Comparative pharmacokinetics of multi-components in normal and stomach cold syndrome rats after oral administration of Zingiberis Rhizoma - Jujubae Fructus herb pair and its single herb extracts by UHPLC-MS/MS.

The Zingiberis Rhizoma - Jujubae Fructus herb pair (ZJHP) is a classic herb pair in traditional Chinese medicine. The herb pair shows the effect of dispelling cold, harmonizing the middle and improving gastrointestinal function, and is widely used for patients with stomach cold syndrome (SCS), stomachache and anemofrigid cold. The gingerols, shogaols, flavonoids and triterpenic acids are the important bioactive ingredients of ZJHP. However, few pharmacokinetic studies have been investigated in vivo for the above compounds. To comprehend the kinetics of active components and promote their curative application, a fast and sensitive ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) method was established for simultaneous determination of 12 analytes in normal and SCS rats in this study. The results showed that the pharmacokinetic parameters (Cmax, Tmax, t1/2z, MRT0-t, AUC0-t and AUC0-∞) in SCS model were significantly different from those in normal rats. In addition, the pharmacokinetics of rats given ZJHP were also varied from single herb oral administration, especially in model condition. These results indicated that the in vivo processes of the above analytes changed under pathological conditions and the compatibility of the herb pair could significantly influence the absorption of active components, which might provide an insight and further supports for the clinical application of ZJHP.

Dimeric diarylheptanoids with anti-inflammatory activity from Zingiber officinale.

Two previously undescribed chain diarylheptanoid derivatives (2-3), five previously undescribed dimeric diarylheptanoids (4-8), together with one known cyclic diarylheptanoid (1) were isolated from Zingiber officinale. Their structures were elucidated by extensive spectroscopic analyses (HR-ESI-MS, IR, UV, 1D and 2D NMR) and ECD calculations. Biological evaluation of compounds 1-8 revealed that compounds 2, 3 and 4 could inhibit nitrite oxide and IL-6 production in lipopolysaccharide induced RAW264.7 cells in a dose-dependent manner.

Protective Effect of Zingiberis Rhizoma Recens on LPS-induced Acute Lung Injury in Mice Through TLR4/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the protective effect and mechanism of Zingiberis Rhizoma Recens alcohol extract on lipopolysaccharide （LPS）-induced acute lung injury in mice. MethodBalb/c mice were randomly divided into normal group， model group， dexamethasone group， and low- and high-dose Zingiberis Rhizoma Recens groups. Mice in the normal group were instilled with normal saline through the nose， and the other groups were instilled with normal saline containing LPS （50 μg）. After 30 minutes of modeling， the dexamethasone group was gavaged with 5 mg·kg-1 of dexamethasone acetate solution， the low- and high-dose Zingiberis Rhizoma Recens groups were gavaged with different doses of （7， 14 g·kg-1） of Zingiberis Rhizoma Recens alcohol extract， and the normal group and the model group were gavaged with the same volume of water. After 24 hours of modeling， the total number of white blood cells in bronchoalceolar lavage fluid （BALF） was detected by cell counter， and the levels of the inflammatory factors including tumor necrosis factor （TNF）-α， interleukin （IL）-1β， IL-6， and superoxide dismutase （SOD）， and myeloperoxidase （MPO） was detected by enzyme-linked immunosorbent assay （ELISA）. Haematoxylin-eosin （HE） staining method was used to observe the pathological changes of lung tissue in each group， and the Western blot was used to detect the protein expression of nuclear transcription factor （NF）-κB p65， phosphorylation （p）-NF-κB p65， and Toll-like receptor 4 （TLR4） in lung tissue. ResultCompared with the normal group， the white blood cell count in BALF and the levels of TNF-α， IL-1β， IL-6， and MPO in the model group was increased （P<0.01）， and the level of SOD was decreased （P<0.05）. Pathological damage of lung tissue was obvious， and the protein expression of NF-κB p65， p-NF-κB p65， and TLR4 in lung tissue was increased （P<0.01）. Compared with the model group， the white blood cell count in BALF and the levels of TNF-α， IL-1β， IL-6， and MPO in the treatment group was decreased （P<0.05，P<0.01）， and the level of SOD was increased （P<0.05，P<0.01）. Pathological damage of lung tissue was alleviated， and the protein expression of NF-κB p65， p-NF-κB p65， and TLR4 in lung tissue was decreased （P<0.01）. ConclusionZingiberis Rhizoma Recens alcohol extract may play a protective role in LPS-induced acute lung injury in mice by inhibiting the TLR4/NF-κB signaling pathway.

Using ultra-performance liquid chromatography with linear ion trap-electrostatic field orbitrap mass spectrometry, network pharmacology, and molecular docking to explore the constituent targets and action mechanisms of decoction of Angelica sinensis, Zingiberis Rhizoma Recens, and Mutton in the treatment of diarrhea-predominant irritable bowel syndrome.

