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1.
Article in Chinese | WPRIM | ID: wpr-907682

ABSTRACT

Objective:To establish the HPLC fingerprint method for assessing the quality of Moutan Cortex, and to determine the contents of paeonol, paeoniflorin, gallic acid, hydroxyl-paeoniflorin and benzoyl-paeoniflorin of Moutan Cortex in different growth period. Methods:Diamonsil Plus C18 column (250 mm × 4.6 mm, 5 μm) was used with the mobile phase comprising acetonitrile-0.05% formic acid solution and the flow rate of 1.0 ml/min with gradient elution manner. The detected wavelength was 230 nm for paeoniflorin and benzoyl-paeoniflorin, 267 nm for gallic acid, 258 nm for hydroxyl-paeoniflorin and 274 nm for paeonol with temperature column of 25 ℃. Then putting chromatograms into Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Medica (2012A) to evaluate the similarity of Moutan Cortex in different growth period; then putting peak area data into SPSS software for cluster analysis and the clustering effect was determined. Results:The HPLC fingerprints established with this method has 23 shared peaks and 5 of them were identified, namely, paeonol, paeoniflorin, gallic acid, hydroxyl-paeoniflorin and benzoylpaeoniflorin. The similarity of Moutan Cortex in different years was between 0.850-0.991. This method has good linear relation ( r≥0.999 5), RSDs of precision, stability tests and reproducibility were lower than 1.6% ( n=6). Different growth periods of Moutan Cortex have obvious influence on the concentration of five compounds. Conclusion:This method is useful to evaluate and discriminate Moutan Cortex at different growth periods so as toprovide scientific reference on the harvest,industrialization and evaluation of Moutan Cortex.

2.
China Pharmacy ; (12): 2846-2853, 2021.
Article in Chinese | WPRIM | ID: wpr-906650

ABSTRACT

OBJECTIVE:To study the improvement effects of paeon iflorin(PF)on myocardial injury in type 2 diabetes mellitus(T2DM)model rats and its mechanism. METHODS :The experiment was set up in the normal group ,model group , positive control group (metformin 90 mg/kg),PF high-dose ,medium-dose and low-dose groups (90,60,30 mg/kg),with 8 rats in each group. Except for normal group ,other groups were given high-glucose and high-fat diet and intraperitoneal injection of streptozotocin (30 mg/kg) to induce T 2DM model. After modeling , administration groups were given relevant medicine intragastrically,normal group and model group were given constant volume of normal saline intragastrically ,once a day ,for consecutive 4 weeks. The body weight ,fasting blood glucose and oral glucose tolerance were measured ;serum levels of glycosylated serum protein (GSP),total cholesterol (TC),triacylglycerol(TG),glutathione peroxidase (GSH-Px),superoxide dismutase(SOD),malondialdehyde(MDA),creatine kinase isoenzyme-MB (CK-MB) and troponin Ⅰ (cTn Ⅰ) were determined. The pathomorphological changes of myocardium were observed. The apoptosis index of rat cardiomyocytes was ( detected. The protein expression of B-cell lymphoma 2 (Bcl-2),Bcl-2 related X protein (Bax)and caspase- 3 in rat myocardium were detected by immunohistochemistry and Western blot. RE SULTS:Compared with normal group ,the body weight ,serum levels of GSH-Px and SOD ,protein expression of Bcl- 2 in myocardium were decreased significantly in model group(P<0.01);while fasting blood glucose ,area under blood glucose curve ,serum levels of biochemical indexes (GSP,TC, TG,MDA,CK-MB,cTnⅠ),cardiomyocyte apoptosis index ,protein expression of Bax and caspase- 3 in myocardium were increased significantly (P<0.05 or P<0.01). The arrangement of myocardium was relatively irregular ,and some muscle fibers were broken. Compared with model group ,except for body weight ,serum levels of SOD and MDA ,the protein expression of Bax in myocardium in PF low-dose group , above indexes of PF groups were reversed significantly (P<0.05 or P<0.01). CONCLUSIONS:PF can regulate glycolipid metabolism ,enhance antioxidant ability ,inhibit cardiomyocyte apoptosis and improve myocardial injury in T 2DM model rats ;the mechanism may be associated with increasing the protein expression of Bcl- 2 and down-regulating the protein expression of Bax and caspase- 3 in myocardium.

