Mass spectrometry-based metabolomics reveal Dendrobium huoshanense polysaccharide effects and potential mechanism of N-methyl-N'-nitro-N-nitrosoguanidine -induced damage in GES-1 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Dendrobium huoshanense C. Z. Tang et S. J. Cheng is an important edible medicinal plant that thickens the stomach and intestines, and its active ingredient, polysaccharide, can have anti-inflammatory, immunoregulatory, and antitumor effects. However, the gastroprotective effects and potential mechanisms of Dendrobium huoshanense polysaccharides (DHP) remain unclear. AIM OF THE STUDY: An N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) induced human gastric mucosal epithelial cells (GES-1) damage model was used in this research, aiming to investigate whether DHP has a protective effect on MNNG-induced GES-1 cell injury and its underlying mechanism based on the combination of multiple methods. MATERIALS AND METHODS: DHP was extracted using water extraction and alcohol precipitation methods, and the proteins were removed using the Sevag method. The morphology was observed using scanning electron microscopy. A MNNG-induced GES-1 cell damage model was developed. Cell viability and proliferation of the experimental cells were investigated using a cell counting kit-8 (CCK-8). Cell nuclear morphology was detected using the fluorescent dye Hoechst 33342. Cell scratch wounds and migration were detected using a Transwell chamber. The expression levels of apoptosis proteins (Bcl-2, Bax, Caspase-3) in the experimental cells were detected by Western blotting. Ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) was performed to investigate the potential mechanism of action of DHP. RESULTS: The CCK-8 kit analysis showed that DHP increased GES-1 cell viability and ameliorated GES-1 cell injury by MNNG. In addition, scratch assay and Transwell chambers results suggested that DHP improved the MNNG-induced motility and migration ability of GES-1 cells. Likewise, the results of the apoptotic protein assay indicated that DHP had a protective effect against gastric mucosal epithelial cell injury. To further investigate the potential mechanism of action of DHP, we analyzed the metabolite differences between GES-1 cells, GES-1 cells with MNNG-induced injury, and DHP + MMNG-treated cells using UHPLC-HRMS. The results indicated that DHP upregulated 1-methylnicotinamide, famotidine, N4-acetylsulfamethoxazole, acetyl-L-carnitine, choline and cer (d18:1/19:0) metabolites and significantly down-regulated 6-O-desmethyldonepezil, valet hamate, L-cystine, propoxur, and oleic acid. CONCLUSIONS: DHP may protect against gastric mucosal cell injury through nicotinamide and energy metabolism-related pathways. This research may provide a useful reference for further in-depth studies on the treatment of gastric cancer, precancerous lesions, and other gastric diseases.

Comparative analysis and chemical profiling of different forms of Peucedani Radix.

Peucedani Radix, derived from roots of Peucedanum praeruptorum Dunn, is a well-known herb used for centuries in traditional Chinese medicine, which is rich in various coumarins. Four different forms including "earthworm head", "bamboo-like", taproots and multi-branched roots have occurred in current producing areas, but the differences in their external features and chemistry have not been concerned till now. In this study, the morphological and microscopic characters of "earthworm head" and "bamboo-like" roots were compared in detail, and qualitative and quantitative characterization of main active coumarins in different forms of Peucedani Radix have been developed by UPCL-Q/TOF-MS and HPLC-DAD, respectively. The results showed that both "earthworm head" and "bamboo-like" consisted of rhizome and root, exhibiting distinct features from normal taproots. Moreover, 53 coumarins including simple coumarins, furanocoumarins and pyranocoumarins were identified or putatively characterized from the four forms samples under the established UPLC-Q/TOF-MS conditions. In addition, the developed quantitative method was successfully applied to simultaneously determine eight main coumarins in 24 batches of four forms Peucedani Radix and 12 batches of dissected "earthworm head" and "bamboo-like" samples. The quantitative results proved that wild "earthworm head" showed higher content of active coumarins and the "bamboo-like" was a rich source of coumarins in cultivated samples, especially for furanocoumarins. These findings would provide reasonable basis for further quality evaluation, grades classification and comprehensive utilization of P. praeruptorum resources.

