Improvement Effects of Alisol B 23-acetate on Glycolipid Metabolism Disorder in Obesity Model Mice / 中国药房

OBJECTIVE：To st udy the improvement effects and its mechan ism of alisol B 23-acetate on glycolipid metabolism disorder in obesity model mice. METHODS ：The mice was given high-fat diet for 10 weeks to induce obesity model. Model mice were randomly divided into model group ，orlistat group （positive control ，15.6 mg/kg）， alisol B 23-acetate low-dose ， medium-dose and high-dose groups （7.5，15，30 mg/kg），with 10 mice in each group. Another 10 mice fed with normal diet were set as normal group. The mice in normal group and model group were given water intragastrically ，and administration groups were given the corresponding drugs intragastrically ，with the volume of 20 mL/kg，once a day ，for consecutive 4 weeks. After last medication，body weight ，waist circumference ，body fat ，muscle and body fluid mass were measured ；the serum levels of blood lipids indicators （TC，TG，HDL-C，LDL-C）and blood glucose were determined. The levels of PPAR-γ，NF-κB and IL-6 in liver tissue as well as serum level of TNF-α were determined by ELISA. The pathomorphological changes of visceral fat and liver tissue in mice were observed by HE staining. RESULTS ：Compared with normal group ，body weight ，waist circumference ，body fat and body fluid mass were significantly increased in model group （P＜0.01）；serum levels of TC ，TG，HDL-C，blood glucose and TNF-α，the levels of PPAR-γ，NF-κB and IL-6 in liver tissue were increased significantly （P＜0.05 or P＜0.01）；the structure of adipocytes was ruptured ，the volume of adipocytes was increased ，accompanied by inflammatory cell infiltration ；a large number of liver cells were edema ，and cytoplasm was loose and light stained，accompanied by fatty degeneration. Compared with model group ，the body weight ，body fat and body fluid mass as well as serum le vels of TG and TNF-α in alisol B 23-acetate groups were significantly reduced （P＜0.01）；the levels of TC and blood glucose in serum ，IL-6 in liver tissue were significantly decreased in alisol B 23-acetate medium-dose and high-dose groups （P＜0.05 or P＜0.01），and the level of PPAR-γ in liver tissue was increased significantly（P＜0.05 or P＜0.01）； the waist circumference and NF-κB levels in liver tissue in alisol B 23-acetate high-dose group were decreased significantly （P＜ 0.01）；serum level of HDL-C in alisol B 23-acetate medium-dose group were decreased significantly （P＜0.01）；the adipocytes were closely arranged and small in size ；the hepatocytes were mild to moderate swelling ，a small amount of cytoplasm was loose ， light stained or vacuolated ，and a small number of hepatocytes were accompanied by steatosis and small focal infiltration of inflammatory cells. CONCLUSIONS ：Alisol B 23-acetate can improve the disorder of glucose and lipid metabolism in obesity model mice ，and its mechanism may be related to the regulation of PPAR-γ，NF-κB，IL-6 levels in liver tissue and TNF-α levels in serum.

Analysis on material basis and efficacy network of Huopo Xialing Decoction in treating damp pathogen stagnation of lung syndrome of early COVID-19 / 中草药

