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.

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.

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

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.

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.

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.

Modulation of gut microbiota during alleviation of antibiotic-associated diarrhea with Zingiberis Rhizoma / 中国中药杂志

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 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.

A new monoterpene ester from Zingiberis Rhizoma Recens / 中国中药杂志

Five monoterpenoid compounds(1-5) were isolated and purified from the acetone fraction of the aqueous extract of Zingiberis Rhizoma Recens by MCI, Sephadex LH-20, silica gel, semi-preparative HPLC, and TLC. Their structures were identified with multiple spectroscopical methods including 1 D-NMR, 2 D-NMR, and MS. The five compounds were identified as(2E,6Z)-8-hydroxy-2,6-dimethylocta-2,6-dien-1-yl-(E)-3-(4-hydroxy-3-methoxyphenyl) acrylate(1),(2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-die-noic acid(2),(E)-1,8-dihydroxy-3,7-dimethyl-2-octenoic acid(3), linalyl-β-D-glucopyranoside(4), and β-D-glucopyranoside-(2E)-3,7-dimethyl-2,6-octadien-1-yl(5), respectively.Compound 1 was a new monoterpene ester, and compounds 4-5 were isolated from this plant for the first time.

Effect of Coptidis Rhizoma-Zingiberis Rhizoma Extract on Colonic Epithelium TLR4/NF-κB Signaling Pathway in Mice with Ulcerative Colitis / 中国实验方剂学杂志

Objective::To observe the effect of berberine and 6-shogaol, main components of Coptiae Rhizoma and Zingiberis Rhizoma, on the inflammatory signaling pathway of Toll-like receptors 4 (TLR4)/nuclear factor kappa B (NF-κB) in colonic epithelial cells of mice with ulcerative colitis. Method::Fifty Kunming mice were randomly divided into normal group, model group, berberine group (100 mg·kg-1), 6-shogaol group (100 mg·kg-1), and 6-shogaol combined with berberine group (200 mg·kg-1), with 10 mice in each group. A mouse model of ulcerative colitis was established through oral administration with 2% dextroan sulfate for two weeks. Each group was given corresponding drugs by gavage, while normal group and model group were given equal amount of normal saline. Serum and colon tissue samples were taken 20 days after administration. Enzyme-linked immunosorbent method was used to detect serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) expressions, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot method were used to detect TLR4, NF-κB p65 mRNA and protein expressions in colon epithelial tissue. Result::Compared with the normal group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were increased in the model group (P<0.01), and the contents of serum IL-1β and TNF-α were increased (P<0.01). Compared with the model group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were significantly decreased in 6-shogaol group, berberine group and 6-shogaol combined with berberine group (P<0.01), and the contents of serum IL-1β and TNF-α were significantly decreased (P<0.01). Among the three groups, 6-shogaol combined with berberine group had the strongest effect (P<0.01). Conclusion::Both 6-shogaol and berberine can inhibit colonic inflammation, reduce inflammatory damage and treat ulcerative colitis. The combined application of 6-shogaol and berberine has a significant synergism effect. The mechanism is related to the excessive activation of TLR4/NF-κB pathway and the regulation of non-controllable intestinal inflammation.

Five new gingerol compounds from Zingiberis Rhizoma / 药学学报

We separated and purified five chemical constituents of dried ginger by Diaion HP-20, Sephadex LH-20, silica gel and semi-preparative high performance liquid chromatography. Five gingerols were identified by physicochemical properties and MS and NMR spectroscopy techniques: 4-(2-butyl-6-methyl-4H-pyran-4-yl)-2-methoxyphenol (1), 4-(2-hexyl-6-methyl-4H-pyran-4-yl)-2-methoxyphenol (2), 1-(4-hydroxy-3-methoxyphenyl)tridecane-3,5-diol (3), [10]-gingerdiol (4) and 1-[1-(4-hydroxy-3-methoxy phenyl)-3-oxodecan-5-yl]pyrrolidin-2-one (5a, 5b). Compounds 1-3, 5a, 5b are new compounds.

Study on the attenuation mechanism of aristolochia manshuriensis compatibility with zingiberis rhizoma based on different methods of boiling and blending / 国际中医中药杂志

Objective@#Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed.@*Methods@#The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method.@*Results@#There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ± 0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01).@*Conclusions@#The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.

