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1.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 37-44, 2023.
Article in Chinese | WPRIM | ID: wpr-973743

ABSTRACT

ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) identification method of Kaixinsan(KXS) samples, in order to clarify the key quality attributes and provide reference for the quality evaluation of KXS. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of KXS was developed with YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-15 min, 2%-20%A; 15-25 min, 20%-25%A; 25-30 min, 25%-30%A; 30-45 min, 30%-31%A; 45-50 min, 31%-44%A; 50-65 min, 44%-45%A; 65-73 min, 45%-75%A; 73-95 min, 75%-100%A; 95-105 min, 100%A; 105-105.1 min, 100%-2%A; 105.1-120 min, 2%A), the detection wavelength was 320 nm. Ultra high performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) was used to identify the chemical components of KXS with electrospray ionization(ESI), negative ion mode and scanning range of m/z 50-2 000. TLC identification methods for Poria and Ginseng Radix et Rhizoma in KXS were established. ResultThere were 11 common peaks in the specific chromatogram of KXS, attributed to Polygalae Radix, Poria and Acori Tatarinowii Rhizoma. Taking peak 9(α-asarone) as the reference peak, the relative standard deviations of the retention times of 15 batches of KXS samples were<0.2%. A total of 34 compounds were identified by UHPLC-LTQ-Orbitrap MS, including terpenoids, phenylpropanoids, oligosaccharides and ketones. The established TLC had good separation and was rapid, reliable, simple, feasible, suitable for the identification of Poria and Ginseng Radix et Rhizoma in KXS. ConclusionThe specific chromatogram and TLC of KXS are stable and reproducible. The material basis of KXS is basically clarified by MS, which can provide a reference for the development and quality control of KXS.

2.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 34-42, 2023.
Article in Chinese | WPRIM | ID: wpr-972283

ABSTRACT

ObjectiveTo establish the specific chromatogram and thin layer chromatography(TLC) of Qingxin Lianziyin(QXLZY) benchmark samples, in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography(HPLC) specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C18 column(4.6 mm×250 mm, 5 μm) with the mobile phase of acetonitrile(A)-0.2% formic acid aqueous solution(B) for gradient elution(0-10 min, 5%-20%A; 10-20 min, 20%A; 20-25 min, 20%-24%A; 25-40 min, 24%-30%A; 40-55 min, 30%-50%A; 55-65 min, 50%-100%A; 65-75 min, 100%A; 75-75.1 min, 100%-5%A; 75.1-90 min, 5%A), and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry(UHPLC-LTQ-Orbitrap MS) with electrospray ionization(ESI) was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information, the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma, Plantaginis Semen and Scutellariae Radix, and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8(baicalin) as the reference peak. A total of 100 compounds, including flavonoids, organic acids, saponins, amino acids and others, were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix, Lycii Cortex, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple, stable and reproducible, which can provide a reference for the development and quality control of QXLZY.

3.
China Journal of Chinese Materia Medica ; (24): 1280-1288, 2023.
Article in Chinese | WPRIM | ID: wpr-970599

ABSTRACT

This paper compared the differences between two kinds of Bufonis Venenum produced by Bufo gargarizans gargarizans and B. gararizans andrewsi, and verified the rationality of the market value orientation of Bufonis Venenum based on the zebrafish mo-del. Twenty batches of Bufonis Venenum from Jiangsu province, Hebei province, Liaoning province, Jilin province, and Liangshan, Sichuan province, including B. gargarizans gargarizans and B. gararizans andrewsi, were collected. The UHPLC-LTQ-Orbitrap-MS combined with principal component analysis was used to compare the differences between two kinds of Bufonis Venenum. According to the limiting conditions of VIP>1, FC<0.5 or FC>2.0, and peak total area ratio>1%, 9 differential markers were determined, which were cinobufagin, cinobufotalin, arenobufagin, resibufogenin, scillaredin A, resibufagin, 3-(N-suberoylargininyl)-arenobufagin, 3-(N-suberoylargininyl)-marinobufagin, and 3-(N-suberoylargininyl)-resibufogenin. The content of 20 batches of Bufonis Venenum was determined according to the Chinese Pharmacopoeia(2020 edition) by high-performance liquid chromatography, and the 2 batches of Bufonis Venenum, CS7(8.99% of total content) and CS9(5.03% of total content), with the largest difference in the total content of the three quality control indexes of the Chinese Pharmacopoeia(bufalin, cinobufagin, and resibufogenin) were selected to evaluate their anti-liver tumor activity based on the zebrafish model. The tumor inhibition rates of the 2 batches were 38.06% and 45.29%, respectively, proving that only using the quality control indexes of the Chinese Pharmacopoeia as the value orientation of Bufonis Venenum market circulation was unreasonable. This research provides data support for the effective utilization of Bufonis Venenum resources and the establishment of a rational quality evaluation system of Bufonis Venenum.