BACKGROUND: Irritable bowel syndrome (IBS) is a chronic intestinal disorder characterized by abdominal discomfort, stool characteristics, and changes in bowel habits. Among them, diarrhea-type (diarrhea-predominant irritable bowel syndrome, abbreviated as IBS-D) is the most common. Because its pathogenesis is not understood, symptomatic treatment is currently used in clinical practice, and its long-term effect is still unclear. Decoction of Angelica sinensis, Zingiberis Rhizoma Recens, and Mutton (DAZM) is a famous traditional Chinese medicine recipe created by Zhang Zhongjing, a famous doctor in the Eastern Han Dynasty 2000 years ago, and is still in use today. Our research team has previously investigated the clinical study of DAZM in the treatment of IBS-D and conducted animal experiment research, indicating that DAZM has a significant effect on improving IBS-D. Yet, there are few reports on the specific mechanism of action of DAZM in improving the treatment of IBS and related types. Most studies discuss and verify its efficacy and protection from a clinical perspective. For this reason, this research will explore the constituent targets and mechanisms of DAZM to improve the treatment of IBS-D, provide relevant scientific evidence, and also provide reference evidence for the efficacy of food therapy decoction in improving the treatment of diseases and mechanism to open up new experimental research ideas. METHODS: Identification of drug ingredients and collection of targets for DAZM using ultra-performance liquid chromatography with linear ion trap-electrostatic field orbitrap mass spectrometry and the Bioinformatics Analysis Tool for Molecular Mechanisms of Traditional Chinese Medicine database, active ingredients were selected based on their oral bioavailability and drug-like properties. Obtained IBS-D targets using the GeneCards database, took the intersection of IBS-D targets and DAZM targets and obtained potential targets of DAZM for the treatment of IBS-D. Using Cytoscape software to draw a network diagram of "Food therapy decoction-ingredient-target-disease" and selected the ingredients with larger parameter values by topological analysis. Correlation analysis of the selected active and parametric ingredients with prominent symptoms of IBS-D using SymMap database, and selection of potential core ingredients. The construction of protein interaction networks from the String database and the selection of potential core targets. Gene Ontology functional and Kyoto Encyclopedia of Genes and Genomes signaling pathway enrichment analyses using the Metascape database, establishing the bioinformatic processes and signaling pathways involved. Molecular docking of core ingredients and potential core targets was performed using AutoDock Vina, and the results were visualized using Python molecule (PyMOL) and LigPlus+. Finally, based on the results of this research combined with previous literature reports, the discussion section of this paper summarizes in detail the key core ingredients, targets, and signaling pathways of DAZM in improving IBS-D. RESULTS: DAZM may act on eight potential core targets (threonine kinase 1, insulin, tumour necrosis factor, tumour protein p53, interleukin 6, epidermal growth factor receptor, connexin ß1, and interleukin 1ß) through eight core ingredients (Zingiberone, Shyobunone, Palmitic acid, Sebiferic acid, ß-Bisabolene, ß-Sitosterol, Stigmasterol, and Oleic acid). inhibit pro-inflammatory factors through Advanced Glycation End Products-Receptor (AGE-RAGE) signaling pathway, Cyclic Adenosine Monophosphate (cAMP) signaling pathway, MAPK signaling pathway, IL-17 signaling pathway, Calmodulin (CaM) signaling pathway, and other pathways. It can alleviate the inflammatory response, enhance the intestinal mucosal barrier and regulate intestinal motility, and play a role in the treatment and improvement of IBS-D. CONCLUSIONS: This research mainly found the mechanism of DAZM on IBS-D, which may involve multiple ingredients, multiple targets, and multiple pathways. DAZM with medicinal and edible functions can effectively improve the treatment of IBS-D. This kind of dietary therapy is suitable for long-term treatment and is worthy of promotion.

Effects of decoction of Angelica Sinensis, Zingiberis Rhizoma Recens, and mutton on physiology and biochemistry of Sprague Dawley female rats with spleen-kidney Yang deficiency.

Background: To examine the effects of each dose of decoction of Angelica sinensis (Dang gui), Zingiberis Rhizoma Recens (Sheng jiang), and mutton (DAZM) on the physiological and biochemical indexes of female rats with spleen-kidney Yang deficiency (SKYD) through 30-day feeding of DAZM, and to evaluate the tonifying effect of DAZM combined with the system of benefit damage index-general score (BDI-GS). Methods: Sprague Dawley (SD) rats were administered adenine and senna water to establish a SKYD model. The rats were then allocated to 4 groups at random: Model group, and L group, 4.2 g/kg, M group, 8.4 g/kg and H group, 16.8 g/kg. In addition, the group of normal feeding with unlimited diet was set as N group. Blood samples were taken to detect the relevant physiological and biochemical indexes. For organ coefficient analysis, 10 kinds of organ tissues were dissected and weighed. The tonifying effect of DAZM was discussed according to the BDI-GS system. Results: During the modeling, the weight of rats in the normal group displayed a marked growth trend, and the weight of the model group was markedly lower than that of the normal group. After feeding the rats DAZM at a low, intermediate, and high dose, the anal temperature of rats in each group continued to rise, and finally remained basically the same as that of normal rats. Hematological and urine examinations revealed that the urea nitrogen and creatinine (CRE) of the model group and the experimental group were markedly higher than those of the normal group, and there were marked differences. After intragastric administration of DAZM, E2 increased markedly. The BDI-GS values of the liver, spleen, lung, kidney, brain, ovary, and adrenal gland of female rats in the 3 administration groups of DAZM were >1, and the total cumulative GS value of each organ of female rats was more than 10. Conclusions: The decoction of DAZM has no obvious effect on the growth, metabolism, and development of female rats with SKYD, causes no obvious damage to organs, and has a certain reparative effect on the kidney damage caused by SKYD.