3.
Article in Chinese | WPRIM | ID: wpr-906038

ABSTRACT

Paeoniae Radix Rubra is a traditional Chinese medicine commonly used in clinical practice, it is mostly wild and widely distributed in different areas of China. In addition, the plant of Paeoniae Radix Rubra also has ornamental value. Modern phytochemical researches showed that the chemical constituents of Paeoniae Radix Rubra were complex. Up to now, more than 300 chemical constituents have been found, mainly including monoterpene glycosides, triterpenoids, flavonoids, tannins, phenolic acids, saccharides, steroids, volatile oils and so on. Among them, the content of monoterpene glycosides was the highest, and the types of volatile oil were the most. Paeoniae Radix Rubra has a wide range of pharmacological effects, exerting different curative effects in multiple systems such as blood, cardiovascular, nervous and digestive system. It can protect myocardial cells and nerve cells, stabilize microcirculation, anti-endotoxin, anti-atherosclerosis, reduce pulmonary hypertension, anti-depression, protect liver, anti-gastric ulcer, anti-tumor, slow down aging, treat Parkinson's syndrome and diabetes and its complications, anti-radiation, anti-inflammatory, anti-virus and so on. Through reviewing the literature on chemical constituents and pharmacological effects of Paeoniae Radix Rubra, it was found that total glycosides and monomers such as paeoniflorin, albiflorin, benzoylpaeoniflorin and gallic acid may be the main active components of Paeoniae Radix Rubra. At present, the research on Paeoniae Radix Rubra mainly focused on monoterpene glycosides, while the research on flavonoids and volatile oil in Paeoniae Radix Rubra was less. It is suggested that research on these two components should be strengthened in the future.

4.
Article in Chinese | WPRIM | ID: wpr-888120

ABSTRACT

The study aims to investigate the effect of the compatibility of paeonol and paeoniflorin(hereinafter referred to as the compatibility) on the expression of myocardial proteins in rats with myocardial ischemia injury and explore the underlying mechanism of the compatibility against myocardial ischemia injury. First, the acute myocardial infarction rat model was established by ligation of the anterior descending branch of the left coronary artery. The model rats were given(ig) paeonol and paeoniflorin. Then protein samples were collected from rat cardiac tissue and quantified by tandem mass tags(TMT) to explore the differential proteins after drug intervention. The experimental results showed that differential proteins mainly involved phagocytosis engulfment, extracellular space, and antigen binding, as well as Kyoto encyclopedia of genes and genomes(KEGG) pathways of complement and coagulation cascades, syste-mic lupus erythematosus, and ribosome. In this study, the target proteins and related signaling pathways identified by differential proteomics may be the biological basis of the compatibility against myocardial ischemia injury in rats.


Subject(s)
Acetophenones , Animals , Glucosides , Monoterpenes , Myocardial Ischemia/genetics , Myocardial Reperfusion Injury , Proteomics , Rats , Rats, Sprague-Dawley
5.
Acta Pharmaceutica Sinica ; (12): 1811-1819, 2021.
Article in Chinese | WPRIM | ID: wpr-887029

ABSTRACT

The current study was designed to evaluate the modulatory effects of paeoniflorin on the dysregulated gut microbiota as well as the disturbed fecal bile acids (BAs) in colitis mice. After approved by Xi'an Jiaotong University Ethics Committees (Approval No. XJTU2019-679), the animals were randomly distributed into the control (Con), colitis, low dose paeoniflorin (PF-L, 25 mg·kg-1·d-1), high dose paeoniflorin (PF-H, 50 mg·kg-1·d-1) and 5-aminosalicylic acid (5-ASA, 50 mg·kg-1·d-1) groups. Colitis was induced by administering 3% (w/v) DSS in drinking water for 7 days. Paeoniflorin and 5-ASA were dissolved in water and administered to the appropriate groups by oral gavage over the 7-day period. The mice were monitored daily, and the disease activity index (DAI) comprising of body weight loss, stool consistency and gross blood was measured. The pathological changes of colon were evaluated by HE staining; the levels of inflammatory cytokines in colonic tissue were determined by ELISA; the gut permeability was measured by FITC-dextran. Microbiota analysis based on 16S rDNA and targeted metabolomics for BAs were used to evaluate the composition of gut microbiota and fecal BAs pool. The results showed that administration of paeoniflorin markedly alleviated the inflammatory response and intestinal barrier dysfunction in DSS-induced colitis. Importantly, these ameliorative effects of paeoniflorin were accompanied by the improvements of disturbed composition of gut microbiota and the dysmetabolism of bile acids in feces. Finally, we performed Spearman's correlation analysis between the fecal BAs and gut microbiota genera, and found that Lactobacillus has a strong positive correlation with DCA and LCA which were reported to confer the beneficial effects of maintaining intestinal homeostasis. Taken together, paeoniflorin might improve the intestinal homeostasis in colitis mice via modulating gut microbiota and fecal BAs metabolism.