Subject(s)
Animals , Zebrafish , Bufanolides/analysis , Bufonidae , Chromatography, High Pressure Liquid , Quality Control , Cell Line, Tumor
4.
China Journal of Chinese Materia Medica ; (24): 1124-1131, 2023.
Article in Chinese | WPRIM | ID: wpr-970584

ABSTRACT

A randomized, double-blind, placebo-controlled, multi-center phase Ⅱ clinical trial design was used in this study to recruit subjects who were in line with the syndrome of excess heat and fire toxin, and were diagnosed as recurrent oral ulcers, gingivitis, and acute pharyngitis. A total of 240 cases were included and randomly divided into a placebo group and a Huanglian Jiedu Pills group. The clinical efficacy of Huanglian Jiedu Pills in treating the syndrome of excess heat and fire toxin was evaluated by using the traditional Chinese medicine(TCM) syndrome scale. Enzyme-linked immunosorbent assay(ELISA) was used to determine and evaluate the levels of adenosine triphosphate(ATP), 4-hydroxynonenal(4-HNE), and adrenocorticotropic hormone(ACTH) in plasma of the two groups before and after administration and to predict their application value as clinical biomarkers. The results showed that the disappearance rate of main symptoms in the Huanglian Jiedu Pills group was 69.17%, and that in the placebo group was 50.83%. The comparison between the Huanglian Jiedu Pills group and the placebo group showed that 4-HNE before and after administration was statistically significant(P<0.05). The content of 4-HNE in the Huanglian Jiedu Pills group decreased significantly after administration(P<0.05), but that in the placebo group had no statistical significance and showed an upward trend. After administration, the content of ATP in both Huanglian Jiedu Pills group and placebo group decreased significantly(P<0.05), indicating that the energy metabolism disorder was significantly improved after administration of Huanglian Jiedu Pills and the body's self-healing ability also alleviated the increase in ATP level caused by the syndrome of excess heat and fire toxin to a certain extent. ACTH in both Huanglian Jiedu Pills group and placebo group decreased significantly after administration(P<0.05). It is concluded that Huanglian Jiedu Pills has a significant clinical effect, and can significantly improve the abnormal levels of ATP and 4-HNE in plasma caused by the syndrome of excess heat and fire toxin, which are speculated to be the effective clinical biomarkers for Huanglian Jiedu Pills to treat the syndrome of excess heat and fire toxin.


Subject(s)
Humans , Adrenocorticotropic Hormone , Hot Temperature , Medicine, Chinese Traditional , Adenosine Triphosphate
5.
China Journal of Chinese Materia Medica ; (24): 811-822, 2023.
Article in Chinese | WPRIM | ID: wpr-970551

