Study on improvement mechanism of caudatin on liver injury in rats / 中国药房

OBJECTIVE To investigate the improvement mechanism of caudatin on liver injury of rats. METHODS SD rats were randomly divided into blank group， model group， caudatin low-dose and high-dose groups （25， 50 mg/kg）， with 6 rats in each group. Diethylnitrosamine （DEN） was injected intraperitoneally three times per week for eight weeks to establish liver injury model of rats. At 5th week of modeling， the rats received relevant medicine or 0.5% sodium carboxymethylcellulose intragastrically for 4 weeks. The levels of liver function indexes [alanine transaminase （ALT）， aspartate transaminase （AST）， total protein （TP） and total bilirubin （TBI）] and inflammatory factors [interleukin （IL-6）， tumor necrosis factor α （TNF-α）， IL-1β] in serum were detected； the histopathological morphological changes of rat liver were observed； the positive protein expressions of nuclear factor κB （NF-κB） and 78 kDa glucose regulatory protein （Grp78） in liver tissue were also determined； the expressions of endoplasmic reticulum stress-related protein Grp78， C/EBP homologous protein （CHOP）， activating transcription factor 6 （ATF6） and inositol requiring enzyme 1α （IRE1α） and the level of protein kinase R-like endoplasmic reticulum kinase robertluoyi@126.com （PERK） in liver tissue were detected. RESULTS Compared with blank group， serum levels of ALT， AST， TBI， IL-6， TNF-α and IL-1β and positive expressions of NF-κB and Grp78 in liver tissue as well as protein expressions of Grp78， CHOP， ATF6 and IRE1α， PERK protein phosphorylation level were all increased significantly in model group （P＜0.05）， while the serum level of TP was decreased significantly （P＜0.05）. The disordered structure of liver lobule， swollen liver cells， unclear intercellular boundary were observed and accompanied by inflammatory cell infiltration. Compared with model group， most of the above indexes were significantly reversed in caudatin groups （P＜0.05）； the structure of hepatic lobule was relatively complete and clear， the cells were arranged orderly， and the infiltration of inflammatory cells was also reduced. CONCLUSIONS Caudatin has a significant improvement effect against DEN-induced liver injury in rats， the mechanism of which may be associated with inhibiting endoplasmic reticulum stress and inflammatory reaction.

[Product improvement and secondary development of Pudilan Xiaoyan Oral Liquid based on clinical medication].

Pudilan Xiaoyan Oral Liquid is widely used in clinical applications, with safe and effective results. Its coverage rate in the national first, second and third grade hospitals is as high as 71%. In this study, we analyzed and summarized the research progress on the material basis, quality control, production process and clinical medication of Pudilan Xiaoyan Oral Liquid based on the clinical diseases(parotitis, tonsillitis, pharyngitis), and deeply explored the intrinsic quality improvement and secondary development of Pudilan product. Pharmacodynamic material basis of Pudilan Xiaoyan Oral Liquid was explored through the network pharmacology technology and quality control indicators of the production process were optimized by cell anti-inflammatory experiments. Through these techno-logies, it would be more specific, scientific and effective to carry out process optimization of each link and multidimensional quality control of the whole process. The dosage and oral compliance for special patients(children) were explored, providing a reference for clinical pediatric medication of Pudilan Xiaoyan Oral Liquid. Simultaneously, it is helpful to expand the application market by developing Pudilan daily chemical products, and promote the traditional Chinese medicine products in terms of curative effect and daily life.

[Investigation of active components and mechanism of Pudilan Xiaoyan Oral Liquid based on network pharmacology].