Objective: To screen the material basis of Huopu Xialing Decoction in the treatment of damp pathogen stagnation of lung syndrome of early COVID-19 and predict its mechanism. Methods: Literatures and clinical reports were reviewed to analyze the relationship between Huopu Xialing Decoction and damp pathogen stagnation of lung syndrome of early stage of COVID-19. TCMSP database was used to screen the potential active components in Huopo Xialing Decoction. Molecular docking of the active components with SARS-CoV-2 hydrolase and ACE2 was also carried out. According to the binding ability, the core components with both were screened. The interaction network of key components target proteins was constructed by using the software of Cytoscape to screen the main targets; The GO analysis of the main targets was carried out by using the STRING database, and the Pathway and KEGG enrichment analysis was carried out by using the plug-in of the software ClueGO of Cytoscape. Results: The Huopo Xialing Decoction was used to treat the early coronavirus pneumonia with damp pathogen and lung depression syndrome in the relationship analysis between prescriptions and syndrome, and the potential components of the 12 ingredients in Huopo Xialing Decoction were selected, with 67 core targets. Among them, paryriogenin I from Tetrapanax papyrifer, ergosterol peroxide from Poria cocos and Polyporus umbellatus, baicalin from Pinellia ternata had good binding activity with SARS-CoV-2 3CL hydrolase and ACE2. The results of enrichment analysis of GO, Pathway and KEGG showed that 12 potential components in Huopo Xialing Decoction were involved in regulating the biological processes such as stimulation response, signal transduction, cell death and the related signaling pathways including interleukins, EGFR in cancer, tyrosine kinases, programmed cell death and MAPK signaling pathways. Conclusion: For the early COVID-19 patients with the syndrome of damp pathogen stagnation of lung, Huopo Xialing Decoction was used to resolve dampness and detoxification, and ventilate lung and promote pathogenic penetration. Phenanthrone, baicalin, jujuboside_qt, cerevisterol, hederagenin, ergosterol peroxide, citrostadienol, ergosterol-7,22-diene-3-one, paryriogenin I, alisol B,23-acetate, alisol B, neohesperidin may be the main material basis, and play a role by blocking the protein synthesis of SARS-CoV-2 virus, preventing the virus from entering the host cells, regulating the IL signaling pathway, MAPK signaling pathway, PI3K-Akt signaling pathway, T cell receptor signaling pathway, C-type lectin receptor signaling pathway and inhibiting the expression of related inflammatory factors.

Preparation and quality evaluation of standard decoction of Alismatis Rhizoma / 中草药

Objective: To prepare standard decoction of Alismatis Rhizoma (AR) and establish its quality evaluation system, and provide reference for the development of dispensing granules of AR. Methods: A total of 18 batches of AR decoction pieces were collected to prepare standard decoction of AR according to the standard process. Quality evaluation system of standard decoction of AR was established with pH value, dry extract rate, fingerprint similarity and transfer rate of alisol B 23-acetate as indexes. Results: The mass fraction of alisol B 23-acetate in AR decoction pieces was 0.057%—0.267% with the average value of 0.156%, water content was 9.2%—12.8% with the average value of 10.44%; the pH value of standard decoction of AR was 4.11—5.60, dry extract rate was 10.25%—17.09%; transfer rate of alisol B 23-acetate from decoction pieces to standard decoction was 10.49%—17.49%. Conclusion: The established quality evaluation method is stable and feasible, which is suitable for the development and quality evaluation of standard decoction of AR, which can provide reference for the development of dispensing granules of AR and related classic formulas.

Research progress of Alismatis Rhizoma and prediction analysis on its Q-marker / 中草药

Alismatis Rhizoma was first published in the “Shennong’s Classic of Materia Medica”, listed as top grade, and most of the ancient herbals have been collected. Alismatis Rhizoma is mainly distributed in Fujian, Sichuan, and Jiangxi provinces. Alismatis Rhizoma has the effect of diffusing water and dampness, releasing heat, removing turbidity, and reducing lipid. Its modern pharmacological activity is extensive, and its clinical research is deepening gradually. This paper summarizes the research status of Alismatis Rhizoma from the aspects of herbal textual research, chemical composition, and pharmacological action. On this basis, the differences of different radicals of Alismatis Rhizoma are clarified. Based on the concept of quality marker (Q-marker), the Q-markers of Alismatis Rhizoma are predicted from the point of origin, combined with the research of pharmacodynamics, new medicinal uses and processing, in order to perform qualitative and quantitative analysis of the effective components of Alismatis Rhizoma and provide scientific basis for formulation of new standards.