6.
Article in English | WPRIM | ID: wpr-881058

ABSTRACT

This study was to investigate the protective effect of paeoniflorin (PF) on hydrogen peroxide-induced injury. Firstly, "SMILES" of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets. Injury-related molecules were obtained from GeenCards database, and the predicted targets of PF for injury treatment were selected by Wayne diagram. For mechanism analysis, the protein-protein interactions were constructed by String, and the KEGG analysis was conducted in Webgestalt. Then, cell viability and cytotoxicity assay were established by CCK8 assay. Also, the experimental cells were allocated to control, model (200 μmol·L

7.
Article in Chinese | WPRIM | ID: wpr-846626

ABSTRACT

Objective: To establish chemical fingerprint and multi-components determination of 15 batches of Taohong Siwu Decoction (TSD), and provide reference for the improvement of its quality control. Methods: The separation was performed on Thermo Hypersil Gold C18 column (250 mm × 4.6 mm, 5 μm) for gradient elution with methanol-0.1% phosphoric acid aqueous solution, flow rate 1.0 mL/min, column temperature 30 ℃, and detection wavelength 225 nm. The HPLC fingerprint was established and evaluated by the similarity evaluation system of TCM (version 2012A), and the difference of chemical information between 15 batches of different samples was evaluated by cluster analysis. Furthermore, the content of the nine active components in the sample was determined by HPLC multi-component wavelength switching method, with the partial least squares-discriminant analysis (PLS- DA) by SIMCA 14.1 software to find significant components of the quality between the batches. Results: The HPLC fingerprint of 15 batches of TSD was established. The similarity was greater than 0.96, and 35 common peaks were identified as gallic acid, chlorogenic acid, amygdalin, albiflorin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, senkyunolide I, benzoylpaeoniflorin and ligustilide (corresponding to peaks 2, 8, 9, 13, 14, 15, 16, 25, 31, and 32). The linearity relationships of gallic acid, 5-hydroxymethylfurfural, chlorogenic acid, albiflorin, hydroxysafflor yellow A, paeoniflorin, ferulic acid, verbascoside, and senkyunolide I (r ≥ 0.999 6) were good. The results of content determination respectively were 187.5-344.4, 6.2-154.8, 413.2-459.2, 507.5-923.5, 873.8-1 202.0, 2 122.3-2 782.9, 59.2-121.3, 6.4-26.9, and 38.9-79.6 μg/g, respectively, including higher content of paeoniflorin, hydroxysafflor yellow A, and albiflorin. Furthermore, 15 batches of samples from different origins were classified into three categories. Using PLS-DA analysis, the content determination result showed that paeoniflorin, albiflorin, hydroxysafflor yellow A, and 5-hydroxymethylfurfural were the four components that affected the quality of different batches of TSD. Conclusion: HPLC fingerprint combined with multi-components determination is suitable for quality control and evaluation of TSD preparation.

8.
Article in Chinese | WPRIM | ID: wpr-846566

ABSTRACT

Objective: To investigate the therapeutic effect and potential mechanism of Zhenwu Decoction (ZWD) on chronic heart failure (CHF) rats. Methods: HPLC fingerprint of ZWD was established. All male SD rats were randomly divided into the sham operation group, the model group, the low, medium and high dose ZWD group (2.187 5, 4.375, and 8.75 g/kg) and the captopril group (10 mg/kg). Except for the sham operation group, the rest of rats were all established into the CHF model rats by ligating the left anterior descending branch of the coronary artery, after 8 weeks, all rats were ig administration for 4 weeks. The hemodynamic, viscera index, HE dyeing test were conducted at the end of experiments. Serum angiotensin II (Ang II), aldosterone (ALD), nuclear factor kappa B (NF-κB), amino terminal brain natriuretic peptide (NT-proBNP), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) were determinated by ELISA, and myocardial NF-κB protein expression was detected by Western blotting. Results: Higenamine, paeoniflorin, atractylenolide III, 6-gingerol and dehydrotumulosic acid, the five constituents of Zhenwu Decoction, were identified by HPLC chart. Compared with the model group, the administration of the ZWD significantly improved the hemodynamic parameters (P < 0.05), reduced the organ index (P < 0.05) and improved myocardial injury, reduced the serum Ang II, ALD, NF-κB, NT-proBNP, TNF-α and IL-6 levels and the myocardial NF-κB protein expression (P < 0.05). Conclusion: HPLC results provided an evidence for the quality control and pharmacodynamic substance of ZWD. ZWD can ameliorate CHF, which may be related to the inhibition of renin- angiotensin-aldosterone system (RAAS)/NF-κB/inflammatory factor cascade reaction.