To investigate the active components/ingredients of Pudilan Xiaoyan Oral Liquid based on the network pharmacology technology, and analyze the network data of its potential targets and mechanisms. The active ingredient screening, protein interaction analysis and pathway annotation were used to further optimize its active components and potential targets, and clarify the pharmacodynamic substance basis and mechanism of Pudilan Xiaoyan Oral Liquid. Through this technique, we screened out 41 active ingredients in Pudilan Xiaoyan Oral Liquid, mainly including 16 alkaloid components, 13 organic acid components, 11 flavonoid components and 1 coumarin component such as chicoric acid, chlorogenic acid, oroxindin, rutin, corynoline, and esculetin. In addition, 6 targets for parotitis, 48 targets for tonsillitis, and 22 targets for pharyngitis were screened. A total of 22 disease signaling pathways are involved, including 4 pathways closely related to inflammation. The IL-17 signaling pathway had the highest D(degree) value and may be most closely related to inflammatory diseases. Through network data excavating, we initially explored the main active components/ingredients of Pudilan Xiaoyan Oral Liquid, clarified the pharmacodynamic basis of Pudilan Xiaoyan Oral Liquid treatment-related diseases and its key mechanism of action in this study, hoping to provide a theoretical basis for clinical research, and at the same time, lay the foundation for deep research and promotion of Pudilan Xiaoyan Oral Liquid product.

[Identification of quality markers for Pudilan Xiaoyan Oral Liquid].

On the basis of anti-inflammatory activity, combined with multiple indicators, the quality markers of Pudilan Xiaoyan Oral Liquid were screened and determined for quality control. Lipopolysaccharide(LPS) was used to induce normal human bronchial epithelial cell(NHBE) inflammation model. The anti-inflammatory effects of the main chemical components in Pudilan Xiaoyan Oral Liquid were examined one by one, and the pharmacodynamic basis for the overall anti-inflammatory efficacy of Pudilan Xiaoyan Oral Liquid was clarified to identify the quality markers of Pudilan Xiaoyan Oral Liquid and the contents of the quality markers were determined by high performance liquid chromatography(HPLC). The results showed that adenosine, epigoitrin, chlorogenic acid, caffeic acid, cichoric acid, corynoline, baicalin, wogonoside, wogonin and oroxylin A had a certain regulatory effect on inflammatory factor tumor necrosis factor(TNF-α), interleukin(IL-1ß) and IL-6 at specific concentrations in a dose-dependent manner. Considering the factors such as the IC_(50) value of each monomer component and the comprehensiveness of the quality control components, we proposed to use adenosine, cichoric acid, corynoline, baicalin and wogonin as quality markers of Pudilan Xiaoyan Oral Liquid. The contents of the five components were determined by HPLC, and the results showed that they were relatively stable in three batches of Pudilan Xiaoyan Oral Liquid. In this study, the quality control components selected by the anti-inflammatory activity test have a clear material basis, covering all four active pharmaceutical ingredients, which can fully reflect the quality of Pudilan Xiaoyan Oral Liquid, and effectively improve the quality control standard of Pudilan Xiaoyan Oral Liquid.

[Improvement of compliance of Pudilan Xiaoyan Oral Liquid based on comprehensive analysis of bad taste of traditional Chinese medicine oral liquid preparation].

From "good efficacy with a bad taste" to "good efficacy with no bitterness" and then to "good efficacy with a good taste" is the only way to develop oral liquid preparations of traditional Chinese medicine, and "good medicine is beneficial to disease" is the only way for the development of oral liquid preparations of traditional Chinese medicine. Based on the analysis of the causes, the sources of bitterness, the formation principles and their solutions of traditional Chinese medicine oral liquid preparations, we explored the causes of the bad taste and the material basis of bitterness of Pudilan Xiaoyan Oral Liquid, and applied the solutions in improving the taste of Pudilan products. The overall taste of Pudilan Xiaoyan Oral Liquid was improved by modifying the original product taste, enhancing the process and changing the dosage form, which improves the compliance of the patients who take the medicine, and better serve the clinical medication.

[Source control of Pudilan Xiaoyan Oral Liquid--research on quality of Taraxaci Herba based on ultra high performance liquid chromatography fingerprint].