Five new compounds from Zingiberis Rhizoma Recens and their anti-apoptotic activity.

Five new compounds, named gingerol A (1a and 1b), gingerol B (2), diphenylheptane glycoside A (3) and diphenylheptane glycoside B (4), were isolated from the acetone extract of Zingiberis Rhizoma Recens. The structures of new compounds were elucidated on the basis of spectroscopic methods including UV, IR, 1D NMR, 2D NMR and HR-ESI-MS. Compounds 2-4 could significantly decrease the apoptosis rate and increase the survival rate of human normal lung epithelial cells (BEAS-2B) at the concentration of 10 µM.

A network pharmacology integrated pharmacokinetics strategy to investigate the pharmacological mechanism of absorbed components from crude and processed Zingiberis Rhizoma on deficiency-cold and hemorrhagic syndrome.

ETHNOPHARMACOLOGICAL RELEVANCE: Zingiberis Rhizoma (ZR) and Zingiberis Rhizoma Carbonisata (ZRC), as two forms of ginger-based herbal drugs used in China for at least 2000 years, have been recorded in Chinese Pharmacopoeia and applied for specific indications in traditional Chinese medicine (TCM). AIM OF THE STUDY: The present study aimed to explore the underlying therapeutic and processing mechanism of the absorbed components of ZR and ZRC on deficiency-cold and hemorrhagic syndrome (DCHS) using network pharmacological technique combined with pharmacokinetics strategy. MATERIALS AND METHODS: In this study, a rapid and sensitive approach was conceived to simultaneously determine the seven components (zingiberone, 6-gingerol, 8-gingerol, 6-shogaol, 6-paradol, diacetyl-6-gingerol and 10-gingerol) in rat serum by HPLC-DAD-MS. The network pharmacological technique was employed to evaluate the effect of the absorbed components of ZR and ZRC on DCHS. Also, the vitro experiments were carried out to validate the functions of the seven compounds on coagulation and other major haematological effects. RESULTS: The values of intra-assay and inter-assay precision were determined to be less than 7.44%, with an accuracy value ranging from 83.64% to 107.99%. Analysis of rat plasma revealed that the extraction recoveries and matrix effects of the seven analytes were >85.76%. The method for validation following oral administration of ZR and ZRC to rats was proved to be a success in the pharmacokinetic study of the seven ingredients. Pharmacokinetics showed that ZR processing could enhance the absorption and utilization of 6-shogaol, 6-paradol and diacetyl-6-gingerol, meanwhile reduce the absorption of 6-gingerol, 8-gingerol, and 10-gingerol. Through the pathway enrichment analysis, it was found that the significant biological process of ZR and ZRC on DCHS was primarily associated with complement, coagulation cascades and platelet activation pathways. The vitro experiments indicated that zingiberone, 6-paradol and diacetyl-6-gingerol had a hemostatic effect by upregulating the expression of one or more targets such as TNF-α, FⅩa, FⅫ, FⅧ, ICAM-1, vWF and ITGB3. While 6-gingerol, 6-shogaol, 8-gingerol and 10-gingerol played a critical role in promoting blood circulation by increasing the expression of TM and/or PORC, and/or reducing the expression of ITGB3. CONCLUSION: In brief, network pharmacological technique in combination with pharmacokinetics strategy provided an applicable method for pharmacological mechanism study of ZR and ZRC, which, also, could be used as reference for quality control of the two drugs. In a broader sense, this combined strategy might even be valuable in uncovering the therapeutic and processing mechanism of Chinese herbs on a systematic level.

Effects of carbonization processing on quality control, chemical compositions, and pharmacological mechanism of Ganjiang (Zingiberis Rhizoma) / 数字中医药（英文）