9.
Article in Chinese | WPRIM | ID: wpr-846412

ABSTRACT

Objective: To explore the potential Q-marker of Paeoniae Radix Alba in Danggui Sini Decoction based on fingerprint and network pharmacology. Methods: The fingerprints of Paeoniae Radix Alba decoction and Danggui Sini Decoction were established, and the law of components transfer was also defined. The "compounds-targets-pathways" network was then established to predict the potential Q-marker of Paeoniae Radix Alba through the network pharmacology. Results: The fingerprints of 15 batches of Paeoniae Radix Alba decoction and 15 batches of Danggui Sini Decoction were established, and the five chromatographic peaks were identified, they were gallic acid, albiflorin, paeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, and benzoyl paeoniflorin. Through the network pharmacology analysis, the potential two active components, eight core targets and 13 key pathways were screened out, which indicated that paeoniflorin and albiflorin were preliminarily predicted to the potential Q-marker of Paeoniae Radix Alba. Conclusion: The analysis and prediction of the Q-marker in this study can provide a reference for the whole control of the Paeoniae Radix Alba quality, which can also provide the basis for the further research on the efficacy-related substance and mechanism of Paeoniae Radix Alba.

10.
Article in Chinese | WPRIM | ID: wpr-846325

ABSTRACT

Objective: Using ultra-high performance liquid chromatography with diode array detection (UHPLC-DAD) and desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) to analyze 15 batches of Shaoyao Gancao Decoction (SGD) substance benchmark and lyophilized powder in order to investigate the advantages of DESI-MSI in quality control of famous classical formulas. Methods: Taking SGD as the research model, fingerprints of the substance benchmark were established by UHPLC-DAD, and the content of index components (paeoniflorin, liquiritin, glycyrrhizic acid) and the yield of dry extract were also investigated. Meanwhile, as the research carrier, the lyophilized powder corresponding to SGD was dissolved in methanol and dotted on qualitative filter paper with quantitative capillary, and fixed it on the slide to make samples. The samples were analyzed on a DESI-MSI system in positive and negative ion mode with methanol-formic acid (1 000:1, flow rate of 3 μL/min) as spray solvent, N2 as spray gas (pressure of 0.5 MPa). The scanning range was m/z 100-1 200, the spatial resolution was 300 μm, the ion source temperature was 120 ℃. Results: DESI-MSI can detect not only the index components of paeoniflorin, liquiritin, glycyrrhizic acid, but also the common peaks of albiflorin. At the same time, DESI-MSI could detect 11 other components from Glycyrrhizae Radix et Rhizoma and Paeoniae Radix Alba, such as licoricesaponin G2, licoricesaponin J2, gallic acid, citric acid, p-hydroxybenzoic acid, and present their relative content visually. The qualitative analysis ability of DESI-MSI was much better than UHPLC-DAD. Conclusion: DESI-MSI can be used as the quality control method for substance benchmark and lyophilized powder and dispensing granules of classical famous formulas with advantages of high sensitivity, strong analytical ability, no complex sample pretreatment, qualitative and relative content analysis of complex samples without reference substance.

11.
Article in Chinese | WPRIM | ID: wpr-846296

ABSTRACT

Objective: To develop and validate an high performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC-ESI-MS/MS) method for simultaneously qualitative and quantitative determination of 13 major bioactive components (ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a, and saikosaponin d) in Jiawei Zuojin Pills (JZP). Methods: The chromatographic separation was performed on a Thermo Syncronis C18 column (100 mm × 4.6 mm, 3.0 μm) with a gradient elution of methanol and 0.02 mol/L ammonium acetate in water at a flow rate of 0.35 mL/min, and the injection volume was 10 μL. The 13 major bioactive components were detected using an electrospray ionization source in ESI+ and ESI- ionization mode, quantified by multiple reaction monitor (MRM) scanning at the same time. Results: The linear ranges of ferulic acid, costunolide, baicalin, paeoniflorin, tetrahydropalmatine, rutecarpine, berberine, palmatine, evodiamine, naringin, hesperidin, saikosaponin a and saikosaponin d were 2-80 μg/mL (r = 0.999 2), 0.5-20.0 μg/mL (r = 0.999 1), 3.5-140.0 μg/mL (r = 0.999 8), 1-40 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 2), 3-120 μg/mL (r = 0.999 7), 2.5-100.0 μg/mL (r = 0.999 5), 0.5-20.0 μg/mL (r = 0.999 3), 0.5-20.0 μg/mL (r = 0.999 1), 1-40 μg/mL (r = 0.999 1), 0.3-12.0 μg/mL (r = 0.999 2), 0.3-12.0 μg/mL (r = 0.999 2), and the average recoveries were 99.5% (RSD = 4.11%), 98.9% (RSD = 4.88%), 100.2% (RSD = 1.08%), 99.2% (RSD = 3.23%), 99.5% (RSD = 4.13%), 99.7% (RSD = 3.23%), 98.6% (RSD = 2.78%), 99.9% (RSD = 3.12%), 101.3% (RSD = 4.53%), 98.7% (RSD = 3.43%), 99.8% (RSD = 3.58%), 97.9% (RSD = 5.22%), and 101.3% (RSD = 5.13%), respectively. The contents of 12 batches of the 13 major bioactive components were 0.324-0.383, 0.051-0.072, 3.225-3.466, 0.154-0.198, 0.015-0.062, 0.144-0.199, 2.145-2.982, 0.441-0.953, 0.032-0.099, 0.062-0.089, 0.111-0.178, 0.012-0.065, 0.011-0.069 mg/g, respectively. Conclusion: The developed method is simple, specific, and sensitive, and it can be applied for the determination of 13 major bioactive components and the quality control of JZP.

