Effects of Monoammonium Glycyrrhizinate on Stem Cell-like Characteristics, Oxidative Stress and Mitochondrial Function of Acute Promyelocytic Leukemia NB4 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of monoammonium glycyrrhizinate on the stem cell-like characteristics, oxidative stress and mitochondrial function of acute promyelocytic leukemia cells NB4.@*METHODS@#CCK-8 method was used to detect the viability of acute promyelocytic leukemia cells NB4, and the appropriate dose was screened; Cloning method was used to detect the proliferation rate of NB4 cell; Western blot was used to detect the expression of cell cycle-related protein; flow cytometry was used to detect cell apoptosis and sort NB4 stem cells positive (CD133+); Stem cell markers (Oct4, ABCG2, Dclk1) were detected by RT-PCR; ROS was detected by fluorescence; The kit was used to detect the level of oxidative stress markers (MDA); The flow cytometry was used to detect the change of mitochondrial membrane potential; Western blot was used to detect the expression of mitochondrial damage index-related proteins (Bax/BCL-2).@*RESULTS@#Compared with the control group, if the concentration of MAG was less than 5 μmol/L, the cell NB4 viability showed no significant difference; if the concentration was higher than 5 μmol/L, the inhibitory effect on the growth of cell NB4 increased and showed significant difference (P<0.05), according to the results of CCK-8 experiment, four groups were set based on the concentration of MAG 0 μmol/L, MAG 5 μmol/L, MAG 10 μmol/L, and MAG 20 μmol/L; compared with the control group (MAG 0 μmol/L), the cells in MAG 5 μmol/L group showed no significant difference, while the proliferation rate, cyclin expression, mitochondrial membrane potential, stem cell CD133+ ratio, and marker mRNA level ( Oct4, ABCG2, Dclk1) of NB4 cell were significantly reduced (P<0.05); the apoptosis rate, reactive oxygen species, MDA content and Bax/BCL-2 expression of NB4 cell significantly increased (P<0.05).@*CONCLUSION@#Monoammonium glycyrrhizinate has a significant inhibitory effect on acute promyelocytic leukemia cells NB4, which may be related to the regulation of stem cell-like characteristics, oxidative stress and mitochondrial function.

Effect of different decocting methods on active ingredients in Banxia Xiexin Decoction / 中草药

Objective To investigate the effects of the three methods of decocting with deslag, decocting without deslag, and double decocting on the content of nine ingredients baicalin, baicalein, ginsenoside Re, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, liquiritin, 6-gingerol, berberine hydrochloride, palmatine hydrochloride, and total flavonoids in Banxia Xiexin Decoction (BXD). Methods Nine index components were determined by HPLC. The HPLC analysis was performed on Welch Ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) with mobile phase of acetonitrile-0.1% phosphate aqueous solution for gradient elution; And carried out at column temperature of 28 ℃, volume flow of 0.9 mL/min, and detection wavelength of 203, 252, 280, and 355 nm. The total flavonoids were determined by colorimetry. Results Nine kinds of ingredients and total flavonoids could be detected in three different decoctions. In the method of decocting with deslag, baicalin, baicalein, ginsenoside Rb1, monoammonium glycyrrhizinate hydrate, and liquiritin increased by 10.01%, 12.88%, 29.09%, 16.75%, and 15.02%, respectively, compared with decocting without deslag; It decreased by 5.54%, 4.15%, 14.49%, 7.85%, and 9.18%, respectively compared with double decocting; Ginsenoside Re, 6-gingerol, berberine hydrochloride, and palmatine hydrochloride increased by 37.90%, 3.78%, 5.33%, and 5.99% compared with decocting without deslag, respectively; compared to the double decocting methods, it increased by 1.07%, 11.57%, 3.41%, and 1.93%. The total flavonoids increased 22.61% higher than decocting without deslag and 6.54% higher than double decocting. Conclusion: The results can effectively reflect the quality difference of different decocting methods. Among the three methods of decoction, the method of decocting without deslag has significantly improved the dissolution of the active ingredients of each component in the decoction, and improve the clinical efficacy of BXD to a certain extent. It provides a good experimental basis for the decocting without deslag method used in Zhang Zhongjing’s Treatise on Febrile Diseases.

Migration study on ceramic membrane separation process of active ingredient population in compound Chinese materia medica / 中草药

Objective: Taking thewater extraction of Compound Banlangen Liyan Granules (IsatidisRadix, ScrophulariaeRadix, PlatycodiRadix, GlycyrrhizaeRadix, etc.)as the research object to explore the dynamic migration regular of the active ingredient population of compoundChinesemateriamedica in different apertures in ceramic membrane separation process and compare its membrane filtration flux and solid inclusion removal rate. Methods: The water extraction of Compound Banlangen Liyan Granules was separated with 200 nm and 50 nm aperture of ceramic membrane, respectively. Continuously sampling for many times was done in the ceramic membrane separation process. The optimized HPLC methods were as follows: chromatographic column was Agilent Zorbox Eclipse XDB-C18, mobile phase was acetonitrile-0.05% phosphoric acid in gradient elution at flow rate of 1 mL/min, detection wavelength was wavelength switching, injection volum was 10 μL, the column temperature was 30 ℃. The contents of sixactive ingredients[adenosine, (R,S)-goitrin, liquiritin, harpagosid, platycodinD, and monoammonium glycyrrhizinate]were determined in thecompound at the same time, and the dynamic migration rates were investigated. Results:The simultaneous determination of the sixactive ingredient in thecompound was done with HPLC method. For the water extraction in Compound Banlangen Liyan Granules, in the 200 nm ceramic membrane separation process, the dynamic migration rates of the sixactive ingredientsranged between 71%-104%, the average migration rate was 85%, the membrane filter flux attenuation was smaller, the stable flux was in 426-340 L/(m2∙h), solid inclusion removal rate was 21.0%. But in 50 nm ceramic membrane, the dynamic migration rates of the sixactiveingredients ranged between 83%-107%, the average migration rate was83%, the membrane filter flux attenuation was larger, the stable flux was in 258-228 L/(m2∙h), and solid inclusion removal rate was23.9%. Conclusion :The experiment preliminarily reflects the dynamic migration regular of the active ingredient population of compoundChinesemateriamedica in two different aperture ceramic membrane separation process. It lays a migration theoretical foundation of effective materials for popularization and application of modern ceramic membrane technology in Chinese materiamedica refining.

Quality standard for Yiganjian Tablets / 中成药

AIM: To establish the methods of identifing and determining Yiganjian Tables. METHODS: Herba Halenia,extract of Radix et Rhizoma Glycyrrhizale,Radix Astragali were identified by TLC.The content of 1-hydroxy-3,4,5-trimethoxyxanthon in Herba Halenia was determined by HPLC on Hypersil-ODS2 column with the mobile phase of methanol-0.2%H_3PO_4(55∶45).The detective wavelength was set at 243 nm and monoammonium glycyrrhizinate was determinad by HPLC on Hypersil-ODS2 column with the mobile phase of acetonitrile-2% aceacid(60∶40).The detective wavelength was set at 250 nm. RESULTS: The linear range of 1-hydroxy-3,4,5-trimethoxyxanthon was from 0.173 2 to 0.866 0 ?g.The average recovery was 100.56%,RSD was 2.7%(n=9).The linear range of monoammonium glycyrrhizinate was from 0.55 to 2.75 ?g.The average recovery was 98.29%,RSD was 1.7%(n=9). CONCLUSION: The method is simple,accurate and sensitive,so it can be used for the quality control of Yiganjian Tablets.