ABSTRACT

Children's fever is often accompanied by food accumulation. Traditional Chinese medicine believes that removing food stagnation while clearing heat of children can effectively avoid heat damage. To systematically evaluate the efficacy of Xiaoer Chiqiao Qingre Granules(XRCQ) in clearing heat and removing food accumulation and explore its potential mechanism, this study combined suckling SD rats fed with high-sugar and high-fat diet with injection of carrageenan to induce rat model of fever and food accumulation. This study provided references for the study on the pharmacodynamics and mechanism of XRCQ. The results showed that XRCQ effectively reduced the rectal temperature of suckling rats, improved the inflammatory environment such as the content of interleukin-1β(IL-1β), interleukin-2(IL-2), interferon-γ(IFN-γ), white blood cells, and monocytes. XRCQ also effectively repaired intestinal injury and enhanced intestinal propulsion function. According to the confirmation of its efficacy of clearing heat, the thermolytic mechanism of XRCQ was further explored by non-targeted and targeted metabolomics methods based on LTQ-Orbitrap MS/MS and UPLC-QQQ-MS/MS. Non-target metabolomics analysis of brain tissue samples was performed by QI software combined with SIMCA-P software, and 22 endogenous metabolites that could be significantly regulated were screened out. MetaboAnalyst pathway enrichment results showed that the intervention mechanism was mainly focused on tyrosine metabolism, tricarboxylic acid cycle, inositol phosphate metabolism, and other pathways. At the same time, the results of targeted metabolomics of brain tissue samples showed that XRCQ changed the vitality of digestive system, and inhibited abnormal energy metabolism and inflammatory response, playing a role in clearing heat and removing food stagnation from multiple levels.


Subject(s)
Animals , Rats , Rats, Sprague-Dawley , Hot Temperature , Tandem Mass Spectrometry , Metabolomics , Food , Fever , Interferon-gamma
6.
China Journal of Chinese Materia Medica ; (24): 3074-3085, 2023.
Article in Chinese | WPRIM | ID: wpr-981438

ABSTRACT

The tissue distribution of Qingfei Paidu Decoction was studied by HPLC-MS/MS in vivo. Hypersil GOLD C_(18) column(2.1 mm×50 mm, 1.9 μm) was used for gradient elution with acetonitrile as the mobile phase A and 0.1% formic acid solution as the mobile phase B. High-resolution liquid chromatography-mass spectrometry in both positive and negative ion scanning mode and multiple response monitoring(MRM) mode was employed to analyze the behaviors of the active components of Qingfei Paidu Decoction in diffe-rent tissues. The results showed that 19, 9, 17, 14, 22, 19, 24, and 2 compounds were detected in plasma, heart, liver, spleen, lung, kidney, large intestine, and brain, respectively. The compounds belonged to 8 groups, covering 14 herbs in the prescription. After administration with Qingfei Paidu Decoction, the compounds were rapidly distributed in various tissues, especially in the lung, liver, large intestine, and kidney. The majority of the compounds displayed secondary distribution. This study comprehensively analyzed the distribution rules of the main active components in Qingfei Paidu Decoction and provided a basis for the clinical application.


Subject(s)
Chromatography, High Pressure Liquid , Tandem Mass Spectrometry , Tissue Distribution , Drugs, Chinese Herbal
7.
China Journal of Chinese Materia Medica ; (24): 1995-2007, 2022.
Article in Chinese | WPRIM | ID: wpr-928138

ABSTRACT

Quality evaluation of Chinese medicinal decoction pieces is vital for the development of the downstream industries, and is an important channel for implementing the strategy of "higher quality, higher price, and priority for the high quality" for traditional Chinese medicine. At the moment, the quality of Chinese medicinal decoction pieces is mainly evaluated based on chemical component examination. Considering the weak preliminary research foundation and poor research conditions, traditional experience-based evaluation is undervalued in the quality rating of Chinese medicinal decoction pieces. However, traditional experience is a summary of the quality of Chinese medicinal materials based on clinical experience, which thus can be a potential basis for the quality evaluation of the decoction pieces. It is a challenge in the evaluation of Chinese medicinal decoction pieces to objectify the traditional experience-based evaluation from multiple aspects such as chemistry, effect, and characterization via modern techniques. Therefore, this study developed the "experience-ingredients-activity-electronic sensing" evaluation system for Chinese medicinal decoction pieces on the basis of experience-based assessment, chemical ingredients that can truly reflect the traditional experience, biological effect assessment, and electronic sensory evaluation, which is expected to quantify the traditional experience of quality evaluation of Chinese medicinal decoction pieces via chemistry, biology, and sensory simulation. The evaluation system can serve as a reference for clinical experience-based quality evaluation of Chinese medicinal decoction pieces.


