicrin and loganic acid in Gentiana scabra， and to investigate the correlation of their contents with appearance traits and quality gradation criterion. METHODS： HPLC method was adopted. The determination was performed on Ascentis Express C18 column with mobile phase consisted of 0.1％ phosphoric acid-acetonitrile （gradient elution） at the flow rate of 0.4 mL/min. The column temperature was 30 ℃， and detection wavelength was set at 240 nm. The sample size was 1 μL. Taking the length， number and diameter of fibrous roots as indexes， the appearance and morphological characteristics of G. scabra were studied. The relationship of gentiopicrin and loganic acid content with the appearance property of medicinal material was analyzed by SPSS 21.0 software. k-mean clustering analysis was carried out by using SPSS 21.0 software， and gradation standard for G. scabra was established preliminarily. RESULTS： The linear range of gentiopicrin and loganic acid were 0.5-3.0 μg/mL （r＝0.999 9） and 0.05-0.50 μg/mL （r＝0.999 9）. The limit of quantification of gentiopicrin and loganic acid were 0.295， 0.289 μg/mL； the detection limit were 0.082， 0.081 μg/mL； RSDs of precision， stability， repeatability tests were all lower than 2％； the recovery rates were 97.56％- 102.23％ （RSD＝1.56％， n＝6） and 97.58％-102.67％ （RSD＝1.86％， n＝6）. Correlation results showed that there was a significant positive correlation of the length of G. scabra， the number of roots， root diameter， with the contents of gentiopicrin and loganic acid. The order of affecting content was the number of roots ＞length ＞root diameter. k-means clustering analysis showed that 54 batches of G. scabra was divided into two categories； S4-S6，S13，S17-S23，S25，S28，S31-S34 were clustered into a category； S1-S3， S7-S12， S14-S16， S24， S26，S27，S29， S30，S35-S54 were clustered into the other category. The results of gradation showed that 54 batches of G. scabra could be divided into two grades， and the results were consistent with the cluster analysis. CONCLUSIONS： Established method is simple and stable， and can be used for simultaneous determination of gentiopicrin and loganic acid in G. scabra. The more fibrous roots， the longer the length， the thicker the root， the higher the content of gentiopicroside and loganic acid， the better the quality of G. scabra.
ABSTRACT
Objective To establish a method for qualitative and quantitative analysis of Zhendian Kaiqiao Granules. Methods Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba in the preparation were identified by TLC. Gentiopicrin and paeoniflorin were determined by HPLC. The analysis was performed on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm), with the mobile phase of methyl alcohol-water (12:88). The flow rate was 1.0 mL/min and the column temperature was maintained at room temperature; the detection wavelengths were set at 273 nm (gentiopicrin) and 230 nm (paeoniflorin). Results Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba could be detected by TLC. Gentiopicrin showed a good linear relationship at a range of 2.396–11.980 μg, r=0.999 6. The average recovery was 99.72%, and RSD was 2.70%. Paeoniflorin showed a good linear relationship at a range of 2.728–13.640 μg, r=0.999 0. The average recovery was 98.74%, and RSD was 2.42%. Conclusion The established method is simple, reliable and reproductive.The method can be used for control quality of Zhendian Kaiqiao Granules.
ABSTRACT
Objective To establish the quality standards of Longdan Xiegan Capsule and Longdan Xiegan Soft Capsule. Methods Gentianae Radix et Rhizoma, Bupleuri Radix, Scutellariae Radix, Gardeniae Fructus, Alismatis Rhizoma, Angelicae Sinensis Radix, Rehmanniae Radix and Glycyrrhizae Radix et Rhizoma were identified by TLC. Gentiopicrin in Gentianae Radix et Rhizoma, geniposide in Gardeniae Fructus, baicalin in Scutellariae Radix were determined by HPLC. Results The TLC spots developed were clear. Gentiopicrin showed good linear relationship in the range of 0.066 1-0.595 1 mg (r=1.000 0), the average recovery of Capsule was 99.34%(RSD=1.50%), and the average recovery of Soft Capsule was 96.62%(RSD=1.50%). Geniposide showed good linear relationship in the range of 0.076 1-1.369 8 mg (r=0.999 9), the average recovery of Capsule was 101.3% (RSD=1.70%), and the average recovery of Soft Capsule was 100.59%(RSD=0.79%). Baicalin showed good linear relationship in the range of 0.214 4-1.608 mg (r=1.000 0), the average recovery of Capsule was 101.12% (RSD=1.30%), and the average recovery of Soft Capsule was 98.07% (RSD=2.40%). Conclusion The method is simple and reproducible. It can be used to control the quality of Longdan Xiegan Capsule and Longdan Xiegan Soft Capsule.
ABSTRACT
OBJCTIVE:To determine the content of Gentiopicrin in Qianggan capsules by HPLC.METHODS:The sample was separated on Diamonsil TM C18 (200 mm?4.6 mm,5 ?m) with mobile phase consisted of methanol-water (23∶77) at a flow rate of 1.0 mL?min-1.The detection wavelength was set at 254 nm.RESULTS: The linear range of Gentiopicrin was 0.018 59～0.232 40 ?g(r=0.999 6) with an average recovery of 96.59%(RSD=1.35%,n=6).CONCLUSION: The meth-od developed is accurate,stable and reproducible,and it provides basis for the improvement of the quality standard of Qianggan capsules.
