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Establish a production line with controllable process and high intelligence, contribute to improve the quality and production efficiency of aconite processed by microwave, and promote the transformation and application of aconite processed by microwave. According to the principle of aconite detoxification and the characteristics of industrial microwave equipment, an industrial production line of aconite processed by microwave was established with diester alkaloids and monoester alkaloids as indicators, and pilot production was carried out. At the same time, the content of active constituents and efficacy were compared with that of the main processed products, such as Shengfupian, Baifupian and Heishunpian. The results showed that the industrial production of aconite processed by microwave can be divided into two stages: "Liquid seal to detoxification - drying and puffing". The content of monoester alkaloids in 10 batches of aconite processed by microwave was 0.071%-0.166% and the content of diester alkaloids was 0.004%-0.016%, which met the relevant requirements of the Chinese Pharmacopoeia in 2020. Compared with Heishunpian and Baifupian, the retention rate of the effective components of aconite processed by microwave was higher. Pharmacological experiments showed that aconite processed by microwave not only retained the anti-inflammatory and analgesic activities of Heishunpian and Baifupian, but also significantly increased the levels of leukocytes and lymphocytes in mice with liver cancer chemotherapy, enhanced the CD4/CD8 ratio in spleen cells of mice (P < 0.05), thus regulating the body's immunity. However, this effect of Baifupian was weak, while Heishunpian and Shengfupian had no such effect. Through the above research, this study established microwave processing line with controllable process and high intelligence, as well produced the aconite processed by microwave with low toxicity and stable quality. It laid a foundation for the industrialized continuous production and clinical positioning of aconite by microwave processed, and provided scientific support for the development and application of microwave technology in the field of traditional Chinese medicine. All animal experiments in this study were reviewed and approved by the Experimental Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine before being carried out (Approval No. 2020-28).
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ObjectiveTo investigate the effect of modified Renshen Wumeitang(MRWT) on the related regulatory factors of the γ-aminobutyric acid (GABA) signaling pathway in colon tissues of rats with diarrhea, and reveal the mechanism of MRWT in invigorating Qi, generating fluid, and checking diarrhea. MethodForty-eight SD immature rats were randomly divided into a blank group (n=12) and an experimental group (n=36). The diarrhea model was induced in the experimental group by Sennae Folium combined with overstrain and improper diet for 14 days. Subsequently, the model rats were randomly divided into a model group (normal saline, 20 mL·kg-1), a western medicine group (Medilac-Vita, 0.7 g·kg-1), and a Chinese medicine group (MRWT, 35 g·kg-1), with 12 rats in each group. The rats in the blank group received normal saline at 20 mL·kg-1, and those in the other groups were treated correspondingly, once a day for 7 days. The general condition, loose stool rate, and diarrhea index of the rats were observed daily. Immunohistochemistry was used to detect the optical density expression of GABA protein in the colon of rats. The content of phosphatidylinositol-3 kinase (PI3K), protein kinase B2 (Akt2), phosphorylated Akt (p-Akt), and interleukin-1β (IL-1β) was determined by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression levels of PI3K, Akt2, and GABA type A receptor subunit β2 (GABRB2) in the colon of rats were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. ResultCompared with the blank group, the model group showed worsened general condition, The difference was not statistically significant of loose stool rate and diarrhea index, increased expression of GABA protein (P<0.05), elevated expression of PI3K, Akt2, p-Akt, and IL-1β (P<0.05, P<0.01), and up-regulated PI3K, Akt2, and GABRB2 mRNA and protein expression (P<0.01). Compared with the model group, the western medicine group and the Chinese medicine group showed the improved general condition, decreased loose stool rate and diarrhea index (P<0.01), and decreased content of PI3K, Akt2, p-Akt, and IL-1β (P<0.05). The Chinese medicine group displayed decreased mRNA expression of PI3K, Akt2, and GABRB2 (P<0.05, P<0.01) and down-regulated protein expression of GABA, PI3K, and GABRB2 (P<0.05, P<0.01). The western medicine group exhibited down-regulated mRNA expression of PI3K,Akt2,and protein of PI3K (P<0.05). ConclusionMRWT can regulate the GABA signaling pathway, reduce Cl- flow in intestinal epithelial cells to the intestinal lumen, and improve the imbalance of colonic fluid metabolism in the colon of diarrhea rats, thereby exerting its effects of invigorating qi, generating fluid, and checking diarrhea.