12.
Article in Chinese | WPRIM | ID: wpr-846172

ABSTRACT

Objective: To establish the HPLC fingerprint and simultaneously determinate nine components of the standard decoction of Wenjing Decoction, so as to provide reference for the quality control of Wenjing Decoction of classical prescriptions. Methods: Fingerprints of 15 batches of the standard decoction of Wenjing Decoction were determined by HPLC-PDA, and the control fingerprint was established. All samples were analyzed by Kromasil C18 chromatographic column (250 mm × 4.6 mm, 5 μm) maintained at 25 ℃, and eluted with acetonitrile-0.1% phosphoric acid at the flow rate of 0.8 mL/min, and the detection wavelength was 220, 280, 320 and 380 nm respectively. Combined with cluster analysis (CA), principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA), the quality of 15 batches of Wenjing Decoction was analyzed. At the same time, the contents of nine active components were determined. Results: The similarity of 15 batches of standard decoction of Wenjing Decoction was between 0.902 and 0.992, and a total of 18 common peaks were identified and nine of them (2-gallic acid, 5-paeoniflorin, 7-liquiritin, 8-ferulic acid, 9-isoliquiritin apioside, 11-isoliquiritin, 14-cinnamaldehyde, 15-ammonium glycyrrhetate, 16-paeonol) were quantitative analyzed. CA, PCA and PLS-DA were used to classify the 15 batches of samples into two groups. The results of quantitative analysis were good, and the recovery rate of nine components was 94.91%-108.16%. The content of gallic acid, paeoniflorin, iquiritin, ferulic acid, isoliquiritin apioside, isoliquiritin, cinnamaldehyde, ammonium glycyrrhetate, paeonol in 15 batches of samples were in the range of 10.7-31.3, 95.8-228.4, 18.6-62.4, 3.3-8.3, 4.8-18.7, 2.8-10.6, 13.7-108.2, 83.9-292.3, and 31.1-125.5 mg/g, respectively. Conclusion: The HPLC fingerprint combined with the simultaneous determination of multicomponent analysis method eatablished in this experiment are stable and reliable, which can provide the theoretical guidance for the quality evaluation of Wenjing Decoction and its compound preparations.

13.
Article in Chinese | WPRIM | ID: wpr-846146

ABSTRACT

Objective: To study the content variation and chemical composition of Siwu Decoction between mixed decoction and single decoction comprehensively, and then explore variation rule of Siwu Decoction by different decocting methods based on material basis. Methods: Components of Siwu Decoction were identified by LC-MS/MS and an UPLC wavelength switching method for simultaneously determining the contents of multiple compounds in Siwu Decoction was established based on the idea of TCM chemistry holography. The mixed and single decoction samples were prepared and tested. Experimental data was compared to analyze material basis differences and variation rule of Siwu Decoction by different decocting methods. Results: A total of 72 compounds were identified and assigned, 18 compounds were quantitative detected and all of 18 analytes showed good linearity (R2 ≥ 0.999) within the test range. The relative standard deviations of the precision, repeatability and stability were not exceeding 2.0%, and the recoveries were in the range of 97%-105%. Analysis of Siwu Decoction samples showed dissolution of ligustilide, 3-n- butylphthalide, catechin, gallic acid and paeoniflorin was affected by the change of solvent volume and dissolution of aucubin, catechin, oxypaeoniflorin, paeoniflorin and acteoside were higher in mixed decoction than single decoction obviously. Compared to single decoction, the kinds of compounds in mixed decoction did not change significantly but the content showed notable variety. Conclusion: Through the study of chemistry holography, the composition and content of compounds in TCM mixed decoction and herbs single decoction can be compared and analyzed comprehensively to provide a new perspective for the study on the rule of TCM decoction and dissolution. TCM chemistry holography study may become a useful exploration of the TCM quality study.