Subject(s)
China , Drugs, Chinese Herbal , Electronics , Medicine, Chinese Traditional , Restraint, Physical
8.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 127-133, 2022.
Article in Chinese | WPRIM | ID: wpr-940838

ABSTRACT

ObjectiveAlmond, which is bitter in taste, contains traces of toxic substances. For the sake of the safety of prescriptions containing this medicinal material, the processing method of "soaking in boiling water" was selected. Moreover, through literature research and network pharmacology, the characteristic index of this medicinal material was determined. On this basis, a method was established for the determination of amygdalin in Qingfei Paidu Granules (QFPD) and the transfer rate of it in the processing of this prescription was monitored, aiming at improving the quality control system of QFPD. MethodThe high performance liquid chromatography conditions are as follows: YMC Triart C18 column (4.6 mm × 150 mm, 5 µm), mobile phase of methanol-water with flow of 1.0 mL·min-1, column temperature of 35 ℃, and detection wavelength of 210 nm. ResultThe linear curve fitted well and the average recovery of amygdalin was 97.74% with RSD of 4.3%. The transfer rates of amygdalin from the medicinal material to the extract, from extract to concentrate, and from concentrate to granules were investigated with this method. The result showed that the average transfer rate from the medicinal material to the granules was (60±3.91)%. The comparison of transfer rate between the processes suggesting that the extraction of the medicinal material might be the key part influencing the prescription preparation. ConclusionThe method is simple, sensitive, reproducible, stable, and accurate, and the index is reasonable. Thus, the method can be used for the quality control of QFPD and determination of transfer rate of components in the preparation of QFPD. This study further improves the quality control standard of almond in QFPD, which can serve as a reference for the clinical application of QFPD.

9.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 224-232, 2021.
Article in Chinese | WPRIM | ID: wpr-906475

ABSTRACT

Qingxin Lianzi Yin (QXLZY), as an ancient classical formula for clearing the heart and nourishing the Yin, was composed of nine herbs (Scutellariae Radix, Ophiopogonis Radix, Lycii Cortex, Plantaginis Semen, Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle, Nelumbinis Semen, Poria, Astragali Radix and Ginseng Radix et Rhizoma), coming from Prescriptions of the Bureau of Taiping People's Welfare Pharmacy. It could clear away the heart-fire, promote the interaction of the heart and kidney, replenish Qi and Yin, and stop strangury-turbidity. It was used to treat gonorrhea of urination, seminal emission, restlessness, wasting-thirst and so on. At present, the usage and dosage of QXLZY and its addition and subtraction are different in clinical practice. Most of the studies just focus on its clinical efficacy, and there is few review literature reflecting its historical evolution. Based on this, this paper systematically clarified the historical evolution, composition, preparation, interpretation, function, and modern clinical application of QXLZY. This work has been explained the historical evolution of QXLZY, and found that it was wildly used in modern clinical, especially suitable for the treatment of chronic urinary system diseases. At the same time, QXLZY also had significant therapeutic effects on neurasthenia, stomatitis, diabetic nephropathy and other aspects. Through the comprehensive analysis of ancient and modern literature, this work explores the true connotation of QXLZY from the perspective of traditional Chinese medicine theory, which can point out the direction of the clinical application and positioning of this famous classical formula after it comes into the market, and also can provide reference basis for its subsequent in-depth research and development.

10.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 97-103, 2021.
Article in Chinese | WPRIM | ID: wpr-905868