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The processing of Chinese herbal medicine is a form of pharmaceutical technology developed over thousands of years, in order to increase efficiency and decrease toxicity of herbs in traditional Chinese medicine (TCM). Herbal processing is essential for safe and effective application of TCM in clinical practice, as it alters the active chemical components and therefore the functions of herbal medicines. Alkaloid-rich herbal medicines in TCM are commonly processed by cleansing, cutting, processing by dry stir-frying, stir-frying with liquid adjuvants, and processing by water decoction. In addition, commonly used adjuvants for processing alkaloid-rich herbal medicines are river sand, wine, vinegar, brine, honey and herbal juice. For alkaloid-rich herbal medicines, the main chemical reactions that occur during processing include hydrolysis, oxidation, replacement, decomposition and condensation. This paper aimed to summarize the processing methods and mechanisms for alkaloid-rich Chinese herbal medicines, and provide much-needed theoretical support and scientific evidence for understanding those mechanisms and effects. Information on processing methods for alkaloid-rich herbal medicines was collected from classic books of herbal medicine, PhD and MSc dissertations, online scientific databases including PubMed, SciFinder, Scopus, Web of Science, Baidu Scholar and Google Scholar. This paper should help to advance our knowledge of the processing mechanisms and aid in the development of processing methods for alkaloid-rich Chinese herbal medicines.
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Objective:To investigate the therapeutic effect of Guizhi Shaoyao Zhimutang (GSZT) on the osteoporosis (OP) in SD female rats induced by ovariectomy (OVX) combined with glucocorticoid injection and its related mechanisms. Method:The rats were divided into a sham operation group and an experimental group for OVX. One week later, the experimental rats were divided into an OP model group, a calciferol (positive control, 0.1 mg·kg<sup>-1</sup>) group, and low(0.8 g·kg<sup>-1</sup>), medium(1.6 g·kg<sup>-1</sup>), and high-dose (3.2 g·kg<sup>-1</sup>) GSZT groups. Except for those in the sham operation group, the rats received an intramuscular injection of dexamethasone (1 mg·kg<sup>-1</sup>) twice per week for six weeks. After the rats were treated correspondingly for eight weeks, the rats were sacrificed for thighbone sample collection. The computer microtomography (Micro CT) was used to analyze the parameters of bone mineral density (BMD), bone volume (BV), tissue volume (TV), structure model index (SMI), connectivity density (Conn.D), trabecular number (Tb.N), trabecular thickness (Tb.Th), trabecular pattern factor (Tb.Pf), and trabecular separation (Tb.Sp). In addition, hematoxylin-eosin (HE) and Masson staining was used for the histopathological examination of the thighbone. Tartrate-resistant acid phosphatase (TRAP) staining was used to analyze osteoclasts in bone tissues. The enzyme-linked immunosorbent assay (ELISA) was used to determine matrix metalloproteinase-9 (MMP-9), cathepsin K (CTSK), and TRAP5b in bone tissues, and Western blot was carried out to determine receptor activator of NF-kappaB ligand (RANKL), osteoprotegerin (OPG), alkaline phosphatase (ALP) in bone tissues. Result:Compared with normal group, the number of bone trabeculae in cancellous part of OP model rats decreased significantly, the spacing widened, the thickness became thinner, and the continuity was poor. Compared with model group, GZST medium and high-dose groups could improve the changes of femoral bone trabecular cancellous bone. Masson staining showed that compared with the normal group, the growth plate thickness of the distal femur epiphysis was thinner and the area of new bone was smaller in the OP model group, while the reduction of new bone was inhibited in the high dose group of calcitol and GZST. TRAP staining results showed that the number of osteoclasts in OP group was significantly increased compared with the normal group, and the number of osteoclasts in medium-high dose GZST group was reduced. Micro CT results showed that compared with normal group, BMD, BV, Tb.N, Tb.Th and Conn.D of OP rats were decreased, SMI and Tb.Sp were significantly increased(<italic>P</italic><0.01). Compared with model group, medium and high dose of GZST could improve the degree of osteoporosis in OP rats, BV, Tb.N were significantly increased, SMI, Tb.Sp were significantly decreased (<italic>P</italic><0.01). ELISA results revealed that compared with the sham operation group, the OP model group showed decreased ALP content (<italic>P</italic><0.01) and increased MMP-9, TRAP5b, and CTSK (<italic>P</italic><0.01). Compared with the OP model group, the high-dose GSZT group showed increased ALP content (<italic>P</italic><0.01) and decreased MMP-9, TRAP5b, and CTSK (<italic>P</italic><0.01). Western blot results demonstrated that compared with the OP model group, the calciferol group and the high-dose GSZT group showed inhibited expression of the RANKL protein (<italic>P</italic><0.05), and increased OPG expression (<italic>P</italic><0.05). Conclusion:The findings suggest that GZST has a potential therapeutic effect on OP caused by OVX combined with glucocorticoids in rats, and the underlying mechanism is associated with the regulation of RANKL/OPG in bone tissues. This study is expected to lay a preliminary foundation for its clinical application and modern preparation development.