14.
Article in Chinese | WPRIM | ID: wpr-846117

ABSTRACT

Objective: Through network pharmacology, the network relationship between the active component of Sanqi Mixture, the target of hepatic ischemia- reperfusion injury(HIRI), and biological pathway was constructed to explore the key target and mechanism of effect of Sanqi Mixture on HIRI. Method: Through literature research at home and abroad, Traditional Chinese Medicine Systems Pharmacology (TCMSP) platform, Pharm Mapper, Swiss Target Prediction and other servers, oral availability (OB) and drug-likeness (DL) were selected as the limited conditions to collect the relevant targets for Sanqi Mixture for intervention in HIRI. The OMIM database was used to screen and collate HIRI related genes and protein targets. Excel table was used to merge and sort the intersection between disease and targets through Cytoscape3.7.2 software plug-ins Network Analyzer, with topological parameters (degree) ≥ 5 (average degrees of freedom 4.5) for the filter to find the core targets; And the intersection targets were imported to the server STRING, and with Confidence Score of 0.85 or higher for the filter conditions to build the core protein interactions (Hub-PPI) network. The intersection target was introduced into FunRich 3.0 software for biological process and biological pathway analysis, and Cytoscape3.7.2 was used to construct the network of "traditional Chinese medicine-active ingredient-HIRI target-biological pathway". Result: Sanqi mixture could reduce the expression of Aspartate aminotransferase (AST) and glutamate transaminase (ALT) in HIRI mice (P < 0.01). After screening, 45 active components of Sanqi Mixture were obtained, corresponding to 3 273 targets, and the main compounds included ursolic acid, oleanolic acid, brucine, quercetin, ginsenoside F2, paeoniflorin, etc. Among the 196 targets obtained by HIRI, 46 targets were intersected with components, including 11-β-hydroxysteroid dehydrogenase (HSD11B1), adenosine receptor A3 (ADORA3), cyclooxygenase 2 (PTGS2), adenosine receptor A1 (ADORA1), protein kinase C-ε (PKC), etc. With the STRING server setting the qualified condition of Confidence Score ≥ 0.85, the PPI network with high Confidence was obtained and clustered into three categories through cluster processing. Five biological processes including protein metabolism, signal transduction, negative regulation of enzyme activity, inflammatory response and transmembrane receptor protein tyrosine kinase signal pathway were analyzed by FunRich software (P < 0.05). 16 biological pathways including integrin-linked kinase signal, TNF receptor signaling pathway, P38 mitogen-activated protein kinase signaling pathway, and TRAIL signaling pathway (P < 0.01). Conclusion: It is preliminarily discussed that Sanqi Mixture intervenes HIRI through the interaction of multiple components and multiple targets, as well as the regulation of multiple biological pathways and biological processes. However, the key core targets and the specific regulation mechanism still need further experimental verification.

15.
Article in Chinese | WPRIM | ID: wpr-846107

ABSTRACT

Objective: To establish the UPLC specific chromatogram and HPLC content determination methods of multi-index components about the material reference of classical Huaganjian and build its quality control system. Methods: According to the ancient books and combining with the previously inspected process, 18 batches of Huaganjian material reference from different origins were prepared. The specific chromatogram was established by using UPLC. Similarity was calculated by using TCM Chromatographic Fingerprint Similarity Evaluation Software (2012). Combining with orthogonal partial least squares discriminant analysis, we excavated the main components that affected the quality of Huaganjian material reference from different batches and origins. Three of these index components (paeoniflorin, hesperidin, paeonol) from prescription sovereign drug, minister drug, and assistant drug were selected and used as indicators for content determination of Huaganjian material reference. HPLC content determination methods were established and the content of 18 batches of samples was determined respectively. Results: The similarity of the specific chromatogram was ≥ 0.989. Thirty-three common peaks were calibrated, and eight common peaks were identified by chemical composition (gallic acid, geniposide, paeoniflorin, hesperidin, didymin, paeonol, sinensetin, and 3,5,6,7,8,3',4'- heptamethoxyflavone). Nine index components that affected the stability between batches were found out (Peak 31, 20, 11, 13, 22, 33, 21, 29, 1). Paeoniflorin, hesperidin, and paeonol were selected as content determination indicators. The content range of these components in material reference was 1.28%-1.95% paeoniflorin, 0.91%-1.02% hesperidin, 0.48%-0.57% paeonol. Conclusion: The quality control method of the material reference of classic prescription Huaganjian was established preliminarily through the UPLC specific chromatogram and HPLC content determination of index components. This method was rapid, simple, feasible, reproducible, stable and could provide a theoretical basis for the subsequent development and quality control of Huaganjian preparations.