@#Ganjiang (Zingiberis Rhizoma, ZR) and Jiangtan (Carbonized Zingiberis Rhizoma, CZR) have long been used in traditional Chinese medicine (TCM) with a rich history in the treatment of various ailments. While ZR and CZR obviously stem from the same botanical source, their attributes, chemical compositions, pharmacological behaviors, and clinical applications are different owing to variations in the extent of drying and processing they undergo. In this paper, data pertaining to ZR and CZR were retrieved from databases including China National Knowledge Infrastructure (CNKI), PubMed, Web of Science, and Google Scholar. These sources were scrutinized to elucidate the distinctions between ZR and CZR arising from carbonization processing in terms of their ethnopharmacology, quality control, chemical compositions, biological activities, pharmacological mechanisms, and clinical uses. In this study, a total of 56 chemical constituents were identified and isolated from ZR and CZR, which primarily encompassed volatile oils, gingerols, and diphenylheptane compounds. CZR's pharmacological effects include hemostatic, anti-oxidant, analgesic, antibacterial, anti-cancer, and other biological activities. ZR has pungent and warm properties. It is a Yang-supplementing herbal medicine for ailments exacerbated by cold or damp climatic influences. CZR is a product of ZR after undergoing high temperature, with diminished intensity of its pungent and warm attributes. This change leads to a more gradual treatment efficacy, renowned hemostatic effects and its ability to gently invigorate the spleen and effectively alleviate diarrhea. Currently, research on the pharmacological mechanism of CZR is mainly focused on the effects of CZR on coagulation and fibrinolysis. Although the healing effect of CZR has long been known, and some correlation has been found between the changing composition and the changing color of the decoctions, people still lack relatively clear processing mechanisms to reflect the characteristics and specific quality standards of the ingredients of CZR's hemostatic effect. This review provides a systematic summary on quality control, chemical composition, ethnopharmacology, and pharmacology of CZR, offering novel perspectives for advancing the exploration of additional carbonized herbal medicine and fostering their application in clinical settings

Metabolomics-based Analysis of Toxicity Changes of Aconiti Lateralis Radix Praeparata-Zingiberis Rhizoma in Sinitang Before and After Compatibility / 中国实验方剂学杂志

ObjectiveTo explore the effect and toxicity change rule of Aconiti Lateralis Radix Praeparata（ALRP） and Zingiberis Rhizoma（ZR） before and after compatibility， and to reveal the compatibility connotation of them. MethodSixty SD rats were randomly divided into blank group， blank-ALRP group， blank-ALRP-ZR group， model group， model-ALRP group and model-ALRP-ZR group， the latter three groups were injected with adriamycin via tail vein to establish the model of heart failure， and the former three groups were injected with the same amount of physiological saline via tail vein. The effects of ALRP single decoction and ALRP-ZR mixed decoction on biochemical indexes and myocardial histopathological morphology of normal rats and model rats were compared. Metabolomics analysis was performed on rat serum samples， principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were used to screen the differential metabolites between groups， and the differential metabolic pathways were analyzed. Combined with network pharmacology technology， the metabolites and their associated targets and pathways related to enhancing anti-heart failure efficacy and reducing cardiotoxicity were screened before and after the compatibility of ALRP and ZR， the screened representative pathways were verified by Western blot. ResultCompared with the blank group， the model group showed significant increases in the contents of brain natriuretic peptide（BNP）， creatine kinase（CK）， lactate dehydrogenase（LDH） and cardiac troponin（cTn）-T（P<0.01）， the blank-ALRP group showed obvious increases in CK， LDH， and cTn-T contents（P<0.05， P<0.01）， while the normal-ALRP-ZR group showed a significant increase in CK content（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed a obvious decrease in LDH content（P<0.05）， and pathological sections showed that both decoctions could lead to myocardial histopathological damage in normal rats. Compared with the model group， the model-ALRP-ZR group showed obvious decreases in BNP， CK， LDH and cTn-T contents（P<0.05， P<0.01）， and the model-ALRP group showed obvious decreases in BNP， LDH and cTn-T contents（P<0.05， P<0.01）. Compared with the model-ALRP group， the model-ALRP-ZR group showed a significant decrease in CK content（P<0.01）， and both decoctions could improve the pathological morphology of myocardial tissue in the model rats. Metabolomics results showed that ALRP single decoction and ALRP-ZR mixed decoction could recover 422 and 459 metabolites in model rats， respectively. And the metabolic disruption of ALRP-ZR mixed decoction on normal rats was weaker than that of ALRP single decoction. The results of network pharmacological association analysis showed that in the aspect of ZR enhancing the anti-heart failure efficacy of ALRP， 3 metabolites such as deoxyuridylic acid were correlated to 56 metabolites， 82 targets and 13 pathways， including calcium signaling pathway， renin secretion， renin-angiotensin system， etc. In the aspect of ZR reducing the cardiotoxicity of ALRP， 3 metabolites such as tyrosol were associated with 24 metabolites， 55 targets and 14 pathways， including adrenergic signaling in cardiomyocytes and carbon metabolism and so on. Western blot results showed that the expression of angiotensin-converting enzyme（ACE）， angiotensin-converting enzyme 2（ACE2） and angiotensin Ⅱ（Ang Ⅱ） in myocardial tissues of rats from the model group was significantly elevated by comparing with the blank group（P<0.01）. Compared with the model group， the model-ALRP group and the model-ALRP-ZR group showed significantly decreased expression of ACE， ACE2 and Ang Ⅱ（P<0.01）. Compared with the model-ALRP group， the expression of ACE2 and AngⅡ was significantly decreased in the model-ALRP-ZR group. Compared with the blank group， the expression of extracellular signal regulated kinase（ERK）， protein kinase B（Akt） and cTn-I3 was significantly elevated in the blank-ALRP group and blank-ALRP-ZR group（P<0.01）. Compared with the blank-ALRP group， the blank-ALRP-ZR group showed decreased expression of ERK， Akt and cTn-I3， but there was no statistical significance. ConclusionTo a certain extent， the combination of ALRP and ZR shows synergistic relationship under pathological state， and attenuated effect of compatibility under normal physiological state， and the pharmacodynamic characteristics and compatibility relationship of ALRP and ZR are closely related to the physiological state.

