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OBJECTIVE@#Cassiae Semen (CS, Juemingzi in Chinese) has been used for thousands of years in ancient Chinese history for relieving constipation, improving liver function as well as preventing myopia. Here we aimed to elucidate the anti-steatosis effect and underlying mechanism of CS against non-alcoholic fatty liver disease (NAFLD).@*METHODS@#High-performance liquid chromatography (HPLC) was used to identify the major components of CS water extract. Mice were fed with a high-fat and sugar-water (HFSW) diet to induce hepatic steatosis and then treated with CS. The anti-NAFLD effect was determined by measuring serum biomarkers and histopathology staining. Additionally, the effects of CS on cell viability and lipid metabolism in oleic acid and palmitic acid (OAPA)-treated HepG2 cells were measured. The expression of essential genes and proteins involved in lipid metabolism and autophagy signalings were measured to uncover the underlying mechanism.@*RESULTS@#Five compounds, including aurantio-obtusin, rubrofusarin gentiobioside, cassiaside C, emodin and rhein were simultaneously identified in CS extract. CS not only improved the diet-induced hepatic steatosis in vivo, as indicated by decreased number and size of lipid droplets, hepatic and serum triglycerides (TG) levels, but also markedly attenuated the OAPA-induced lipid accumulation in hepatocytes. These lipid-lowering effects induced by CS were largely dependent on the inhibition of fatty acid synthase (FASN) and the activation of autophagy-related signaling, including AMP-activated protein kinase (AMPK), light chain 3-II (LC3-II)/ LC3-1 and autophagy-related gene5 (ATG5).@*CONCLUSION@#Our study suggested that CS effectively protected liver steatosis via decreasing FASN-related fatty acid synthesis and activating AMPK-mediated autophagy, which might become a promising therapeutic strategy for relieving NAFLD.
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The origins of 9 species of the Chinese medicinal materials in the 2015 edition of the Chinese pharmacopoeia(ChP) has revised in the 2020 edition of ChP. The revision is based on the investigation and textual research on the problems found after screening the original plants, animals or minerals of all the Chinese medicinal materials in the 2015 edition. Among them the Chinese names of Alismatis Rhizoma, Cassiae Semen, Coicis Semen, Corydalis Bungeanae Herba and Echinopsis Radix all do not match to the Latin scientific names, and also do not match the name of the actual medicinal origins. In addition, Alismatis Rhizoma has the omission of original plant. There is confusion about the Chinese name and the family name of the original insect of Cera Chinensis. The original mineral of Gypsum Fibrosum has the wrong group names. Alumstone and melanterite, the original mineral of Alumen and Melanteritum respectively, of which the group names are missing. To solve these problems, field survey and literature research were conducted on the medicinal materials and their origins. The source of these problems are explored. The correct origins and the Chinese names or Latin names are all determined according to the research results to the situation, in which the Chinese and Latin names of the original plants of the medicinal materials do not match. The correct family name and group name are obtained through textual research by taxonomy if the names are confused or mis-sing. The scientific evidence and correct results of revision in the 2020 edition of ChP are determined at last.
Subject(s)
Animals , China , Coix , Drugs, Chinese Herbal , Medicine, Chinese Traditional , RhizomeABSTRACT
The main chemical constituents of naphthopyrone reference extract( NRE) with definite content and relatively fixed chemical composition were analyzed and determined. Ultra-high performance liquid chromatography-LTQ-Orbitrap XL mass spectrometry and high performance liquid chromatography were used to systematically study NRE from the aspects of main chemical components and determination. The results showed that the chemical composition of naphthopyrone reference extract of Cassiae Semen was relatively fixed,and seven naphthalopyranones were identified. Cassiaside B_2,cassiaside C_2,rubrofusarin-6-O-β-D-gentiobioside and cassiaside C were the main chemical constituents of NRE,of which the determination and uncertainty results were( 11. 40+ 0. 26) %,( 11. 68+0. 24) %,( 16. 60+0. 22) %,( 28. 8+0. 48) %,respectively. This study contributed to the accurate evaluation of NRE and the foundation for the application of NRE in the quality control of Cassiae Semen,and provided a new idea for the replacement of single chemical reference substance by the reference extract of traditional Chinese medicine.