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Objective:To explore the material basis and mechanism of Sangjiang Ganmao injection (SG) in the treatment of common cold by ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) and network pharmacology. Method:UPLC-Q-Orbitrap HRMS was used to identify the chemical components of SG with mobile phase of acetonitrile (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-10 min, 4%-15%A; 10-35 min, 15%-30%A; 35-45 min, 30%-33%A; 45-55 min, 33%-60%A; 55-58 min, 60%A), flow rate of 0.2 mL·min<sup>-1</sup>, electrospray ionization (ESI) and scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500 under positive and negative ion modes. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and GeneCards 5.0 database were used to screen and predict the potential targets of chemical components in SG, STRING 11.0 database and Cytoscape 3.7.2 software were used to construct protein-protein interaction (PPI) network model, gene ontology (GO) analysis and pathway analysis were performed on potential targets by Metascape 3.5, Reactome database and Kyoto Encyclopedia of Genes and Genomes (KEGG), Cytoscape 3.7.2 software was used to build the network of "herbs-ingredients-key targets". Result:A total of 54 components in SG were identified, and 80 potential targets of SG for treatment of common cold were predicted and screened based on this. SG exerted therapeutic effects by acting on targets such as interleukin (IL)-6, tumor necrosis factor (TNF) and IL-10, and signaling pathways such as IL-17 signaling pathway, TNF signaling pathway and interaction of cytokine receptors. Conclusion:SG may interfere with the expression of inflammatory cytokines by acting on related targets and pathways such as inflammation and immune system, and regulate the immune function of the body as a whole, thereby exerting a therapeutic effect.
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Objective:Based on UPLC-Q-Orbitrap HRMS and network pharmacology, the material basis, processing principle and molecular mechanism of bile processed Coptidis Rhizoma (BPRC) for reducing excess fire of liver and gallbladder were elucidated.Method:The chemical ingredients of BPRC were analyzed by UPLC-Q-Orbitrap HRMS. Chromatographic separation was achieved with 0.1% formic acid solution (A)-acetonitrile (B) as the mobile phase in gradient elution (0-20 min, 5%-80%B; 20-30 min, 80%-95%B; 30-30.1 min, 95%-5%B; 30.1-35 min, 95%-5%B). The flow rate was 0.2 mL·min-1, electrospray ionization (ESI) was applied and operated in positive and negative ion modes, the acquisition range was m/z 100-1 500. Based on the clinical manifestations and pathogenic factors of excess fire of liver and gallbladder, the potential effective ingredients, targets and functional characteristics of BPRC were predicted and analyzed by online database. Based on the characteristics of the new active ingredients after processing, the processing principle of BPRC was investigated by network pharmacology.Result:A total of 19 ingredients in BPRC were identified, six of which were newly added cholic acids after processing. It was determined that the alkaloids, including worenine, epiberberine, jatrorrhizine, coptisine, berberrubine, berberine, palmatine and cholic acids, including glycohyodeoxycholic acid, taurohyodeoxycholic acid, glycochenodeoxycholic acid, hyodeoxycholic acid and taurochenodeoxycholic acid, were identified as material basis of BPRC. A total of 66 targets of reducing excess fire of liver and gallbladder of BPRC were screened. There were 16 common targets and multiple same signaling pathways between cholic acids and alkaloids of BPRC, and many lesions of excess fire of liver and gallbladder were target organs of cholic acids. By acting on some targets, including albumin (ALB), Caspase-3 (CASP3), mitogen-activated protein kinase 14 (MAPK14), glucocorticoid receptor (NR3C1) and other targets and some signaling pathways, including interleukin (IL)-17, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), MAPK and other pathways, BPRC could reduce excess fire of liver and gallbladder.Conclusion:BPRC has the characteristics of multi-component, multi-target and multi-pathway on reducing excess fire of liver and gallbladder. Bile and Coptidis Rhizoma have synergistic effect and bile can enhance the intensity of BPRC in lesions, which confirms the processing theory that the effect of BPRC on excess fire of liver and gallbladder enhance after being processed by bile.