[Rapid identification of geographic origins of Zingiberis Rhizoma by NIRS combined with chemometrics and machine learning algorithms].

In this study, 280 batches of Zingiberis Rhizoma samples from nine producing areas were analyzed to obtain infrared spectral information based on near-infrared spectroscopy(NIRS). Pluralistic chemometrics such as principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA), orthogonal partial least squares-discriminant analysis(OPLS-DA), K-nearest neighbors(KNN), support vector machine(SVM), random forest(RF), artificial neural network(ANN), and gradient boosting(GB) were applied for tracing of origins. The results showed that the discriminative accuracy of the spectral preprocessing by standard normal variate transformation coupled with the first derivative was 93.9%, which could be used for the construction of the discrimination model. PCA and PLS-DA score plots showed that samples from Shandong, Sichuan, Yunnan, and Guizhou could be effectively distinguished, but the remaining samples were partially overlapped. As revealed by the analysis results by machine learning algorithms, the AUC values of KNN, SVM, RF, ANN, and GB algorithms were 0.96, 0.99, 0.99, 0.99, and 0.98, respectively, with overall prediction accuracies of 83.3%, 89.3%, 90.5%, 91.7%, and 89.3%. It indicated that the developed model was reliable and the machine learning algorithm combined with NIRS for origin identification was sufficiently feasible. OPLS-DA showed that Zingiberis Rhizoma from Sichuan(genuine producing areas) could be significantly distinguished from other regions, with good discriminative accuracy, suggesting that the NIRS established in this study combined with chemometrics can be used for the identification of Zingiberis Rhizoma from Sichuan. This study established a rapid and nondestructive identification and reliable data analysis method for origin identification of Zingiberis Rhizoma, which is expected to provide a new idea for the origin tracing of Chinese medicinal materials.

[Comparative study on intestinal absorption kinetics of main active components in Sini Decoction and its separated recipes].

This paper aims to study the difference in the intestinal absorption kinetics of main active components of Sini decoction and its separated recipes and explain the scientificity and rationality of the compatibility of Sini Decoction. A in situ intestinal perfusion rat model was established to evaluate the differences in the absorption of benzoylmesaconine, benzoylaconine, benzoylhypacoitine, mesaconitine, hypaconitine, glycyrrhizic acid, liquiritin and 6-gingerol from Sini Decoction and its separated recipes in the duodenum, jejunum and ileum by high performance liquid chromatography(HPLC). The results indicated that the Sini Decoction group was superior to the Aconiti Lateralis Radix Praeparata group in terms of absorption degree and rate for aconitum alkaloids. The absorption of benzoylmesaconine and hypaconitine in the duodenum, jejunum and ileum was faster and stronger in the Sini Decoction group(P<0.05). The absorption degree of glycyrrhizic acid in the duodenum was significantly higher in the Sini Decoction group than in the Glycyrrhizae Radix et Rhizoma group and the Glycyrrhizae Radix et Rhizoma-Zingiberis Rhizoma group(P<0.05). The absorption rate and degree of 6-gingerol in the ileum in the Sini Decoction group were significantly higher than those in the Zingiberis Rhizoma group(P<0.05). In short, Zingiberis Rhizoma and Glycyrrhizae Radix et Rhizoma can promote the absorption of aconitum alkaloids in different intestinal segments, which reflects the scientific composition of Sini Decoction.

Integrative analyses of widely targeted metabolomic profiling and derivatization-based LC-MS/MS reveals metabolic changes of Zingiberis Rhizoma and its processed products.

Zingiberis Rhizoma (ZR) has nutritional value and application potentiality, while Zingiberis Rhizoma Praeparatum (ZRP) and Carbonised Ginger (CG) are two main processed products of ZR based on different methods. Here, we performed a widely targeted metabolomics method with Sequential Windowed Acquisition of all Theoretical fragment ions (SWATH) mode to analyze differential metabolites in ZR, ZRP and CG. Additionally, the chemical derivatization was applied to characterize different submetabolomes and improve the separation effect and MS response of metabolites. In total, 369 metabolites were identified and divided into 14 categories, 104 of which were differential metabolites. Our results suggest that carbohydrates, nucleotides, organic acids, vitamins, lipids, indoles, alkaloids, and terpenes contributed to a downward trend after processing, but the maximum content of flavanones, phenylpropanes and polyphenols appeared in ZRP, and that of alcohols appeared in CG. These findings serve as promising perspectives for developing functional food in ZR, ZRP and CG.

[Quality markers of Zingiberis Rhizoma Carbonisata before and after processing].