Pudilan Xiaoyan Oral Liquid has been widely used in the clinical treatment of inflammatory diseases such as upper respiratory tract infections. Taraxaci Herba, as the monarch medicine in Pudilan Xiaoyan Oral Liquid, due to its multi-source, multi-origin characteristics, and the difference in the content of active ingredients in different medicinal parts, has become a potential factor for the unstable quality among different batches of Pudilan Xiaoyan Oral Liquid. In this paper, Thermo Scientific Vanquish ultra-high-performance liquid chromatography(UPLC) system was used, and the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition) issued by National Pharmacopoeia Commission was used for processing and analysis. The main common peaks were identified and contents were determined by comparison with reference substances. Fingerprints of Taraxaci Herba medicinal materials from different origins were established. 13 common peaks were identified, and 29 batches of samples from five origins had similarities above 0.90. At the same time, an ultra-high performance liquid chromatography method was developed for the determination of monocaffeoyl tartaric acid, chlorogenic acid, caffeic acid, chicoic acid, and luteolin in Taraxaci Herba. The quantitative analysis conditions were verified by methodology, and the average sample recovery was 97.30%-101.8%. The results showed that the content of the same ingredient in Taraxaci Herba from different origins and different medicinal parts was obviously different, and the fluctua-tion range was also different for different ingredients. The establishment of UPLC fingerprints for Taraxaci Herba from different regions combined with multi-component content determination methods provides a reference for improving the quality control of Taraxaci Herba medicinal materials, and also provides a source guarantee for the quality improvement of Pudilan Xiaoyan Oral Liquid.

Effect of Recipes for Tonifying Kidney and Replenishing Qi on Learning and Memory-related Genes in Aged Rats / 中国实验方剂学杂志

Objective:To observe the effects of recipes for tonifying kidney and replenishing Qi, Zuoguiwan (ZG) and Yiqi Congming Tang(YQ) on memory capacity, expressions of learning and memory-related genes expression, and explore the changes in relevant epigenetic modification enzymes. Method:SD male rats with natural aging (24 months old) were used as animal models and randomly divided into aged control group, aged ZG group(12.12 g·kg-1), aged YQ group(10.18 g·kg-1), aged compound group(11.15 g·kg-1) and aged antagonist RU38486 group(5×10-3g·kg-1). Another 5 months old male SD rats were included as the young control group. Morris water maze method was used to observe the spatial learning and memory ability of the rats. The co-localizations of histone deacetylase 2 (HDAC2) and methyl CpG binding protein 2 (MeCP2) in hippocampus of rats in each group were observed by laser confocal microscope. The changes in expressions of glucocorticoid receptor (GR), synapsin1(Syn-1), HDAC2, and histone acetyltransferase 1(HAT1) proteins in hippocampus of each group were detected by Western blot, and mRNA expression of HDAC2 was detected by Real-time fluorescence quantitative polymerase Chain reaction (Real-time PCR). At the same time, the effects of ZG, YQ and compound decoction in alleviating the above-mentioned abnormal changes were observed. Result:Compared with the young control group(control group), the latency of the aged control group was significantly prolonged (PPPPPPPConclusion:ZG group, YQ group, and compound group can improve the spatial learning and memory abilities of aged rats by increasing the expression of learning-memory-associated protein GR and epigenetic modification enzyme HAT1, and reducing the expression of HDAC2 and the co-localization of HDAC2 protein and MeCP2 in the nucleus.