16.
Article in Chinese | WPRIM | ID: wpr-846046

ABSTRACT

Objective: To establish a UPLC method to simultaneously determine 10 active ingredients in Yiganning Granules (YG) and provide scientific basis for the quality control, evaluation and standard revision of YG preparations. Methods: A UPLC method was used with an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm). The mobile phase was acetonitrile-mehanol-0.15% phosphoric acid solution with gradient elution. The flow rate was 0.3 mL/min. The column temperature was 45 ℃. The injection volume was 2 μL. Results: Ten active ingredients (chlorogenic acid, atractylenolide I, paeoniflorin, calycosin 7-O-β-D- glucopyranoside, stilbene glycoside, caffeic acid, toosendanin, kaempferol, paeonol and tanshinone ⅡA) in YG were simultaneously determined. The linearity was good (r ≥ 0.999 0), the limit of detection and quantification were 0.006-0.017 μg/mL and 0.017-0.510 μg/mL. The average recoveries were 98.8%-102.5% with RSDs of 1.13%-5.37%. Through the determination of 16 batches of samples, the average content of the above 10 ingredients was in turn (5.724 ± 0.017), (0.273 ± 0.003), (0.854 ± 0.005), (1.228 ± 0.004), (0.496 ± 0.003), (1.287 ± 0.004), (0.137 ± 0.004), (3.624 ± 0.014), (7.366 ± 0.032) and (1.754 ± 0.005) mg/g, respectively. Conclusion The established UPLC method is simple, specific, sensitive, stable, precise, accurate, and reproducible, which can be used for quality control and evaluation of YG.

17.
Article in Chinese | WPRIM | ID: wpr-846016

ABSTRACT

Objective: To establish HPLC-ELSD fingerprint of Zhenwu Decoction(ZWD), screen out the signature components of ZWD through chemical pattern recognition, so as to establish the content determination method of ZWD based on this index. Methods: The fingerprint of 16 batches of ZWD was established by HPLC-ELSD method. The similarity evaluation system of traditional Chinese medicine chromatographic fingerprint (2012 Version) was used for similarity evaluation to determine the common peaks and its attribution. Cluster analysis (CA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to select the index components of ZWD. Results: The fingerprint of ZWD was established, 38 common peaks were confirmed, and the similarity was > 0.95. The results of CA, PCA and OPLS-DA were consistent and the samples were divided into three categories. Benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide II, atractylenolide III, oxypaeoniflorin, albiflorin, paeoniflorin and benzoylpaeoniflorin were identified as the 11 index components with significant difference contribution in different batches of ZWD samples. 6-Gingerol and 6-shogaol were the main active components of ginger, so the above 13 components were taken as the index components of ZWD. The chromatographic peak separation degree and linear relationship were good. The average recovery rate was 96.46%-99.80%, RSD ≤ 3.15%. The mass fraction range of benzoylmesaconine, benzoylaconitine, benzoylhypacoitine, polyporenic acid C, pachymic acid, atractylenolide II, atractylenolide III, oxypaeoniflorin, albiflorin, paeoniflorin, benzoylpaeoniflorin, 6-gingerol, 6-shogaol in 16 batches were 283.93-576.86, 25.05-147.39, 62.96-303.37, 31.24-131.27, 9.76-44.04, 32.15-83.55, 76.55-333.13, 17.48-146.61, 456.58-1554.14, 3 322.48-5 590.01, 158.21-556.50, 525.85-582.92 and 68.52-74.73 mg/g, respectively. Conclusion: The fingerprint combined with PCA, CA and OPLS-DA can comprehensively evaluate the quality of ZWD. This method is stable and reliable, providing reference for the quality evaluation.