Based on the previous research results of our group and literature research, the chemical components, mechanisms, pharmacodynamics, and pharmacokinetics of Zingiberis Rhizoma Carbonisata were summarized to determine the quality markers(Q-markers) of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Our research group has clarified the differential components of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma, the meridian-warming hemostatic effect of Zingiberis Rhizoma Carbonisata, the related targets and pathways of the effect, the endogenous biomarkers of Zingiberis Rhizoma Carbonisata, and the hemodynamic processes of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Moreover, based on high-performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry(HPLC-DAD-ESIMS), a method for determining the content of Q-mar-kers was established. In conclusion, the study finally determined that gingerone, 6-shogaol, and diacetyl-6-gingerol were the Q-mar-kers of Zingiberis Rhizoma Carbonisata decoction pieces, and 6-gingerol, 8-gingerol, and 10-gingerol were Q-markers of Zingiberis Rhizoma decoction pieces. The result is expected to provide a reference for the establishment of quality standards for Zingiberis Rhizoma Carbonisata decoction pieces and Zingiberis Rhizoma decoction pieces.

[Modulation of gut microbiota during alleviation of antibiotic-associated diarrhea with Zingiberis Rhizoma].

This study was aimed to explore the effect of Zingiberis Rhizoma extract on rats with antibiotic-associated diarrhea(AAD), and reveal the modulation of gut microbiota during alleviation of AAD. AAD rat model was successfully established by exposing rats to appropriate antibiotic mixed solution. Peficon(70 mg·kg~(-1)·d~(-1)) was used as positive control, then rats were treated with 200 mg·kg~(-1)·d~(-1) and 400 mg·kg~(-1)·d~(-1) of Zingiberis Rhizoma extract for low and high dosage groups of Zingiberis Rhizoma extract, respectively. The weight changes of the rats were observed, and the degree of diarrhea were evaluated by fecal score, 120 min fecal weight and fecal water content. Colon tissues for histopathological examination were stained with hematoxylin and eosin(HE), and 16 S rRNA sequencing analysis of gut microbiota was performed. The results showed that compared with the model group, the degree of diarrhea, indicated by fecal water content, fecal score, and 120 min fecal weight of positive control group, Zingiberis Rhizoma low-dose group and Zingiberis Rhizoma high-dose group were significantly ameliorated. And the treatment of Zingiberis Rhizoma could significantly improve the pathological condition of colon tissue in AAD rats, especially the high dose of Zingiberis Rhizoma. In addition, 16 S rRNA sequencing analysis of gut microbiota showed that the diversity and abundance of gut microbiota were significantly improved and the reco-very of gut microbiota was accelerated after given high-dose of Zingiberis Rhizoma, while no significant changes of alterations were observed after given Pefikon. Of note, compared with the pefikon group, the abundance and diversity of gut microbiota in Zingi-beris Rhizoma high-dose group were significantly elevated. At the phylum level, the abundance of Firmicutes in AAD rats increased and the abundance of Proteobacteria was decreased after the Zingiberis Rhizoma intervention. At the genus level, the abundance of Bacillus spp., Lachnoclostridium and Escherichia coli-Shigella were decreased, and the abundance of Lactobacillus spp., Trichophyton spp., and Trichophyton spp., etc., were increased. While compared with the AAD model group, there was no significant difference of gut microbiota after given Peficon. The results showed that Zingiberis Rhizoma exerted beneficial health effects against AAD, and positively affected the microbial environment in the gut of rats with AAD.

Quality control of Zingiberis Rhizoma and its processed products by UHPLC-Q-TOF/MS-based non-targeted metabonomics combining with SIBDV method.

Zingiberis Rhizoma (ZR) is a homologous plant with pungent tastes and aromas, which has unique nutritional value and tremendous application potentiality. Zingiberis Rhizoma Praeparatum (ZRP) and Carbonised Ginger (CG) are processed products of ZR through different processing methods, and they are commonly used ingredients in food supplements. This study used ZR, ZRP and CG from different batches to further understand composition differences after processing. Additionally, we performed non-targeted metabolomics-based profiling of gingerols by ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) in combination with multivariate analysis and compounds identification. In which, we developed a comprehensive SWATH-IDA bi-directionally verified (SIBDV) method integrating the advantages of Sequential Windowed Acquisition of all Theoretical fragment ions (SWATHTM) and traditional information-dependent acquisition (IDA) mode for characterization of gingerols. Potential chemical markers were selected by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) of chemometrics methods. After that, the threshold variable importance in projection (VIP) value and P value were employed to screen the valuable MS features for discriminating ZR, ZRP and CG. In total, 59 gingerols in the different samples were structurally identified. Results allowed the selection of 33 gingerols, which are nominated as novel markers for materials authentication in ZR, ZRP and CG. The analysis of the study showed that the content of gingerols showed a downward trend after processing, but shogaols and gingerone compounds had an upward trend, resulting in differences in application and pharmacodynamic efficacy. These findings provide promising perspectives in the quality control of ZR, ZRP and CG, as well as for laying the foundation in food design and development.