ABSTRACT

Objective:To explore effects of different extracts and monomers of <italic>Lepidium meyenii </italic>(Maca) on the proliferation of mouse splenic lymphocytes and induction of interleukin-2 (IL-2) and tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) by observing their immunomodulatory effects. Method:An octadecylsilyl (ODS) column was used to enrich the methanol extract of <italic>L. meyenii</italic> in stages to obtain six fractions and three monomers. Different groups of extracts and monomers of <italic>L. meyenii </italic>at different doses were set up. Cell counting Kit-8 (CCK-8) was used to detect the effect on the proliferation of mitogen-free, concanavalin A (Con A)-induced, and lipopolysaccharides (LPS)-induced mouse splenic lymphocytes. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of IL-2 and TNF-<italic>α</italic>. Result:<italic>L. meyenii </italic>extracts Fr<sub>3</sub> and Fr<sub>6</sub>, and monomers <italic>N</italic>-benzyl hexadecanamide and 1,2-dihydro-4-carboxaldehyde-3-benzyl-<italic>N</italic>-hydroxypyridine slightly promoted the proliferation of Con A-induced T lymphocytes and LPS-induced B lymphocytes (<italic>P</italic><0.01) as compared with the conditions in the model group. <italic>L. meyenii</italic> extracts and monomers significantly induced the secretion of IL-2 and TNF-<italic>α</italic> by splenic lymphocytes (<italic>P</italic><0.01). Conclusion:<italic>L. meyenii</italic> extracts and monomers can achieve immunological enhancement by promoting the secretion of IL-2 and TNF-<italic>α</italic>, and facilitate the proliferation of splenic lymphocytes. The active components are presumedly macamides and pyridine alkaloids, and the specific mechanism still needs to be further explored.

11.
China Journal of Chinese Materia Medica ; (24): 4774-4781, 2021.
Article in Chinese | WPRIM | ID: wpr-888184

ABSTRACT

Indolealkylamines(IAAs) are the main hydrophilic substances in toad skin, mainly including free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine. In this study, the LPS-activated neutrophils were used to investigate the structure-activity relationship and anti-inflammatory mechanism of the above-mentioned five monomers from the toad skin in vitro. The neutrophils were divided into the control group, model group(1 μg·mL~(-1) LPS), positive drug group(100 μg·mL~(-1) indometacin), as well as the low-(50 μg·mL~(-1)), medium-(100 μg·mL~(-1)) and high-dose(200 μg·mL~(-1)) free N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine, dehydrobufotenine, and binding bufothionine groups. The levels of IL-6, TNF-α and IL-1β in the neutrophil supernatant of each group was measured by enzyme-linked immunosorbent assay(ELISA) after LPS stimulation, followed by the detection of apoptosis in each group after Annexin V/PI staining. The protein expression levels of caspase-3, Bax, Bcl-2, beclin1, LC3-I, and LC3-Ⅱ were assayed by Western blot. The results showed that IAAs reduced the excessive secretion of inflammatory cytokines caused by LPS compared with the model group. Besides, the activity of each free IAAs(N-methyl-5-hydroxytryptamine, bufotenine, bufotenidine and dehydrobufotenine), especially bufotenine, was stronger than that of the binding bufothionine. As revealed by Annexin V/PI staining, LPS delayed the early apoptosis of neutrophils compared with the control group, while bufotenine promoted the apoptosis of neutrophils in a dose-dependent manner, which might be related to the elevated expression of apoptosis-related protein Bax/Bcl-2. In addition, LPS activated the autophagy pathways in neutrophils. This study confirmed the efficacy of IAAs in reducing the secretion of inflammatory cytokines in neutrophils induced by LPS for the first time. For instance, bufotenine exerts the anti-inflammatory effect possibly by inducing the apoptosis of neutrophils.


Subject(s)
Animals , Anti-Inflammatory Agents/pharmacology , Apoptosis , Bufonidae , Lipopolysaccharides/toxicity , Neutrophils , Skin
12.
China Journal of Chinese Materia Medica ; (24): 6163-6177, 2021.
Article in Chinese | WPRIM | ID: wpr-921774