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Cassia , Chemistry , Certification , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Reference Standards , Mass Spectrometry , Plant Extracts , Reference Standards , Quality Control , Seeds , ChemistryABSTRACT
References and our previous experiment showed that the contents of glycosides were significantly decreased，while the contents of aglycones were significantly increased after processing of Cassiae Semen.It may be related to its glycosidases or the heating process. In order to investigate the reasons, high performance liquid chromatographic (HPLC) was used to study the effects of these two factors on contents of Cassiae Semen's main chemical components in processing. The results showed that glycoside hydrolases was present in Cassiae Semen and could rapidly hydrolyze glycosides from Cassiae Semen into aglycones in suitable temperature with sufficient water.However，it didn't show effect on contents change of main constituents in the procedure of Cassiae Semen processing.The reason for content decrease of glycosides and content increase of aglycones in processed Cassiae Semen was glycoside bond cracking to produce corresponding aglycone at high temperature.This study further provides basis for further revealing of the processing mechanism of Cassiae Semen.
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Objective: To explore the lipid regulating effect of fermentation product of Crataegi Fructus, Alismatis Rhizoma, and Cassiae Semen on hyperlipidemic rats. Methods: SD rats were fed with high fat diet and established as hyperlipidemia animal model, the subjects were divided into six groups: control group, model group, positive control group, fermentation product prevention group, red yeast rice group, and fermentation product treatment group. After four weeks continuous oral administration, the effect of different medicine samples on serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) of hyperlipidemic rats were observed to assess the regulation effect. Results: Four weeks' continuous oral administration could regulate the TC, TG, LDL-C levels significantly lower (P < 0.001) and HDL-C level significantly increased (P < 0.05) in the prevention group compared with the model group. After modeling, continuous administration for four weeks, compared with the levels before giving medicines, TC, TG, LDL-C levels of positive control group and fermentation product treatment group were significantly regulated lower (P < 0.05, 0.01) and HDL-C level significantly increased (P < 0.05, 0.01). while the only TC and LDL-C levels in red yeast rice group showed significant effect. Conclusion: Product of lipid-lowering traditional Chinese medicines (TCMs) treated by solid-state fermentation with Monascus purpureus could effectively inhibit the formation of foodborne hyperlipidemia, as well as control and regulate hyperlipidemia, and synergistic effect also appeared between ingredients from TCMs and lovastatin.
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Objective To establish a UPLC fingerprint of Cassiae Semen from two species which is expected to provide standard for the quality control and evaluation. Methods Chromatography conditions were Agilent ZORBAX EP C18 (100 mm × 3.0 mm, 1.8 μm) column with gradient mobile phase of acetonitrile-0.01% formic acid solution. UV detection wavelength was 285 nm, the flow rate was 0.4 mL/min, the column temperature was 30 ℃ and the sample quantity was 1 μL. Results The common mode of UPLC fingerprint of Cassiae Semen was established firstly. There were 20 common peaks had been calibrated by similarity evaluation. Seven peaks had been identified by comparing retention time with UV spectrum analysis. The similarity evaluation of 20 batches samples of Cassiae Semen was more than 0.9. Twenty batches of Cassiae Semen could be divided into four grades when the sum of squared Euclidean distance is 5 become the result of Cluster Analysis. Principal component analysis got three principal components through dimension reduction. The results of principal component scores agree with the clustering analysis. By fitting the load factor model of the first principal component, characteristic peak such as rubrofusarin-6-O-β-gentiobioside were filtered to appraise Cassiae Semen's quality. Conclusion The contents of same chemical compositions between Cassia obtusifolia and C. tora are different. UPLC fingerprint can be used in Cassiae Semen's quality control and evaluation.
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In this study, the authors reviewed domestic and foreign literatures, conducted the textual research on origin and development of Cassia Semen, studied records in ancient books and ancient and modern literatures, clinical adverse reactions and relevant experimental studies in recent years, and summarized the clinical features and influencing factors related to the safety of Cassiae Semen. According to the findings,Cassia Semen's safety risks are mainly liver and kidney system damages, with the main clinical features of fatigue, anorexia, disgusting of oil, yellow urine and gray stool; digestive system injury, with the main clinical features of diarrhea, abdominal distension, nausea and loose stool; reproductive system damage, with the main clinical features of vaginal bleeding. Allergic reactions and clinical adverse events, with the main clinical features for numb mouth, itching skin, nausea and vomiting, diarrhea, wheezing and lip cyanosis were also reported. The toxicological studies on toxic components of Cassiae Semen obtusifolia were carried out through acute toxicity test, subacute toxicity test, subchronic toxicity test and chronic toxicity test. Risk factors might include patients, compatibility and physicians. Physicians should strictly abide by the medication requirements in the Pharmacopoeia, pay attention to rational compatibility, appropriate dosage,correct usage and appropriate processing, control the dosage below 15 g to avoid excessive intake, strictly control the course of treatment to avoid accumulated poisoning caused by long-term administration. At the same time, clinicians should pay attention to the latest research progress, update the knowledge structure, quickly find the latest and useful materials from clinical practice, scientific research and drug information and other literatures, make evaluation and judgment for the materials, establish a traditional Chinese medicine intelligence information library, and strengthen the control over adverse effects with a pre-warning consciousness. The authors suggested standardizing clinical medication of Cassiae Semen, and avoiding misuse or excessive use; clinicians should prescribe it in strict accordance with there commended usage and dosage in the Pharmacopoeia, and focus on the safety signal accumulation in clinic, while strengthening studies for toxic substance basis and toxicity mechanism, in order to give full play to Cassiae Semen's clinical efficacy and reduce its adverse reactions.