Exploration of objective quality evaluation parameter of Alismatis Rhizoma decoction pieces / 中国中药杂志

To establish a quality constant evaluation system of Alismatis Rhizoma decoction pieces,in order to provide reference for regulating the market circulation of this decoction pieces. A total of 18 batches of Alismatis Rhizoma decoction pieces were collected from different pharmaceutical factories,and the morphological parameters of each sample were tested. The content of alisol B 23-acetate in Alismatis Rhizoma decoction pieces was determined by HPLC in the 2015 edition of Chinese Pharmacopoeia,and the parameters such as quality constant and relative quality constant were calculated. The quality constant range of 18 batches of Alismatis Rhizoma decoction pieces was 0. 390-2. 076. If 18 batches of Alismatis Rhizoma decoction pieces were divided into 3 grades,taking 80% of the maximum quality constant as first grade,50% to 80% as second grade,and the rest as third grade,then the quality constant of firstgrade samples was ≥1. 66,the quality constant of second-grade samples was ≥1. 04 and <1. 66,and the quality constant of third-grade samples was <1. 04. The established quality constant evaluation method is objective and feasible,which can be used to classify the grade of Alismatis Rhizoma decoction pieces and provide a reference method to control the quality of this decoction pieces.

Influence of alisol B 23-acetate on blood glucose in type 2 diabetic mice / 中国药理学通报

Aim To explore whether alisol B 23-acetate possesses the therapeutic potential for treatment of type 2 diabetes mellitus ( T2DM ). Methods T2DM mouse model was established by combined administration of streptozotocin and nicotinamide. After three weeks of oral administration of rosiglitazone or alisol B 23-acetate, the blood glucose of type 2 diabetic mice was measured. Oral glucose tolerance test(OGTT) was carried out the next day. Rosiglitazone was chosen as positive drug. 2-[N-(7-nitrobenz-2-oxa-l, 3-diazol-4-yl) amino]-2-deoxy-D-glucose (2-NBDG) uptake assay in adipocytes was adopted to test whether alisol B 23-acetate had effect on glucose uptake in cells. 3T3-Ll pre-adipocytes differentiation model was performed to evaluate whether alisol B 23-acetate promoted adipo-genesis. Results Mice exhibited significantly higher blood glucose concentration after intraperitoneal injection of streptozotocin and nicotinamide for three weeks, as examined by blood glucose concentration on day 21 and OGTT on day 22, compared with normal mice in blank control group. After orally administrating alisol B 23-acetate at dose of 5 mg • kg-1 , 10 mg • kg-1 ,20 mg • kg-1 daily for three weeks, respectively, or orally administered rosiglitazone at dose of 10 mg • kg-1 daily for three weeks, blood glucose greatly decreased in type 2 diabetic mice. Moreover, insulin resistance was also improved to a certain degree during OGTT. Furthermore, alisol B 23-acetate not only increased insulin-induced glucose uptake in adipocytes at the concentration of 30 mmol • L-1 glucose, but also accelerated 3T3-L1 pre-adipocytes differentiation process at concentration of 1 μmol • L-1 and 10 μmol • L-1 . Conclusions Alisol B 23-acetate reduces blood glucose of type 2 diabetic mice, promotes pre-adipocyte differentiation and increases glucose uptake in adipocytes; however, the mechanism of action needs further exploration.

Research on effective components compatibility of Alismatis Rhizoma resisting calcium oxalate calculus in vitro based on uniform design / 中草药