ABSTRACT

This study aimed to qualitatively analyze the chemical components in Xiaoer Chiqiao Qingre Granules(XRCQ) by UHPLC-LTQ-Orbitrap-MS/MS and identify its material basis. The absorbed components in plasma were combined for exploring the potential action mechanism by integrated network pharmacology. ACQUITY UPLC HSS T3(2.1 mm×100 mm, 1.8 μm) column and mobile phase system of 0.1% formic acid solution(A)-acetonitrile(B) were used for gradient elution, followed by high resolution liquid chromatography-mass spectrometry in both positive and negative ion scanning modes. According to the precise relative molecular mass and MS/MS fragment ions, a total of 124 chemical components were identified in XRCQ by the comparison with references and literature reports, among which 29 compounds were completely confirmed by comparison with reference substances. Then, the main absorbed components of XRCQ in plasma were also analyzed and clarified by UHPLC-LTQ-Orbitrap-MS/MS. BATMAN-TCM and SwissTargetPrediction were used for target prediction of absorbed components in plasma. Following the plotting of association network with Cytoscape 3.8.2, the core targets were subjected to GO and KEGG pathway enrichment analysis and a component-target-pathway network was constructed. A total of eight main targets of XRCQ against fever in children were obtained together with eight absorbed components in plasma, including glycyrhydinic acid, hesperidin, emodin, reticuline, daidzein, magnolignan C, magnolignan A, and magnolaldehyde D. It was inferred that XRCQ might improve alimentary system abnormality, inflammatory response, oxidative stress, and endocrine disorder through tumor necrosis factor, PI3 K-AKT, and other signaling pathways. The present study comprehensively expounded the chemical profiles of XRCQ and the main absorbed components in plasma and predicted the potential mechanism of XRCQ based on integrated network pharmacology, which has provided certain theoretical reference for the clinical application of XRCQ.


Subject(s)
Child , Humans , Chromatography, High Pressure Liquid , Chromatography, Liquid , Drugs, Chinese Herbal , Network Pharmacology , Tandem Mass Spectrometry
13.
China Journal of Chinese Materia Medica ; (24): 3035-3044, 2020.
Article in Chinese | WPRIM | ID: wpr-828018

ABSTRACT

UHPLC-LTQ-Orbitrap-MS was developed for the identification of chemical constituents in Qingfei Paidu Decoction, which will clarify its material basis. ACQUITY UHPLC HSS T3 chromatography column(2.1 mm×100 mm, 1.8 μm) was used with 0.1% formic acid(B)-acetonitrile(A) as the mobile phase in gradient elution. The decoction was detected by high-resolution liquid chromatography-mass spectrometry equipped with an ESI ion source in positive and negative mode. Based on the accurate mass measurements, retention time, mass fragmentation patterns combined with comparison of reference and literature reports, a total of 87 major compounds including 43 flavonoids, 9 alkaloids, 4 triterpenoid saponins, 1 sesquiterpene, 2 coumarins, 10 phenolic acids and 18 other compounds were tentatively screened and characterized. UHPLC-LTQ-Orbitrap-MS was employed to comprehensively elucidate the chemical components in Qingfei Paidu Decoction, which basically covered 20 Chinese medicines except gypsum in Qingfei Paidu Decoction. These collective results provide a scientific basis for further research on the quality control standard of Qingfei Paidu Decoction.


Subject(s)
Chromatography, High Pressure Liquid , Coumarins , Drugs, Chinese Herbal , Flavonoids , Mass Spectrometry
14.
China Journal of Chinese Materia Medica ; (24): 158-166, 2019.
Article in Chinese | WPRIM | ID: wpr-771503

ABSTRACT

In order to find the endogenous potential biomarkers of in vitro hepatic injury caused by NCTD-Na and elucidate the mechanism of hepatic injury of NCTD-Na,ultra-high performance liquid chromatography coupled quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used for lipidomics detection.Multivariate statistical analysis was used to study the endogenous lipid metabolic changes of human normal liver cells LO2 injury after the treatment with sodium norcantharidate(NCTD-Na).The results showed that the half maximal inhibitory concentration(IC50) of NCTD-Na was 0.034 mmol·L-1.A total of 280 differential metabolites were found between the control group and the low-dose group,with VIP > 2.0 and P 2.0 and P 2.0,P<0.05,RSD<30% and in a dose-dependent manner.It was found that most of the above differential metabolites were lipid metabolites after the analysis of simple preparnation methods and database search.A total of 32 potential biomarkers were identified,including 3 phosphatidylcholine(PC),5 lysophosphatidylcholine(Lyso PC),3 ceramide(Cer),1 sphingomyelin(SM),1 phosphatidylethanolamine(PE),10 lysophosphatidylethanolamine(LysoPE),4 diacylglycerol(DG),1 Phosphatidic acid(PA),1 lysophosphatidic acid(Lyso PA),1 phosphatidyl glycerol(PG),1 fatty acid hydroxy fatty acid(FAHFA) and 1 phosphatidylserine(PS).The changes of PCs,Cers,SM,PE and DGs were closely related liver protection,DNA methylation and self-repair in hepatocytes,apoptosis,methylation and detoxification of carcinogens,as well as lipid peroxides production process.Also,they had impact on the proliferation of hepatocytes,differentiation and gene transcription disorders.Cells stimulated by NCTD-Na could promote the production of PA as well as the synthesis and catabolism of FAHFA in a variety of ways.The levels of Lyso PCs,LysoPEs and Lyso PA were correlated with PCs,PE and PA;PE and PS might have valgus during apoptosis,triggering phagocytosis.