Herbal Textual Research on Zingiberis Rhizoma Recens in Famous Classical Formulas and Its Quality Evaluation / 中国实验方剂学杂志

ObjectiveTo provide references for the selection of Zingiberis Rhizoma Recens on the research of famous classical formulas and the reasonable uses for medicines and foods through herbal textural research and quality analysis of Zingiberis Rhizoma Recens from main producing areas in China. MethodBy consulting the ancient and modern literature， the name， origin， producing areas， harvest time， processing methods of Zingiberis Rhizoma Recens were summarized. According to the 2020 edition of Chinese Pharmacopoeia， the contents of 6-gingerol， 8-gingerol， 10-gingerol， and volatile oil in Zingiberis Rhizoma Recens samples were determined. ResultHerbal textural research indicated that medicinal Zingiberis Rhizoma Recens originated from the fresh rhizome of Zingiber officinale. Before Tang dynasty， Zingiberis Rhizoma Recens produced in Sichuan was the best. In the Song dynasty， Zingiberis Rhizoma Recens produced in Sichuan， Zhejiang， and Anhui was of excellent quality. The cultivation of Zingiberis Rhizoma Recens in Shandong developed during the Ming and Qing dynasties. From ancient times to the present， the harvest period extended from the autumnal equinox to the winter solstice. Quality evaluation standards of Zingiberis Rhizoma Recens were essentially the same in ancient and present documents， as those with little gluten or gluten-free and strong pungency were preferred. After determination， the contents of 6-gingerol， 8-gingerol， and 10-gingerol in 44 samples were qualified in 27 samples， with a qualified rate of 61.4%. Among them， 17 samples were unqualified in the total contents of 8-gingerol and 10-gingerol. Among these qualified samples， the content of 6-gingerol ranged from 0.067% to 0.255%， and the total contents of 8-gingerol and 10-gingerol ranged from 0.040% to 0.131%. The content of volatile oil in 36 samples were qualified in 33 samples， with a qualified rate of 91.7%. Among the qualified samples， the content of volatile oil ranged from 0.175% to 0.410%. ConclusionZingiberis Rhizoma Recens has been used as medicines and foods since ancient times， and the genuine producing areas are consistent in ancient and present times， while the quality of the products， especially the medicinal Zingiberis Rhizoma Recens， should be monitored. Medicinal Zingiberis Rhizoma Recens planted in Leshan city of Sichuan province contains high contents of effective components， followed by Qujing and Wenshan cities of Yunnan province. Zingiberis Rhizoma Recens planted in Shandong and other places is mostly edible.

Serum Metabonomics Analysis of Diarrhea Mice Intervened by Zingiberis Rhizoma Recens Based on UPLC-QE-Orbitrap-MS / 中国实验方剂学杂志

ObjectiveTo investigate the antidiarrheal effect and mechanism of Zingiberis Rhizoma Recens on diarrhea mice， and to provide research basis for the inhibition of intestinal peristalsis by Zingiberis Rhizoma Recens and its application in the treatment of gastrointestinal diseases. MethodThe diarrhea model of mice was established by Sennae Folium. The control group， model group， Zingiberis Rhizoma Recens low-， medium-， high-dose groups （0.1， 0.32， 1.0 g·kg-1） and loperamide group （1.6 g·kg-1） were set. The intervention effect of Zingiberis Rhizoma Recens with different doses on diarrhea mice was detected by diarrhea score， incidence rate of loose stools （LSIR）， grade of average loose stools （ALSG）， diarrhea index （DI）， intestinal propulsion rate and intestinal pathological section. The serum metabonomics of mice was analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA）. The conditions were as follows：mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B）， flow rate of 0.4 mL·min-1， injection volume of 5 µL， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 100-1 500. ResultCompared with the model group， Zingiberis Rhizoma Recens high-dose group could obviously reduce the diarrhea score， LSIR， ALSG， DI and intestinal propulsion rate （P<0.05， P<0.01）， and improve the intestinal mucosal injury. There were 40 main differential metabolites among the control group， model group and Zingiberis Rhizoma Recens high-dose group， including glucose 1-phosphate， xanthine， xanthosine and so on. The metabolic pathways mainly included starch and sucrose metabolism， amino sugar and nucleotide sugar metabolism， fructose and mannose metabolism， tryptophan metabolism， and galactose metabolism. ConclusionZingiberis Rhizoma Recens can inhibit intestinal peristalsis in diarrhea mice and exert antidiarrhoea effect， the mechanism of which may be related to the regulation of carbohydrate and amino acid metabolism.