Characterization of sediment of water extract of Guizhi decoction and its effect on relevant compound (fractions) in decoction / 中国中药杂志

In order to study the effect of sediment of water extract of Guizhi decoction on the stability, clarity and peaks area, and characterize the chemical composition of the sediments, HPLC and MS methods were established. Through comparison of the common peak areas and the turbidity value of water extract and filtrate, the sediments could greatly change the common peak areas of the decoction (for more than 5 times of the study standard); at the same time, the turbidity value of the decoction could increase by （38.66±1.57）% in 48 h [particularly by （24.54±1.68）% in 6 h]. The test indicated that the sediments had an effect on the stability and clarity under the test conditions in Guizhi decoction. The study confirmed that the sediments were mainly derived from Cassia twig, Paeonia lactiflora and Glycyrrhiza uralensis. On the basis of the reference information, the accurate molecular weight and fragment ion information provided by LC-MS were analyzed, the molecular formula of sediments components A-F were determined, and the possible structural information of components B, C, D and F were deduced. It was suggested that the multi-index, multi-target and multi-angle analysis could ensure the quality of traditional Chinese medicine and the effect of clinical medication. The study also suggested the effect of the sediments on clinical application and the preparation of traditional Chinese medicine.

Ideas and methods on efficient screening of traditional medicines for anti-osteoporosis activity based on M-Act/Tox integrated evaluation using zebrafish / 中国中药杂志

The increasingly apparent liver injury problems of bone strengthening Chinese medicines have brought challenges for clinical application, and it is necessary to consider both effectiveness and safety in screening anti-osteoporosis Chinese medicines. Metabolic transformation is closely related to drug efficacy and toxicity, so it is significant to comprehensively consider metabolism-action/toxicity(M-Act/Tox) for screening anti-osteoporosis Chinese medicines. The current evaluation models and the number of compounds(including metabolites) severely restrict efficient screening in vivo. By referring to previous relevant research and domestic and abroad literature, zebrafish M-Act/Tox integrative method was put forward for efficiently screening anti-osteoporosis herb medicines, which has organically integrated zebrafish metabolism model, osteoporosis model and toxicity evaluation method. This method can break through the bottleneck and blind spots that trace compositions can't achieve efficient and integrated in vivo evaluation, and realize both efficient and comprehensive screening on anti-osteoporosis traditional medicines based on in vivo process taking both safety and effectiveness into account, which is significant to accelerate discovery of effective and safe innovative traditional Chinese medicines for osteoporosis.

Synergistic effect of Epimedii Folium fried with suet oil for warming kidney and enhancing yang in dosage form of self-assembled micelles / 中国中药杂志

The fried method with suet oil, which can strengthen the effect of Epimedii Folium in warming kidney and enhancing yang, has been widely used in the processing of Epimedii Folium in traditional Chinese medicine. In this paper, we adopted hydrocortisone-induced kidney-yang deficiency rat models, and took physical signs, serum testosterone, corticosterone levels and histopathological examination of rats as indictors to systematically study the synergistic effect of suet oil in warming kidney and enhancing yang of fried Epimedii Folium in the dosage form of self-assembled micelles. Drugs were given orally, and compared with model group, indicating normal physical signs and biochemical indicators, and significant alleviation in disorder of hypothalamic-pituitary-adrenal-thymus (HPAT) axis suppression. All these experimental results indicated that both Epimedium flavonoids and its self-assembled micelles, especially the processed self-assembled micelles of Epimedii Folium, showed the significant effects in warming kidney and enhancing yang, which might be correlated with the regulation of the disorder of HPAT axis suppression and the improvement of major pathological conditions of testicular gonadal tissues. This discovery can further reveal the synergistic effect and mechanism of processing material suet oil on Epimedii Folium.

Relationship between hepatic drug-metabolizing enzymes CYP450 and traditional Chinese medicine-induced hepatotoxicity / 中国中药杂志

In recent years, with the emergence of new methods and technologies in traditional Chinese medicines metabolism, the relationship between medicine metabolism and cytochrome P450 has gradually been revealed. The research on P450 drug metabolizing enzymes can be used to predict the side effects of traditional Chinese medicines and explore the relationship between compatibility of medicines and toxicity reducing and efficacy enhancing. This paper aims to summarize the progress of CYP450 research, the mechanism of hepatic drug-metabolizing enzymes in the process of drug-metabolism and the relationship between CYP450 and medicine hepatotoxicity. Furthermore, we set out the regulation effects of typical traditional Chinese medicines on CYP450 to provide a reliable basis for the rational use of Chinese medicines.