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Objective: To investigate the effect of Coptidis Rhizoma on intestinal microflora of normal and type 2 diabetic rats before and after being steamed with rice wine based on 16S rRNA sequencing technology. Method: The type 2 diabetic rat model was established by high-fat diet and streptozotocin. Normal and model rats were given the decoction of Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine every day (0.8 g·kg-1, according to raw drug quantity), the positive drug group was given metformin solution every day (0.25 g·kg-1), continuous administration for 30 d. Blood glucose was measured once a week and oral glucose tolerance was measured on the 27th day of administration. On the 30th day, feces were taken and the rats were anesthetized. Blood was taken for enzyme linked immunosorbent assay (ELISA). Colon was dissected for hematoxylin-eosin (HE) staining. Based on 16S rRNA gene sequencing technology, the alpha diversity, beta diversity, abundance and composition, and the correlation with anti-inflammatory and antioxidant activities of intestinal microflora were comprehensively analyzed and evaluated. Result: After giving normal rats Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine, there were some inflammatory reactions, imbalance of oxidation-antioxidant system, pathological damage of colon and changes of intestinal microflora, which showed some toxic and side effects, but the toxic and side effects of Coptidis Rhizoma steamed with rice wine were lower than those of Coptidis Rhizoma. Coptidis Rhizoma could reduce fasting blood glucose and improve glucose tolerance in type 2 diabetic rats before and after steaming. Although the effect of Coptidis Rhizoma steamed with rice wine had an increasing trend, there was no significant difference between before and after processing. Type 2 diabetic rats appeared mild inflammation and imbalance of oxidation-antioxidant system, characterized by significant increase of contents of interleukin (IL)-6, nuclear transcription factor (NF)-κB and malondialdehyde (MDA) (PPPConclusion: Coptidis Rhizoma can treat type 2 diabetic rats by regulating intestinal microflora before and after steaming with rice wine, and the effect of Coptidis Rhizoma steamed with rice wine is better. The toxic and side effects of Coptidis Rhizoma on intestinal flora are reduced by steamed with rice wine. The processing mechanism of increasing efficacy and decreasing toxicity of Coptidis Rhizoma steamed with rice wine is closely related to intestinal microflora.
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To establish the spectrum-effect relationship between HPLC fingerprint and free radicals activity scavenging in Guizhi Shaoyao Zhimu Decoction( GSZD),and provide a basis for the quality evaluation and modernization of classical prescriptions. Shimadsu GL-science C18 column( 4. 6 mm×250 mm,5 μm) was used with acetonitrile-0. 1% formic acid solution as the mobile phase for gradient elution. The detective wave length was 254 nm; the column temperature was set at 32 ℃; the injection volume was 20 μL; and the flow rate was 1. 0 m L·min-1.10 batches of primary standard samples of GSZD were detected,and their HPLC fingerprint was established by using the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine( TCM). The activity of scavenging free radicals was studied by 1,1-diphenyl-2-trinitrophenylhydrazine( DPPH) method,and the spectrum-effect relationship was studied by Pearson bivariate correlation analysis. The common mode of GSZD fingerprints was established,and 26 common peaks were marked,with similarities ranging from 0. 929 to 0. 998. Eight of the chromatographic peaks were identified by using the control comparison method: gallic acid,mangiferin,paeoniflorin,glycyrrhizin,asparagus,5-O-methylvisamicin,cinnamic acid,and ammonium glycyrrhetate. Among them,the content changes of No. 14( paeoniside),20,12( mangiferin),13 and 23( cinnamic acid) common peaks were negatively correlated with free radical scavenging activity. The fingerprint showed high precision,repeatability and stability,and the common peaks were well separated,so it can be used for the quality evaluation of GSZD,and could provide reference for further studies on the material basis of GSZD.