18.
Article in Chinese | WPRIM | ID: wpr-807900

ABSTRACT

@#An HPLC-DAD wavelength switching method(240 nm, 280 nm, 316 nm, 403 nm)was developed for simultaneous determination of seven index components: hydroxysafflor yellow A, paeoniflorin, ferulic acid, salvianolic acid B, kaempferol, formononetin and tanshinone IIA in Naoxintong capsule. The qualities of different batches of Naoxintong capsules were evaluated by statistical analysis. Seven index components in 20 batches of Naoxintong capsules were simultaneously determined by HPLC wavelength switching method with Capcell PAK C18 MG II column(250 mm × 4. 6 mm, 5. 0 μm). The mobile phase consisted of methanol-acetonitrile(25 ∶75, A)-0. 1% formic acid aqueous solution(B)with a gradient elution program and a flow rate of 1. 0 mL/min, and the column temperature was 30 °C. The results were analyzed by statistical analysis to evaluate the differences in the quality of Naoxintong capsules. Results showed that the seven active components were well separated and showed good linearity hydroxysafflor yellow A(403 nm)2. 30- 11. 50 mg/L(r=0. 999 2), paeoniflorin(240 nm)8. 81- 44. 05 mg/L(r=0. 999 6), ferulic acid(316 nm)1. 22- 6. 10 mg/L(r=0. 999 6), salvianolic acid B(280 nm)11. 61- 58. 05 mg/L(r=0. 999 4), kaempferol(403 nm)1. 16-5. 80 mg/L(r=0. 999 4), formononetin(240 nm)0. 12- 0. 60 mg/L(r=0. 999 5)and tanshinone IIA(280 nm)2. 28- 11. 40 mg/L(r=0. 999 5). The precision was good and RSD was less than 2. 0%, The repeatability was good and RSD was less than 2. 0%. The stability was good in 24 h. The average recoveries were between 97. 35%- 101. 02% and RSD was less than 2. 0%. The contents of target components in Naoxintong capsules, hydroxysafflor yellow A was 0. 213- 0. 369 mg/g, paeoniflorin was 1. 535- 3. 217 mg/g, ferulic acid was 0. 153- 0. 236 mg/g, salvianolic acid B was 2. 563- 3. 271 mg/g, kaempferol was 0. 103- 0. 181 mg/g, formononetin was 0. 022- 0. 028 mg/g, and tanshinone IIA was 0. 466- 0. 698 mg/g. HPLC wavelength change and gradient elution method was established for simultaneous determination of seven index components in Naoxintong capsule. The method is accurate, sensitive, reliable, and repeatable, and can be used for the quality control of Naoxintong capsule.

19.
Article in Chinese | WPRIM | ID: wpr-804553

ABSTRACT

@#In order to explore the effect and its mechanism of paeoniflorin on PD-L1, a PD-L1 high expression cell model was established in interferon gamma(IFN-γ)-induced HepG2 cells. The cytotoxicity of paeoniflorin was detected by MTT assay. Flow cytometry, ELISA and RT-PCR were performed to detect protein and mRNA levels of PD-L1 regulated by paeoniflorin. In HepG2 cells and Jurkat T cell co-culture system, the expression of IL-2 was detected by ELISA. Besides, T cell proliferation was evaluated by CCK-8 method, and the protein expression levels of PD-L1, JAK and STAT3 after drug treatment were determined by Western blot. These results indicated that paeoniflorin could significantly down-regulate the levels of PD-L1 protein and mRNA. In addition, it increased the number of T cells and the concentration of IL-2 in the co-culture system. Furthermore, paeoniflorin could significantly inhibit the protein expression of JAK and STAT3. Au the above experimental data indicated that paeoniflorin could down-regulate the expression of PD-L1, and its mechanism might be related to the JAK/STAT3 pathway.

20.
Article in Chinese | WPRIM | ID: wpr-802300

ABSTRACT

Objective:To establish the HPLC fingerprint test method for the medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang for investigating the quality transmitting of substance group in preparation process of the medicinal materials-decoction pieces-substance benchmarks, then to evaluate the scientificity and rationality of preparation process of substance benchmarks of Shaoyao Gancaotang by combining with yields of dry extract, transfer rates of effective components and other indexes. Method:Substance benchmarks of Shaoyao Gancaotang was prepared according to the method recorded in ancient medical books, fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks were detected by HPLC, and the contents of effective ingredients were determined. At the same time, the correlation analysis of quality transmitting of substance group during the preparation of substance benchmarks was carried out by combining the yields of dry extract and transfer rates of effective components. Result:The established HPLC fingerprint method has good precision, repeatability and stability, it can be used for the simultaneous determination of fingerprint of medicinal materials, decoction pieces and substance benchmarks. In the fingerprint of substance benchmarks, 16 common peaks were determined by taking liquiritin as the reference peak, of which 6 chromatographic peaks belong to Paeoniae Radix Alba and 11 chromatographic peaks belong to Glycyrrhizae Radix et Rhizoma, 7 major chromatographic peaks were identified. The similarities of fingerprints of 15 batches of medicinal materials, decoction pieces and substance benchmarks of Shaoyao Gancaotang were good by comparing with their respective reference fingerprints(≥ 0.90), the average dry extract rate of 15 batches of substance benchmarks was 24.81%, and no discrete data were found; the average transfer rates of paeoniflorin, liquiritin and glycyrrhizic acid from decoction pieces to substance benchmarks were 79.68%, 63.70% and 51.20%, respectively, and no discrete data were found. Conclusion:In this paper, a scientific and reasonable method for evaluating the process of substance benchmarks of Shaoyao Gancaotang is established by means of the fingerprint method controlled by the whole substance group, the research idea of quality transmitting of substance group in the preparation process, and the evaluation of technical and economic indicators. It can be used as a reference for the evaluation and research of material benchmarks in other famous classical formulas.

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