Study on totai flavonoids of Epimedium assisted with soybean polysaccharide spray-drying powder / 中国中药杂志

In order to evaluate the characteristics of the spray drying of total flavonoids of Epimedium extracts assisted with soybean polysaccharide, a certain percentage of soybean polysaccharide or polyvidone were added to the total flavonoids of Epimedium extract to conduct the spray drying. The effect of soybean polysaccharides against the wall sticking effect of the spray drying was detected, as well as the powder property of total flavonoids of Epimedium spray drying powder and the dissolution in vitro behavior of the effective component. Compared with the total flavonoids of Epimedium spray drying powder, soybean polysaccharide revealed a significant anti-wall sticking effect. The spray drying power which had no notable change in the grain size made a increase in the fluidity, improvement in the moisture absorption and remarkable rise in the dissolution in vitro behavior. It was worth further studying the application of soybean polysaccharide in spray drying power of traditional Chinese medicine.

Study on porous starch as solid dispersion carrier of total Epimedium flavonoids / 中国中药杂志

In order to evaluate the characteristic of porous starch (PS) as the solid dispersions carrier of the total Epimedium flavonoids (TEF), the PS was used. The dissolution of icariin was selected as an indicator to analyze the differences of dissolution between TEF and its solid dispersion. TEF was characterized by scanning electron microscopy (SEM), differential scanning calorimetry (DSC) and X-ray powder diffraction (XRD). Solid dispersion was irregular block and no powder characteristics of TEF and PS could be seen in SEM, DSC and XRD analysis suggested that TEF may be present in solid dispersion as amorphous substance. The dissolution rate of icariin has been improved significantly when the proportion of TEF and PS was 1:2. PS as a traditional solid dispersion carrier is worthy of further study.

Study on solid dispersion of precipitated calcium carbonate-based oleanolic acid / 中国中药杂志

Oleanolic acid-precipitated calcium carbonate solid dispersion was prepared by using solvent evaporation method. The microscopic structure and physicochemical properties of solid dispersion were analyzed using differential scanning calorimetry and scanning electron microscopy (SEM). And its in vitro release also was investigated. The properties of the precipitated calcium carbonate was studied which was as a carrier of oleanolic acid solid dispersion. Differential scanning calorimetry analysis suggested that oleanolic acid may be present in solid dispersion as amorphous substance. The in vitro release determination results of oleanolic acid-precipitated calcium carbonate (1: 5) solid dispersion showed accumulated dissolution rate of.oleanolic acid was up to 90% at 45 min. Accelerating experiment showed that content and in vitro dissolution of oleanolic acid solid dispersion did not change after storing over 6 months. The results indicated that in vitro dissolution of oleanolic acid was improved greatly by the solid dispersion with precipitated calcium carbonate as a carrier. The solid dispersion is a stabilizing system which has actual applied value.

Study on sustained release preparations of Epimedium component / 中国中药杂志

The formulation for sustained release tablet of Epinedium component was selected and the evaluation equation of in vitro release was established. The liquidity of component was improved with the help of colloidal silica aided by spray drying, which would be the main drug in the sustained release tablets. Dissolution was selected as an evaluation index to investigate skeletal material type, fillers, impact porogen, lubricants and other materials on the quality of sustained release tablet. The sustained release tablets were prepared by dry compression. Formulation of sustained release preparations was main drug 35%, HPMC K(4M) 20% and HPMC K(15M) 10% as skeleton material, MCC 31% as filler, PEG6000 2% as porogen and magnesium stearate 2% as lubricant. The sustained release tablets released up to 80% in 8 h. The zero order equation, primary equation and Higuchi equation could simulate the release characteristics of sustained release tablets in vitro, the correlation coefficients r were larger than 0.96. The primary equation was most similar in vitro release characteristics and its correlation coefficient r was 0.9950. The preparation method is simple and the results of formulation selection are reliable. It can be used to guide the production of Epimedium component sustained release preparations.