Objective: To investigate the inhibitory effect of six active triterenoids and their compatibility from Alisma orientale on the formation of urinary calcium oxalate calculus in vitro. Methods: A uniform design method was used to design the different compatibilities from six triterenoids. The inhibitory effects of the six triterenoids along with their different compatibility groups were evaluated in vitro using a standard seeded crystallization technique. Results: The six active triterenoids and their compatible preparations could significantly inhibit calcium oxalate crystal formation in vitro (P < 0.05), and the group of alisol A-alisol A 24-acetate-alisol B-alisol B 23-acetate-alisol F-alisol F 24-acetate (2.2∶3.8∶1∶3.5∶2.2∶1) was the most efficient with an inhibitory index of 188.29%. Conclusion: The triterenoids in A. orientale play a key role in the inhibition of calcium oxalate calculus formation and compatibility of alisol A, alisol A 24-acetate, alisol B, alisol B 23-acetate, alisol F, and alisol F 24-acetate can inhibit the calcium oxalate calculus well in vitro especially cooperated with each other, and the best compatibility proportion is 2.2∶3.8∶1∶3.5∶2.2∶1, respectively.

Optimization of flash-type extraction technology of alisol B 23-acetate from Alismatis Rhizoma by response surface methodology / 中国中药杂志

Response surface methodology was used to optimize and obtain the optimal flash-type extraction technology of alisol B 23-acetate from Alismatis Rhizoma. With the extraction rate of alisol B 23-acetate as an indicator, single-factor test was used to investigate the effect of ethanol volume fraction, liquid-solid ratio, extraction times and extracting time on the extraction rate of alisol B 23-acetate.The results were combined with Box-Benhnken design and response surface analysis to optimize the technology parameters for extraction process of Alismatis Rhizoma and obtain the optimal flash-type extraction technology under the following conditions： ethanol volume fraction 80%, liquid-solid ratio 12∶1, extraction 4 times, 114 s/time. Flash-type extraction technology of alisol B 23-acetate by response surface methodology is stable, time-saving, efficient, and with the advantages of room temperature extraction and no component damage, so it can be used for massive production.

Comparison on six active constituents between formula granule and traditional decoction of Rehmanniae Decoction of six ingredients / 中草药

Objective: To compare the content of chemical components in each drug in formula granule and traditional decoction of Rehmanniae Decoction with six ingredients. Methods: By using high performance liquid chromatography analysis and various chromatographic conditions, the contents were determined respectively, which were listed as follows: acteoside, loganin, paeonol, allantoin, pachymic acid and alisol B 23-acetate. Results: The contents of acteoside, loganin, paeonol, allantoin, pachymic acid and alisol B 23-acetate in traditional decoction were 304.5, 2 473.6, 3 135.1, 708.8, 5.9, and 104.4 μg/g, and they were 289.6, 3 685.7, 706.5, 714.2, 17.4, and 217.8 μg/g correspondingly in formula granule. The contents of acteoside and allantoin were basically the same between them; The contents of loganin, pachymic acid, and alisol B 23-acetate in formula granule were significantly higher than those in the traditional decoction; The content of paeonol in formula granule was significantly lower than that in the traditional decoction. Conclusion: The content difference of the chemical components is related to its chemical character between formula granule and traditional decoction.

Determination of cinnamic acid, cinnama ldehyde, paeonol, alisol B 23-acetate, atractylodes lancea and atractylodin in Yeming granules by RP-HPLC / 中国药学杂志

OBJECTIVE: To develop a RP-HPLC method for determination of cinnamic acid, cinnama ldehyde, paeonol, alisol B 23 acetate, atractylodes lancea, and atractylodin in Yeming granules. METHODS: The determination was performed on an Eclipse XDB-C18column (4.6 mm × 250 mm, 5 μm) at 30°C with a flow rate of 1.0 mL · min-1. The mobile phase was 0.3% phosphate-acetonitrile at gradient elution. The detection wavelength was set at 240 nm. RESULTS: The linear regression equations for cinnamic acid, cinnama ldehyde, paeonol, alisolB 23 acetate, atractylodes lancea, and atractylodin were ρ=-0.0055A + 0.0083 (r=0.9996), ρ=0.0255, 4+0.1336 (r=0.9999), ρ=0.0408 A +0.664 3 (r=0.999), ρ=0.003 3 4+0.0171(r=0.999 9), ρ=0.0163 A + 0.1075 (r=0.999 8), ρ=0.0229A + 0.0884 (r=0.9999), respectively. Good linearity was obtained over the ranges of 0.3096-2.322, 10.24-76.8, 37.36-280.2, 0.5128-3.846, 1.612-12.09 and 10.26-76.95 μg · mL-1, respectively. The recoveries were 100.1%, 99.8%, 100.7%, 99.3%, 100.4% and 100.5%, respectively. CONCLUSION: The method is simple and suitable to control the quality of Yeming granules. Copyright 2012 by the Chinese Pharmaceutical Association.