[Planting area estimation of Paeonia ostii based on RS--by case study in Dangshan of Anhui province].

Moutan Cortex is one kind of famous medicinal materials. The dry root bark of Paeonia ostii which is a genuine medicinal material produced in Tongling,Anhui province,and later was introduced to Heze,Shandong province and Bozhou,Anhui province.Dangshan county is located at the northern end of Anhui province and adjacent to Shandong province. Its medicinal seedlings were came from Heze,Shandong province. At present,there is a lack of scientific investigation on the planting area of P. ostii in north China plain. On the basis of field investigation and remote sensing technology,through the data source provided by the remote sensing image of " Resources 3"( ZY-3),combined with the biological characteristics of P. ostii,the planting area of P. ostii in Dangshan county was extracted by field investigation and supervisory classification. The supervise classification method with the highest interpretation accuracy so far,the overall accuracy was 97. 81%,Kappa coefficient 0. 96. The results showed that the remote sensing classification method based on the maximum likelihood classification could extract P. ostii plots in the study area effectively. This study provides a scientific basis for the protection and rational utilization of traditional Chinese medicine resources,the development policy of traditional Chinese medicine industry and the long-term development plan in Dangshan county,and provides technical support for the poverty alleviation of traditional Chinese medicine industry in Dangshan county. It provides scientific reference for the application of remote sensing technology to investigate the planting area of P. ostii in in north China plain.

Determination of the species status of Fallopia multiflora, Fallopia multiflora var. angulata and Fallopia multiflora var. ciliinervis based on morphology, molecular phylogeny, and chemical analysis.

Relationships among Fallopia multiflora (Thunb.) Haraldson., F. multiflora var. angulata (S. Y. Liu) H. J. Yan, Z. J. Fang & Shi Xiao Yu., and F. multiflora var. ciliinervis (Nakai) Yonekura & H. Ohashi. were determined based on macroscopic and microscopic morphology, molecular phylogeny, and chemical analysis. The macroscopic and microscopic morphologies of root tubers or rhizomes, stems, and leaves were compared among the three taxa. The content of 11 chemical components (catechin, polydatin, stilbene glucoside, emodin, emodin-8-O-ß-D-glucopyranoside, rhein, chrysophanol, aloe-emodin, quercetin, physcion, and resveratrol) in the three taxa was determined by HPLC, and the chemical diversity was further evaluated by principal component and hierarchical cluster analyses. Molecular phylogenies were mapped using two chloroplast markers (matK and the psbA-trnH intergenic region) and a nuclear ribosomal marker [internal transcribed spacer 2 (ITS2) region]. Analyses of macroscopic and microscopic morphological characteristics revealed that the subterranean organs of F. multiflora and F. multiflora var. angulata are root tubers, whereas those of F. multiflora var. ciliinervis are rhizomes. In the phylogenetic trees, F. multiflora and F. multiflora var. angulata were clustered into a clade based on the combine matK + psbA-trnH sequence, with neighbour-joining, maximum likelihood, and Bayesian inference bootstrap support values of 99, 85, and 0.99, respectively. In addition, there were obvious differences in the chemical compositions of F. multiflora, F. multiflora var. angulata and F. multiflora var. ciliinervis. The root tubers of F. multiflora contain higher levels of stilbene glucoside and catechin, but lower levels of polydatin and anthraquinone compounds. In contrast to F. multiflora, the rhizomes of F. multiflora var. ciliinervis contain higher levels of polydatin and anthraquinone compounds, but lack stilbene glucoside. The content of all 11 assessed components was lower in F. multiflora var. angulata than in F. multiflora and F. multiflora var. cillinervis. Principal component and hierarchical cluster analyses revealed that F. multiflora and F. multiflora var. angulata individuals were clustered into a single clade, whereas F. multiflora var. ciliinervis individuals were clustered into a single clade separate from that containing F. multiflora and F. multiflora var. angulata individuals. On the basis of the results of our morphological, molecular phylogeny, and chemical analyses, we tentatively conclude that F. multiflora var. ciliinervis is an independent species, whereas F. multiflora var. angulata should be considered as a variety of F. multiflora.