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Chromatography, High Pressure Liquid , Cinnamomum aromaticum , Chemistry , Drugs, Chinese Herbal , Chemistry , Free Radical Scavengers , Chemistry , Medicine, Chinese TraditionalABSTRACT
To study the variation of six ester-type alkaloids and characteristic fingerprints in the process from Radix Aconite Lateralis to Heishunpian and lay a foundation for the study of the processing principle of Heishunpian, HPLC. analysis was performed on a Phenomenex Gemini C18 (4.6 mm x 250 mm, 5 microm) with acetonitrile and 40 mmol x L(-1) ammonium acetate (adjusted to pH 10 with concentrated ammonia water) as mobile phase. The detection wavelength was set at 235 nm. The flow rate was set at 0.8 mL x min(-1) and the injection volume was 10-20 microL. Six ester-type alkaloids were determined and characteristic fingerprints of the process were established. As the process continues, the contents of diester diterpene alkaloids were decreased step by step, while the contents varia tion of monoester diterpene alkaloids were not obvious. Each sample showed significant difference in characteristic fingerprints. With the exception of 6 known monoester diterpene alkaloids and diester diterpene alkaloids, 13 peaks were marked in the characteristic fingerprints, of which the total change rule of the other 7 unknown peaks were similar with 3 diester diterpene alkaloids. The established method is accurate, reliable and repeatable, and can provide reference for revealing change rule of index components and illuminating processing principle in the process of Heishunpian.
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Aconitine , Chemistry , Aconitum , Chemistry , Alkaloids , Chemistry , Chemistry, Pharmaceutical , Methods , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Esters , Chemistry , Molecular StructureABSTRACT
Different processed products of Coptidis Rhizoma have its unique odor, which is an important assessment index for pro- cessed products identification of Coptidis Rhizoma. Objectify odor as an entry point in this study, an electronic nose technology was used, and a suitable method for Coptidis Rhizoma measurement was built firstly. Then different processed products of Coptidis Rhizoma were detected by the method built. Finally, different processed products were identified by combining with chemometrics based on the objective odor information obtained. Electronic nose detection indicated that a significant difference in odor between different processed products was performed. Coptidis Rhizoma processed or not can be distinguished based on statistical quality control (SQC) and soft independent modeling of class analogy (SIMCA). Principle component analysis (PCA) model showed that Coptidis Rhizoma and its various processed products discriminated obviously. In addition, in order to identify the processed products of Coptidis Rhizoma, a correct recognition rate of 100% was acquired by discriminant factor analysis (DFA) , and the initial identification rate and cross-validation recognition rate of linear discriminant analysis (LDA) is 100%, 94.4% respectively. In conclusion, differentiationin odor of different processed Coptidis Rhizoma was performed by the electronic nose technology used, and different products Coptidis Rhizoma were dis- criminated by combining with chemometrics. This research can be a reference for objective identification in odor of traditional Chinese medicine, and is good for the inheritance and development of traditional experience in odor identification.
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Chemistry, Pharmaceutical , Chromatography, High Pressure Liquid , Coptis , Chemistry , Drugs, Chinese Herbal , Electronic Nose , Odorants , Principal Component Analysis , Rhizome , ChemistryABSTRACT
Dryness is the inherent performance in traditional Chinese medicine. Dryness with a specific efficacy and side effect can be reduced suitably by processing and compatibility in the clinical application. Nowadays domestic scholars have developed research of dryness in traditional Chinese medicine. However, it remains problems such as evaluation index of dryness not clear. This paper takes medical literature mining technology to analyze the historical origin and features of dryness theory. Combing the modern literatures to explicate the dryness' research status and existing problems. Putting forward the traditional Chinese medicine and research should adopt multidisciplinary knowledge and study the system of comprehensive evaluation. Dryness is expected to further application in traditional Chinese medicine clinical research.
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Humans , Data Mining , Drug Compounding , Methods , Medicine, Chinese Traditional , ResearchABSTRACT
<p><b>OBJECTIVE</b>Combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, its aims to provide experimental basis for the classification and quality evaluation of Coptis chinensis.</p><p><b>METHOD</b>Fourteen batches of C. chinensis from different areas were respectively measured in size (total length, total width, root length, taproot diameter, branch number, branch length, branch diameter, length of the bridge, weight), color (external color, internal color), content (epiberberine, coptisine, palmatine, berberine). Then the determination data were evaluated by spss principal component analysis and cluster analysis.</p><p><b>RESULT</b>Three principal components were extracted from the original data. The principal component analysis results showed that the characteristic elements might be the total length, main root length, taproot diameter, branch length, weight, the total color value of the appearance and content of epiberberine and berberine. The results of cluster analysis showed that 14 batches of samples could be clustered reasonably into two groups. In terms of the appearance and quality, there were some differences between in the geo-authentic and non-authentic producing areas of C. chinensis.</p><p><b>CONCLUSION</b>The method which was combining the quantitative physical property characteristics of the appearance with the internal quality evaluation index, and through the processing of mathematical statistics, could be used for the the classification of C. chinensis.</p>