Transformation of alisol B 23-acetate in processing of Alisma orientalis / 中草药

Objective To study the transformation mechanism of triterpenes in processing of Alisma orientalis. Methods The triterpene transformations of A. orientalis pre and post-processing were comparatively analyzed by techniques of HPLC and Packed Column Supercritical Fluid Chromatography (SFC). Results In baked processing (70 ℃) of A. orientalis, little alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B.However, more alisol B 23-acetate was transformed into alisol A 24-acetate and alisol B, then both of them were further transformed into alisol A in processing under high temperature (160-200 ℃). Conclusion Transformation of alisol B 23-acetate has two routes when A. orientalis is processed under high temperature: For one, alisol B 23-acetate is rearranged into alisol A 24-acetate which could be deacetylated into alisol A; for the other; it is deacetylated into alisol B first, then transformed into alisol A.

Studies on Quality control for Rhizoma Alismatis / 中药新药与临床药理

Objective To establish a method for the quality control of Rhizoma Alismatis. Methods The samples of Rhizoma Alismatis were identified with TLC and the content of alisol B-23-acetate in Rhizoma Alismatis was determined by HPLC-ELSD. Results The spots of TLC chromatogram of Rhizoma Alismatis was clear. A good linearity was in the range of 4.4～110.0 ?g/mL of alisol B-23-acetate, r=0.999 2(n=5), and the average recovery rate was 98.98 %(n=9), RSD=1.3 %. Conclusion This method was simple, accurate and with a good reproducibility and can be used for the quality control of Rhizoma Alismatis.

Identification of Rhizoma Alismatis by HPLC Combined with Evaporation Light Scattering Determination Method / 中药新药与临床药理

Objective To establish a method for the identification of Rhizoma Alismatis. Methods HPLC combined with evaporation light scattering detector(ELSD) was applied. Chromatographic column was Macherey-Nagel C18(250 mm?4 mm,10 ?m),acetonitrile -water (4 ∶1) used as mobile phase,flow rate 0.5 mL/min,gas flow rate of ELSD 2.25 L/min and the temperature of drift tube at 70 ℃. Results In the extraction of the authentic Rhizoma Alismatis,both alisol A24-acetate and alisol B23-acetate could be estimated;in the extraction of Rhizoma Alismatis from the market,the alisol A24-acetate could be estimated only;and only alisol B23-acetate could be estimated in the extraction of the standard powder of Rhizoma Alismatis. Conclusion The method is efficient,practical and easy to operate for detecting and identifying Rhizoma Alismatis.

Optimum stir-bake with saltwater technology on Rhizoma Alismatis by multiple indexes orthogonal test / 中成药

AIM: To optimize and establish a stir-bake with saltwater technology on Rhizoma Alismatis by multiple indexes orthogonal test methods, providing technical index on standardization of stir-bake with saltwater on Rhizoma Alismatis. METHODS: The orthogonal test was employed to determine the effects of the three factors including the quantity of saltwater、temperature and time affiliated with three levels Alisol B23-acetate, alcohol extractive and properties. RESULTS: Temperature and time had notable effects on experimental results, but the quantity of saltwater had no effect on the results, the temperautre was at (100 ?C), time was 10 minutes an adding 2 kg salt to 10 kg water per 100 kg drugs. CONCLUSION: The research is meaningful to standardization of stir-bake with saltwater on Rhizoma Alismatis.