ABSTRACT
OBJECTIVE: To establish the method for the determination of emodin, resveratrol, polydatin and gentiopicrin in the Compound Polygonum cuspidatum semisolid matrix capsule in order to provide basis of its quality control. METHODS: TLC scanning method was used to determine emodin and resveratrol at same plate and the developer was petroleum ether-butyl acetate-methanol-glacial acetic acid(4∶1∶0.7∶0.03) with determination wavelength of 285 nm and 293 nm respectively. Polydatin and gentiopicrin were determined at same plate and the developer was chloroform-methanol(7 ∶ 2)with determination wavelength of 313 nm and 270 nm respectively. RESULTS: All target components were separated and the standard curves of emodin, resveratrol, polydatin and gentiopicrin showed good linearity. Average recoveries of them were 100.1%(RSD=0.86%),99.6%(RSD=2.86%), 101.4%(RSD=0.99%),98.5%(RSD=2.89%),respectively. CONCLUSION: Established method is rapid, simple for the quality control of Compound P. cuspidatum semisolid matrix capsule.
ABSTRACT
OBJECTIVE:To optimize the extraction craft of gentiopicrin from gentian.METHODS:The extraction craft was optimized by orthogonal test with gentiopicrin as an index,and the content of gentiopicroside was determined by TLC scanning method.RESTLUTS:The quantity of menstruum and the extraction times of gentiopicrin had significant influence in the gentiopicrin extraction(P
ABSTRACT
Objective To establish a proper extraction method for Yangxue Ruanjian Prescription (YRP) by comparing the chemical component contents and analgesic and anti - inflammatory effect of extracts extracted from YRP by different methods. Methods Water - decocting, alcohol - refluxing and alcohol - percolating methods were adopted and component indexes of paeoniflorin and gentiopicrin were determined by HPLC. The anti - inflammatory effect of the extracts were observed on mice with xylol - induced auricular swelling and analgesic effect by hot - plate method. Results The content of gentiopicrin is similar in the extracts extracted by three methods and the content of paeoniflorin is higher in alcohol - percolating extract than in alcohol - refluxing extract, lowest in water - extraction. The analgesic effect of water - decocting extract is better than that of alcohol - refluxing extract, and that of alcohol - percolating extract is weakest; the anti - inflammatory effect of the three extracts is almost the same. Conclusion Water - decocting extraction is suitable to YRP, and this is determined by the characteristics of Chinese compound prescription.
ABSTRACT
AIM: To establish the method for determination of gentiopicrin, palmatine hydrochloride and berberine hydrochloride in Ertong Ⅱ Oral Liquid (Cortex Phellodendri, Radix Gentianae, Rhizoma Anemarrhenae, etc.) by RP HPLC. METHODS: The column was a Diamonsil TM C 18 column (250mm?4.6mm, 5?m); the mobile phase was CH 3CN H 3PO 4 C 6H 15 N H 2O; The ultraviolet detection was set at 270nm; the flow rate was 1mL?min -1 . RESULTS: The linear ranges of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 72.8～728?g?mL -1 ( r=0.9998 ), 7.55～75.5?g?mL -1 ( r=0.9998 ), 12.45～124.5?g?mL -1 ( r = 0.9999 ), respectively. The average recoveries ( n =6) of gentiopicrin, palmatine hydrochloride and berberine hydrochloride were 98.17%, 98.78%, 98.60%; RSD were 1.60%, 1.35%, 1.25%, respectively. CONCLUSION: The method is accurate, reproducible and highly selective, thus suitable for quality control of Er Tong Ⅱ Oral Liqual.
ABSTRACT
Objective: Enriching secoiridoid glycoside in Gentiana manshurica kitag. Methods: Gentiana manshurica kitag. was extracted with 70% EtOH by cold macerating and purified with AB 8 macroreticular resin. Results: The content of gentiopicrin in the extract is 28.69%. Conclusion: AB 8 macroreticular resin fits in purification of water soluble secoiridoid glycoside in Gentiana manshurica kitag.
ABSTRACT
Objective: To establish the quality standards for Xiegananshen Capsules. Methods: Poria in this prescription was identified by microscopic examination; Radix Gentianae, Fructus Gardeniae, Radix Angelicae Sinensis and Radix Scutellariae were identified by TLC. Gentiopicrin was determined by HPLC. Results: Poria could be detected by microscopic examination; Radix Gentianae, Fructus Gardeniae, Radix Angelicae Sinensis and Radix Scutellariae could be detected by TLC. Gentiopicrin showed a good linear relationship at a range of 2.6 ～13?g, r=0.9995. The average recovery was 97.8%, and RSD was 1.0%. Conclusion: The established methods are simple, feasible and reproducible. This study provided a method for the quality control of Xiegananshen Capsules.