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Objective: To establish a quantitative analysis of multi-components by single mark (QMSA) for determining the contents of four anthraquinones (aurantio-obtusin, emodin, rhubarb, and physcion) from Cassiae Semen, and verify the feasibility of this method in quality control. Methods: The medicinal material of Cassiae Semen was used as the research object, emodin was used as reference, two kinds of correction methods were used to establish the relative correction factor (fk/s) of aurantio-obtusin, emodin, and physcion. The amount of each component was calculated, at the same time using the external standard method for determination of the three components, comparing the difference between calculated values of QMSA and measured value to validate the feasibility and accuracy of QMSA. Results: The relative correction factors of aurantio-obtusin, emodin, and physcion were established, without significant difference between the calculated values by QMSA and measured values. Conclusion: The experimental QMSA method can accurately determine the contents of the four anthraquinones in Cassiae Semen, and can be used in the multi-index evaluation in Cassiae Semen anthraquinones constituents.
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Objective: To obtain a new lipid-lowering product that combines the advantages of lipid-lowering Chinese materia medica (CMM) and statin drugs. Solid-state fermentation was studied using medicinal and edible CMM as media components, Lovastatin yielding monascus screened in our laboratory was used as strain and Lovastatin yielding was used as index. Methods: Solid-state fermentation was optimized by single factor and orthogonal test, qualitative and quantitative analysis was conducted of the product via TLC and HPLC. Results: The optimal fermentation conditions contained 100 g/solid medium in 500 mL flask bottling capacity, material thickness of 2.5 cm, 10 mL liquid seed of 48 h, 10% inoculation volume, incubating at 30℃, breaking up the medium on day 2, adding 24% sterile water on day 3, cooling to 25℃ and culturing for 18 d in total, and Lovastatin yielding was up to 5.127 mg/g. Fermentation product of CMM contained more components compared with that without medicines, Lovastatin yielding increased by 42.27%, but γ-aminobutyric acid yielding decreased by 17.89%. Contents of main active ingredients ursolic acid, 2,3-acetyl alisol-B, chrysophanol, and physcion were increased by 232.7%, 173.7%, 767.6%, and 888.4%. Conclusion: Active ingredients of hawthorn, alisma, and cassia are released into the products after fermentation and contents of lipid-lowering active ingredients were improved significantly, new active ingredients are also noticed. Therefore, the fermentation process of lipid-lowering medicines obtained in this experimental study has some practical values.
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To study the effect of aqueous extract of Cassiae Semen on the activity, mRNA and protein expressions of cytochrome P450(CYP450) system in rat liver microsomes, microsomes of rat liver were prepared after the oral administration with aqueous extract of Cassiae Semen for 14 days. The enzyme activity was quantified by Cocktail method. Meanwhile, the mRNA and protein expressions of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1 in the livers were detected by RT-PCR and Western blot. The result of this experiment was that aqueous extract of Cassiae Semen obviously induced the enzyme activities of CYP1A2, CYP2B1, CYP2C11, CYP2D2, CYP2E1 and CYP3A1. Low dose of aqueous extract of Cassiae Semen significantly reduced the activity of CYP2D2, but the activity of CYP2D2 was significantly induced by middle dose and high dose of aqueous extract of Cassiae Semen. These subtypes were increased in a dose-dependent manner except for CYP3A1. The mRNA levels of CYP1A2, CYP2C11, CYP2D2 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant effect in CYP2B1 and CYP3A1 mRNA expressions. The protein levels of CYP2C11 and CYP2E1 were also induced in rats treated with aqueous extract of Cassiae Semen, but with no significant difference. Since the enzyme activity, mRNA and protein expressions of CYP450, particularly CYP2C11and2E1subtypes, were induced or inhibited by aqueous extract of Cassiae Semen to varing degrees, suggesting the potential drug-drug interactions should be concerned.
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RESULTS: Rubrofusarin gentiobioside and isorubrofusarin gentiobioside could convert in water and ammonia at a certain temperature, but the reaction could not happen in acid solvent. On the other hand, solvent and temperature could affect the stability of the two compounds.