Study on the attenuation mechanism of aristolochia manshuriensis compatibility with zingiberis rhizoma based on different methods of boiling and blending / 国际中医中药杂志

Objective Comparison of effects of Zingiberis Rhizoma and Aristolochia manshuriensis on diuretic effect and acute renal injury in rats by combining two methods of co-decoction and mixed-decoction. The effects of in vitro observation on normal human renal tubular epithelial cells (HK-2) were observed. Methods The rats were randomly divided into five groups: blank group, positive control group, aristolochia manshuriensis group, combined decoction group and divided decoction group, 8 rats in each group. The water-loading rat model was established by intragastric administration of normal saline. The urine of rats was collected and the volumes of urine were measured for 24 hours after the corresponding drugs were given to each group. After 2 weeks of gavage of the corresponding drugs in each group, the serum BUN, SCr and urine UCr and PRO levels were measured by 7600P automatic biochemical analyzer, and renal histopathology were observed by HE staining. The HK-2 cells were cultured in vitro and divided into control group, Aristolochia manshuriensis group, mixed decoction group and sub-decoction group. After 24 hour intervention, the activity of cells was detected by CCK-8 method and the apoptosis was observed by Hoechst stain method. Results There was no significant difference in 24 hour urine output between the groups (P>0.05). Compared with Aristolochia manshuriensis group, the kidney coefficient (0.010 1 ±0.005 8 vs. 0.013 3 ± 0.007 8), SCr (38.52 ± 0.58 μmol/L vs. 46.61 ± 0.72 μmol/L), BUN (8.55 ± 0.12 mmol/L vs. 10.21 ± 0.30 mmol/L), UCr (52.21 ± 0.89 μmol/L vs. 57.71 ± 0.67 μmol/L), PRO (29.89 ± 0.18 mg/L vs. 34.23 ± 6.05 mg/L) of combined decoction group significantly decreased (P<0.05). The survival rate of HK-2 cells (72.45% ± 3.70% vs. 55.92% ± 8.39%) in combined decoction group significantly increased (P<0.01), and the apoptosis rate (7.9% ± 2.6% vs. 31.6% ± 9.1%) significantly decreased (P<0.01). Conclusions The traditional co-decoction method of Aristolochia manshuriensis compatibility with Zingiberis Rhizoma can achieve a certain attenuation effect, and the mixed-decoction group can not achieve the attenuating effec.

Study on a high-throughput method with LC-MS/MS technique for detecting aflatoxins in ginger / 国际中医中药杂志

Objective@#To optimize the method of simultaneous determination of four aflatoxins (B1, B2, G1 and G2) of ginger by the ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method and high-throughput method.@*Methods@#The aflatoxins were extracted from ginger by methanol-water (80:20, V/V) solution, concentrated and dried with nitrogen. The aflatoxins were detected by UPLC-MS/MS by using Waters Acquity UPLC BEH C18 chromatographic column. The mobile phase was 0.1% formic acid water (A phase) -0.1% formic acid methanol (B phase), gradient elution, flow rate 0.35 ml/min, mass spectrometry was electrospray ion source, positive ion scanning mode, multi reaction ion monitoring were using.@*Results@#Quantification of four aflatoxins by matrix matching standard curve. The linear was good in the range of 0.125-20.000 ng/ml, and the correlation coefficients were all greater than 0.999 0. The ginger sample detection was 0.125-0.300 μg/kg and 0.125-1.000 μg/kg, respectively. The average recoveries were 81.7%-96.0%, and the relative standard deviation (RSD) was lower than 7.53%.@*Conclusions@#This method is simple, rapid, sensitive and low limit of detection, which can meet the requirements for the detection of trace aflatoxins residues in ginger.

Quality Evaluation of Standard Decoction of Zingiberis Rhizoma / 中国实验方剂学杂志

Objective: To establish the quality control methods for the standard decoction of Zingiberis Rhizoma. Method: DNA barcode primitives were identified for the medicinal materials from different origins; according to the standard of Chinese herbal medicine decoction preparation principle,the identified Zingiberis Rhizoma was prepared into standard decoction for analysis. Meanwhile, the extraction method and analysis method were validated from methodologies, and the transfer rate of 6-gingerol as well as the extraction rate of standard decoction of Zingiberis Rhizoma were calculated. In addition,the quality standard of standard decoction of Zingiberis Rhizoma was also established based. The structures of main chromatographic peaks were identified by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to clarify the main chemical constituents in the standard decoction of Zingiberis Rhizoma. Result: All the samples were identified as Zingiberis Rhizoma. Under the conditions established in this paper,the standard curve of 6-gingerol was Y=661.56X+2.493 3(r=0.999 3),and the RSD was 0.5%in precision test, indicating that the instrument precision was good. The repeatability test showed that the RSD was 0.3%, indicating that the method had good repeatability. The stability test showed that the RSD was 0.4%, indicating that the test solution had good stability within 24 h. The recovery rate was 97.2%and the RSD was 0.6%,indicating that the method was accurate and reliable. 6-gingerol's transfer rate ranged from 31.8%to 57.4%and the extraction rate was within the range of 9.6%-23.1%. The fingerprint similarity of 12 batches of Zingiberis Rhizoma standard decoction was>90%. Conclusion: The established quality control method for Zingiberis Rhizoma was stable and feasible; meanwhile, the standard preparation method for Zingiberis Rhizoma and its quality evaluation system were also established in this study.