Solidification of volatile oil with graphene oxide / 药学学报

To evaluate the properties of solidifying volatile oil with graphene oxide, clove oil and zedoary turmeric oil were solidified by graphene oxide. The amount of graphene oxide was optimized with the eugenol yield and curcumol yield as criteria. Curing powder was characterized by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). The effects of graphene oxide on dissolution in vitro and thermal stability of active components were studied. The optimum solidification ratio of graphene oxide to volatile oil was 1:1. Dissolution rate of active components had rare influence while their thermal stability improved after volatile oil was solidified. Solidifying herbal volatile oil with graphene oxide deserves further study.

Two-dimensional zebrafish model combined with hyphenated chromatographic techniques for evaluation anti-osteoporosis activity of epimendin A and its metabolite baohuoside I / 药学学报

This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.

Ideas and methods of two-dimensional zebrafish model combined with chromatographic techniques in high-throughput screening of active anti-osteoporosis components of traditional Chinese medicines / 中国中药杂志

<p><b>OBJECTIVE</b>To break through the restrictions of the evaluation model and the quantity of compounds by using the two-dimensional zebrafish model combined with chromatographic techniques, and establish a new method for the high-throughput screening of active anti-osteoporosis components.</p><p><b>METHOD</b>According to the research group-related studies and relevant foreign literatures, on the basis of the fact that the zebrafish osteoporosis model could efficiently evaluate the activity, the zebrafish metabolism model could efficiently enrich metabolites and the chromatographic techniques could efficiently separate and analyze components of traditional Chinese medicines, we proposed that the inherent combination of the three methods is expected to efficiently decode in vivo and in vitro efficacious anti-osteoporosis materials of traditional Chinese medicines.</p><p><b>RESULT AND CONCLUSION</b>The method makes it simple and efficient in the enrichment, separation and analysis on components of traditional Chinese medicines, particularly micro-components and metabolites and the screening anti-osteoporosis activity, fully reflects that efficacious materials of traditional Chinese medicines contain original components and metabolites, with characteristic of "multi-components, multi-targets and integral effect", which provides new ideas and methods for the early and rapid discovery of active anti-osteoporosis components of traditional Chinese medicines.</p>

Effects of micronization on micromeritics properties of baicalin / 中国中药杂志

Baicalin extremely fine powder was made by using ball-mill and the effect of micronization on the micromeritics properties of baicalin was studied and analyzed. The microstructures of baicalin ordinary and extremely fine powder were compared by scanning electron microscope, differential scanning calorimeter and X-ray diffraction and the powder characteristic of them was investigated. The hygroscopicity was studied. The effect of micronization on the dissolution of baicalin was investigated. The results showed that the chemical constituents of baicalin were not changed after micronization with better compressibility. It was confirmed that micronization technology had a certain application value in promoting the insoluble component of baicalin absorption with higher dissolution.

Study on oral absorption enhancers of astragalus polysaccharides / 中国中药杂志

Astragalus polysaccharides was lounded to 4-(2-aminoethylphenol), followed by labeling the APS-Tyr with fluorescein-5-isothiocyanate (FITC) at the secondary amino group. The absorption enhancement effects of low molecular weight chitosan and protamine on astragalus polysaccharides were evaluated via Caco-2 cell culture model. The results show that the fluorecent labeling compound has good stability and high sensitivity. On the other hand low molecular weight chitosan and protamine also can promoted absorption of the astragalus polysaccharides without any cytotoxity, and the absorption increase was more significant with increasing the amount of low molecular weight chitosan and protamine. At the same time, the low molecular weight chitosan has slightly better effect. The transepithelial electric resistance (TEER) of Caco-2 cells show that absorption enhancers could improve its membrane transport permeability by opening tight junctions between cells and increasing the cell membrane fluidity.