Subject(s)
Humans , Bridged Bicyclo Compounds, Heterocyclic , Pharmacology , Cells, Cultured , Hepatocytes , Metabolism , Lipid Metabolism , Lipids , Tandem Mass Spectrometry
15.
China Journal of Chinese Materia Medica ; (24): 3006-3011, 2018.
Article in Chinese | WPRIM | ID: wpr-687354

ABSTRACT

To study the pharmacokinetics of active ingredients (alkaloids, iridoids and flavonoids) in Huanglian Jiedu decoction (HLJDD) in Alzheimer's disease (AD) model rats, and investigate its mechanism in treatment of AD. All of rats were divided into normal control group (=6), shame operation group (=6) and model group (=12). Rats in shame operation group received daily subcutaneous injection of D-galactose (D-gal 50 mg·kg⁻¹) for a total of 45 d to induce subacute aging model. Based on the operation of shame operation group, the rats in model group were given with an injection of Aβ₂₅₋₃₅ (4.0 g·L⁻¹)-ibotenic acid (2.0 g·L⁻¹) into the nucleus basalis magnocellularis. Then rats in model group were divided into HLJDD one day administration group (=6) and HLJDD one week group (=6). The plasma concentration of alkaloids, iridoid and flavonoids was determined by liquid chromatography tandem mass spectrometry (LC-QQQ-MS) at different time points. The levels of seven inflammatory factors (MIP-2, IL-1β, IL-2, IL-8, IL-10, IL-13, TNF-α) in cerebrospinalfluid (CSF) were measured by Bio-Plex multi-factor detection technology. Iridoids in AD model rats, with high bioavailability, were easily absorbed and eliminated. The plasma concentration of alkaloid was lower, and the AUC (area under the curve) was higher in HLJDD one week group than that in HLJDD one day group. The plasma concentration-time curves of flavonoids showed obvious bimodal phenomena. After the gastric administration of HLJDD, the inflammatory factors in CSF of AD rats demonstrated a callback trend, including IL-1β/IL-10 (<0.05) with significant difference. The pharmacokinetic behaviors of iridoids, alkaloids and flavonoids (41 compounds) in AD model rats were fundamentally elucidated, and HLJDD can improve the central inflammatory status of AD rats by regulating the levels of inflammatory factors.

16.
Chinese Journal of Nephrology ; (12): 30-35, 2018.
Article in Chinese | WPRIM | ID: wpr-711085

ABSTRACT

Objective To explore the effect of acteoside (one of the ingredients of Total Glycosides Extracted from Rehmannia capsules) on treatment of proteinuria and protection of podocytes.Methods In this study,puromycin nephropathy rat model was successfully established.After detecting the degree of proteinuria,the expression of podocyte injury markers and the degree of podocyte foot process fusion were investigated by electron microscope.In addition,puromycin treated podocyte injury model was also successfully established in vitro.Podocyte viability,migration,cytoskeleton and injury marker were detected.Results In vivo study showed that acteoside could effectively reduce proteinuria (P < 0.05),restore the expression of podocyte injury markers such as nephrin and synaptopodin (all P < 0.05),and alleviate the degree of podocyte foot process fusion.In vitro study showed that acteoside could effectively restored podocyte viability (P < 0.05),reduce abnormal migration ability (P < 0.05),protecte cytoskeleton and restore the expression of podocyte injury marker nephrin (P < 0.05).Conclusions This study confirms that acteoside can reduce the degree of proteinuria in puromycin nephropathy rat model in vivo and alleviate the degree of podocyte injury in vitro as well as enrich the molecular mechanism of Total Glycosides Extracted from Rehmannia capsules in treatment of proteinuria.