Effect of Zingiberis Rhizoma Recens on Hemorrhoids Model in Rats and Mice / 中国实验方剂学杂志

Objective: To explore the effect of Zingiberis Rhizoma Recens(ZRR) on hemorrhoids in mice and rats. Method: Sixty SPF-grade SD rats were divided into blank group, model group, Ma Yinglong Shexiang hemorrhoid ointment group (7.5 g·kg-1), and large and small-dose ZRR paste groups (10, 5 g·kg-1). ZRR paste was applied in Yongquan acupoint to observe the effect of 0.05 mL injection with 75% acetic acid solution on hemorrhoids induced by subcutaneous anus in rats, the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), nitric oxide (NO) in the serum were detected by enzyme linked immunosorbent assay(ELISA), and the rectal histopathology was detected by hematoxylin-eosin(HE). Sixty SPF-grade KM mice were divided into blank group, model group, MA Ying-long Shexiang hemorrhoid ointment group (7.5 g·kg-1), and large and small-dose ginger essential oil groups (0.06, 0.03 mL, three times a day). ZRR paste was applied in crissum to observe the effect of injection of 20% acetic acid solution 0.05 mL (maintaining for 1 min) on hemorrhoids in mice induced by anus. The degree of local swelling ulcer around the anus and score was observed, levels of IL-1β, TNF-α in the serum were determined by ELISA, and the rectal histopathology was detected by HE staining. Result: In the experimental study on treating hemorrhoids with ZRR paste applied on Yongquan point of rats, compared with normal group, serum levels of IL-1β, IL-6, TNF-α, NO in model group were significantly higher (Pβ, IL-6, TNF-α, NO were decreased in each administration group (PPPPβ, TNF-α in model group were significantly higher (PPβ, TNF-α levels (PPConclusion: External application of ZRR can effectively inhibit perianal swelling and ulcer degree, with a good therapeutic effect on hemorrhoids model in rats and mice.

High-throughput screening of ochratoxin A in Chinese herbal medicines using enzyme-linked immunoassay / 中国中药杂志

An indirect competitive enzyme-linked immunosorbent assay( ic-ELISA) was developed for the rapid detection of ochratoxin A( OTA) in nutmeg( Myristicae Semen),ginger( Zingiberis Rhizoma) and turmeric( Curcumae Longae Rhizoma). The matrix matching standard curve was used instead of the standard curve of sample diluent,and the sample extract and sample diluent were optimized. The sensitivity( IC_(50)) of this method for OTA in nutmeg,ginger and turmeric were determined as 0. 146,0. 157 and 0. 153 ng·m L~(-1),respectively and the limits of detection( LODs) were 0. 040,0. 032 and 0. 031 ng·m L~(-1),respectively. The recovery of samples ranged from 75. 99% to 122. 3%,with RSD<10%. Two positive samples for nutmeg and one positive sample for turmeric occurred in 50 samples,and the highest OTA contamination value was 1 167. 8 μg·kg~(-1). The results were further confirmed by LC-MS/MS. It shows that the developed ic-ELISA method is simple,rapid and sensitive,and can be applied for rapid and high-throughput screening of OTA in nutmeg,ginger and turmeric,as well as some other CHMs.

Isolation and Identification of Gingerols Constituents from Zingiber officinale Rosc / 中国药学杂志

OBJECTIVE: To study the chemical constituents of Zingiber officinale Rosc. METHODS: Various column chromatographic techniques were used to isolate and purify the chemical constituents and their structure were elucidated by spectral analysis. RESULTS: Ten compounds were isolated and identified as 6-shogoal (I), 9-shogoal (II), 10-shogoal (III), 12-paradol (IV), 6-gingerol (V), 6-isodehydrogingerdione (VI), 8-isodehydrogingerdione (VII), 5-hydroxy-1-(4'-hydroxy-3'-methoxyphenyl)-4-hexa-decen-3-one (VIII), (E)-geranylferulic acid (IX), and 5-ethoxy-1-(4-hydroxy-3-methoxyphenyl)tetradecan-3-one (X). CONCLUSION: The compounds II, III, IV, VI, VII and X are isolated from Zingiberis Rhizoma for the first time.

Study on Chemical Constituents from Zingiberis Rhizoma / 中国药房

OBJECTIVE:To study the chemical constituents of Zingiberis Rhizoma.METHODS:The compounds of Zingiberis Rhizoma were classified and purified by silica gel,Sephadex LH-20 column chromatography and thin layer chromatography (TLC).The structure of compounds were analyzed and identified according to chemical property and spectrum data.RESULTS & CONCLUSIONS:Five compounds were isolated from Zingiberis Rhizoma,i.e.methyl-6-gingerol (1),4-gingerol (2),β-eudesmol (3),2,5-dihydroxybisabola-3,10-diene (4),6-shogaol (5).The compounds 2,3,4 are isolated from Zingiberis Rhizoma for the first time.