Effect of Asari Radix et Rhizoma-Zingiberis Rhizoma Herb Pair on Lung and Liver Lipid Metabolism in Rats with COPD Based on Metabolomics and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Asari Radix et Rhizoma-Zingiberis Rhizoma herb pair （XGHP） on lung and liver lipid metabolism in rats with chronic obstructive pulmonary disease （COPD）. MethodForty SD male rats were divided into a normal group （10 rats） and a model group （30 rats）. The method of cigarette smoke + tracheal injection of lipopolysaccharide（LPS） + cold stimulation was used to replicate COPD model with the syndrome of cold phlegm obstruction in lung. A COPD group， a XGHP group （5.4 g·kg-1·d-1）， and an aminophylline group （0.5 g·kg-1·d-1） were established after successfully inducing the model， with 10 rats in each group. After treatment， the serum triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） levels of rats in each group were measured. Gas chromatography-mass spectrometer （GC-MS） was used to detect the differential metabolites in the lung and liver tissues of rats in each group， and the relevant targets of the differential metabolites were predicted by network pharmacology. Molecular docking was used to verify the binding ability of key components in XGHP to the relevant targets in network pharmacology. The mRNA and protein expression levels of peroxisome proliferator-activated receptor α （PPARα） and fatty acid binding protein 4 （FABP4） in lung and liver tissues of rats in each group were detected by real-time polymerase chain reaction （PCR） and Western blot. ResultXGHP significantly increased the levels of TG， TC， and LDL-C in serum （P<0.05）， and decreased the level of HDL-C （P<0.05） in rats with COPD. GC-MS results showed that there were 8 lung differential metabolites and 17 liver differential metabolites in the COPD group and XGHP group. Network pharmacology predicted 59 common targets for the two differential metabolites， mainly enriched in the PPAR signaling pathway. Molecular docking results showed that the main components in XGHP were well combined with both PPARα and FABP4. Real-time PCR showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 mRNA （P<0.05）， and Western blot showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 proteins （P<0.05） in lung and liver tissues of rats with COPD. ConclusionXGHP effectively improves the blood lipid levels of rats with COPD， which may be related to the increase of the expression levels of PPARα and FABP4 mRNA and proteins in the PPAR signaling pathway， thus regulating lung and liver lipid metabolism.

Effect of Asari Radix et Rhizoma-Zingiberis Rhizoma Herb Pair on Lung and Liver Lipid Metabolism in Rats with COPD Based on Metabolomics and Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo investigate the effects of Asari Radix et Rhizoma-Zingiberis Rhizoma herb pair （XGHP） on lung and liver lipid metabolism in rats with chronic obstructive pulmonary disease （COPD）. MethodForty SD male rats were divided into a normal group （10 rats） and a model group （30 rats）. The method of cigarette smoke + tracheal injection of lipopolysaccharide（LPS） + cold stimulation was used to replicate COPD model with the syndrome of cold phlegm obstruction in lung. A COPD group， a XGHP group （5.4 g·kg-1·d-1）， and an aminophylline group （0.5 g·kg-1·d-1） were established after successfully inducing the model， with 10 rats in each group. After treatment， the serum triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） levels of rats in each group were measured. Gas chromatography-mass spectrometer （GC-MS） was used to detect the differential metabolites in the lung and liver tissues of rats in each group， and the relevant targets of the differential metabolites were predicted by network pharmacology. Molecular docking was used to verify the binding ability of key components in XGHP to the relevant targets in network pharmacology. The mRNA and protein expression levels of peroxisome proliferator-activated receptor α （PPARα） and fatty acid binding protein 4 （FABP4） in lung and liver tissues of rats in each group were detected by real-time polymerase chain reaction （PCR） and Western blot. ResultXGHP significantly increased the levels of TG， TC， and LDL-C in serum （P<0.05）， and decreased the level of HDL-C （P<0.05） in rats with COPD. GC-MS results showed that there were 8 lung differential metabolites and 17 liver differential metabolites in the COPD group and XGHP group. Network pharmacology predicted 59 common targets for the two differential metabolites， mainly enriched in the PPAR signaling pathway. Molecular docking results showed that the main components in XGHP were well combined with both PPARα and FABP4. Real-time PCR showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 mRNA （P<0.05）， and Western blot showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 proteins （P<0.05） in lung and liver tissues of rats with COPD. ConclusionXGHP effectively improves the blood lipid levels of rats with COPD， which may be related to the increase of the expression levels of PPARα and FABP4 mRNA and proteins in the PPAR signaling pathway， thus regulating lung and liver lipid metabolism.

Quality markers of Zingiberis Rhizoma Carbonisata before and after processing / 中国中药杂志

Based on the previous research results of our group and literature research, the chemical components, mechanisms, pharmacodynamics, and pharmacokinetics of Zingiberis Rhizoma Carbonisata were summarized to determine the quality markers(Q-markers) of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Our research group has clarified the differential components of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma, the meridian-warming hemostatic effect of Zingiberis Rhizoma Carbonisata, the related targets and pathways of the effect, the endogenous biomarkers of Zingiberis Rhizoma Carbonisata, and the hemodynamic processes of Zingiberis Rhizoma Carbonisata and Zingiberis Rhizoma. Moreover, based on high-performance liquid chromatography-diode array detector-electrospray ionization mass spectrometry(HPLC-DAD-ESIMS), a method for determining the content of Q-mar-kers was established. In conclusion, the study finally determined that gingerone, 6-shogaol, and diacetyl-6-gingerol were the Q-mar-kers of Zingiberis Rhizoma Carbonisata decoction pieces, and 6-gingerol, 8-gingerol, and 10-gingerol were Q-markers of Zingiberis Rhizoma decoction pieces. The result is expected to provide a reference for the establishment of quality standards for Zingiberis Rhizoma Carbonisata decoction pieces and Zingiberis Rhizoma decoction pieces.