17.
Chinese Journal of Dermatology ; (12): 654-657, 2017.
Article in Chinese | WPRIM | ID: wpr-686575

ABSTRACT

Objective To investigate characteristic skin lesions and typical histopathological changes of adult-onset Still's disease(AOSD) for its early diagnosis and treatment.Methods Clinical data were collected from 8 patients with AOSD,and analyzed retrospectively.Results All the patients had transient rashes and persistent papules/plaques during the course of disease.Of the 8 patients,1 had urticaria-like rashes,3 had dermatomyositis-like rashes,and 1 had prurigo pigmentosa-like rashes.Biopsies were carried out at the sites where transient rashes or persistent papules/plaques occurred.Histopathological findings showed necrotic keratinocytes in the upper prickle cell layer,and perivascular infiltration of neutrophils and lymphocytes in the upper dermis.Conclusion The skin lesions and histopathological changes of AOSD are characteristic,which can provide clues to the early diagnosis of AOSD.

18.
China Journal of Chinese Materia Medica ; (24): 3767-3772, 2016.
Article in Chinese | WPRIM | ID: wpr-307088

ABSTRACT

Fifteen compounds were isolated from the toad skin by a combination of various chromatographic methods including macroporous resin, silica gel, ODS and semi-preparative HPLC. Their structures were identified as 4,5-dimethyl-1,3,4,5-tetrahydropyrrolo[4,3,2-de]quinolin-6-ol(1), serotonin(2), N-methyl serotonin(3), O-methyl bufotenine(4), 1,2,3,4-tetrahydro-6-hydroxy-β-carboline(5), O-methylserotonin(6), glycinebetaine(7), caffeine(8), bufotenine(9), shepherdine(10), tryptophan(11), (5-hydroxy-1H-indol-3-yl)acetic acid(12), 5-hydroxy tryptophol(13), 2-methyl-6-hydroxy-1,2,3,4-tetrahydro-β-carboline(14), bufothionine(15). Among them, compound 1 was a new compound,compound 5 was a new natural product. Compounds 4-8 and 10-14 were separated from toad skin for the first time.

19.
China Journal of Chinese Materia Medica ; (24): 2669-2674, 2016.
Article in Chinese | WPRIM | ID: wpr-275187

ABSTRACT

In this study, 10 samples of parasites, cursive, and the whole from six different species of Cordyceps were determined and compared by HPLC and LC-MS methods. Uridine, adenosine, and cordycepin were selected as the main evaluation index. The anti-fibrotic activity of different species Cordyceps extracts was observed using in vitro TGF-β1-induced ECM accumulation in human embryonic fibroblasts CCC-ESF-1. The results demonstrated that the number of atoms and hyphae ingredients of different species showed little difference, however, the content distribution of each component has obvious significance. The in vitro anti-fibrotic activities of different species were as follow: Cordyceps flower > Cicada Cordyceps (Cicada flower)> Silkworm Cordyceps> Tussah Cordyceps>natural Cordyceps. Our preliminary data could serve as reference for the discovery of artificial alternatives of natural Cordyceps.

20.
Journal of Forensic Medicine ; (6): 135-139, 2015.
Article in Chinese | WPRIM | ID: wpr-983981

ABSTRACT

The event-related potential (ERP) is considered as one of the most effective methods to study and analyze objectively human mental activity based on nerve electrophysiology. At present, ERP is not only used in the study of lie detection, but also in the clinical medicine for the cognitive assessment on patients with cerebrovascular disease, dementia or traumatic brain injury and auxiliary diagnosis of mental illness. With the further development of ERP detection technology, it would have a wider application prospect in the field of forensic medicine.


Subject(s)
Humans , Evoked Potentials/physiology , Forensic Medicine/trends
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