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.

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.

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.

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.

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.

Extraction of ginger oleoresin from Zingiberis Rhizoma Recens and inclusion of ginger oleoresin with β-cyclodextrin / 中国药学杂志

OBJECTIVE: To optimize the extraction and inclusion processes of ginger oleoresin from Zingiberis Rhizoma Recens. METHODS: Supercritical carbon dioxide extraction was selected to extract ginger oleoresin from Zingiberis Rhizoma Recens. The extraction condition was optimized by orthogonal experiment. The inclusion method, the ratio of ginger oleoresin to β-cyclodextrin and inclusion time were studied as factors of inclusion process. RESULTS: The optimized extraction technology was as follows: extraction temperature was 50℃, extraction pressure was 25 MPa, separation pressure was 9 MPa and extraction time was 90 min. The optimal inclusion method was triturating inclusion, with the ratio of ginger oleoresin to β-cyclodextrin of 1:8 and inclusion time of 60 min. CONCLUSION: The optimized technology of extraction and inclusion is stable and feasible, and can be used for the extraction and inclusion of ginger oleoresin.