Neuroprotective Effects and Metabolomics Study of Protopanaxatriol (PPT) on Cerebral Ischemia/Reperfusion Injury In Vitro and In Vivo.

Stroke, one of the leading causes of disability and death worldwide, is a severe neurological disease that threatens human life. Protopanaxatriol (PPT), panaxatriol-type saponin aglycone, is a rare saponin that exists in Panax ginseng and Panax Noto-ginseng. In this study, we established an oxygen-glucose deprivation (OGD)-PC12 cell model and middle cerebral artery occlusion/reperfusion (MCAO/R) model to evaluate the neuroprotective effects of PPT in vitro and in vivo. In addition, metabolomics analysis was performed on rat plasma and brain tissue samples to find relevant biomarkers and metabolic pathways. The results showed that PPT could significantly regulate the levels of LDH, MDA, SOD, TNF-α and IL-6 factors in OGD-PC12 cells in vitro. PPT can reduce the neurological deficit score and infarct volume of brain tissue in rats, restore the integrity of the blood-brain barrier, reduce pathological damage, and regulate TNF-α, IL-1ß, IL-6, MDA, and SOD factors. In addition, the results of metabolomics found that PPT can regulate 19 biomarkers involving five metabolic pathways, including amino acid metabolism, arachidonic acid metabolism, sphingolipid metabolism, and glycerophospholipid metabolism. Thus, it could be inferred that PPT might serve as a novel natural agent for MCAO/R treatment.

Protective Effects of Protopanaxatriol Saponins on Ulcerative Colitis in Mouse Based on UPLC-Q/TOF-MS Serum and Colon Metabolomics.

Ulcerative colitis (UC) is a chronic, nonspecific inflammation of the bowel that mainly affects the mucosa and submucosa of the rectum and colon. Ginsenosides are the main active ingredients in ginseng and show many therapeutic effects in anti-inflammatory diseases, cancer, and nervous system regulation. Protopanaxatriol saponin (PTS) is an important part of saponins, and there is no research on its pharmacological effects on colitis. In this study, a model of ulcerative colitis in mice was induced by having mice freely drink 3.5% dextran sodium sulfate (DSS) solution, and UPLC-Q-TOF-MS-based metabolomics methods were applied to explore the therapeutic effect and protective mechanism of PTS for treating UC. The results showed that PTS could significantly prevent colon shortening and pathological damage and alleviate abnormal changes in UC mouse physiological and biochemical parameters. Moreover, PTS intervention regulated proinflammatory cytokines such as TNF-α, IL-6, and IL-1 in serum, and MPO and NO in colon. Interestingly, PTS could significantly inhibit UC mouse metabolic dysfunction by reversing abnormal changes in 29 metabolites and regulating eleven metabolic pathways. PTS has potential application in the treatment of UC and could alleviate UC in mice by affecting riboflavin metabolism, arachidonic acid metabolism, glycerophospholipid metabolism, retinol metabolism, and steroid hormone biosynthesis and by regulating pentose and glucuronate conversion, linoleic acid metabolism, phenylalanine metabolism, ether lipid metabolism, sphingolipid metabolism, and tyrosine metabolism, which points at a direction for further research and for the development of PTS as a novel natural agent.

Comparative LC-LTQ-MS-MS Analysis of the Leaf Extracts of Lantana camara and Lantana montevidensis Growing in Egypt with Insights into Their Antioxidant, Anti-Inflammatory, and Cytotoxic Activities.

Lantana camara L. and Lantana montevidensis Briq. (F. Verbenaceae) are invasive ornamental weeds native to the tropical regions of Africa and America. The leaves of both species have been traditionally used as infusions for treating fever, rheumatism, and cancer. LC-MS-MS-guided profiling of the methanolic extracts of the leaves of L. camara and L. montevidensis growing in Egypt led to the putative identification of 59 compounds belonging to terpenoids, flavonoids, iridoid glycosides, phenolic acids, and their derivatives. The in-vitro antioxidants and anti-inflammatory and anticancer activities of the two extracts were investigated. L. camara and L. montevidensis inhibited DPPH• (IC50 = 34.01 ± 1.32 and 47.43 ± 1.74 µg/mL), ABTS+ (IC50 = 30.73 ± 1.42 and 40.37 ± 1.51 µg/mL), and superoxide anion (IC50 = 1.57 ± 0.19 and 1.31 ± 0.14 µg/mL) free radicals. A potent anti-inflammatory effect was observed for both species through the inhibition of elastase release in fMLF/CB-induced human neutrophils (IC50 = 2.40 ± 0.16 and 1.90 ± 0.07 µg/mL). The extracts showed significant cytotoxic activity against a panel of cancer cell lines with the most potent activity against Caco cells (IC50 = 45.65 ± 1.64 and 40.67 ± 1.52 µg/mL for L. camara and L. montevidensis, respectively). Western blotting supported by FACS analysis revealed that the extracts inhibited cancer cell proliferation, reduced metastasis, and induced apoptosis resulting in cell cycle arrest. This was achieved via increasing mRNA and protein expressions of p53 and GSK-3ß as well as decreasing the expression of PI3K, Akt, and cyclin D1.

Protective Effect of Ethyl Rosmarinate against Ulcerative Colitis in Mice Based on Untargeted Metabolomics.

Aiming at assessing the therapeutic effect of ethyl rosmarinate (ER) on ulcerative colitis (UC), the following activities were performed in vitro and in vivo in the present study. Firstly, a lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model was established to determine the level of inflammatory factors. Then, a UC mice model induced by dextran sodium sulfate (DSS) was established to further investigate the effects of ER on symptoms, inflammatory factors and colon histopathology. Finally, serum and colon metabolomics studies were performed to identify the biomarkers and metabolisms closely related to the protective effect of ER on UC. The results showed that after ER intervention, the levels of inflammatory factors (NO, TNF-α, IL-1ß and IL-6) and key enzyme (MPO) in cell supernatant, serum or colon were significantly decreased, and the disease activity index and colon tissue damage in mice were also effectively improved or restored. In addition, 28 biomarkers and 6 metabolisms were found to be re-regulated by ER in the UC model mice. Therefore, it could be concluded that ER could effectively ameliorate the progression of UC and could be used as a new natural agent for the treatment of UC.

Comprehensive phytochemicals analysis and anti-myocardial ischemia activity of total saponins of American ginseng berry.

American ginseng berry (AGB) is a new medicinal source. Total saponins of American ginseng berry (TSAGB) are the main active ingredients. The effects and active saponins of TSAGB on myocardial ischemia (MI) rats were evaluated for the first time. First, there were 69 saponins identified or tentatively characterized by Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS/MS) combined with UNIFI platform, among which, about 28 saponins were first identified in AGB. Second, MI model was established by ligating left coronary artery. It has been demonstrated that TSAGB could prevent the ST-segment elevation, reduce myocardial infarct size and levels of aspartate aminotransferase (AST), creatine kinase (CK), lactate dehydrogenase (LDH), malondialdehyde (MDA), and elevate the superoxide dismutase (SOD) level. Finally, network pharmacology combined with molecular docking to screen out four active saponins (ginsenoside Re, Rb3 , Rg3 , and PF11 ) and five key targets (SOD1, LDHA, CKB, GOT2, and ROS1) closely related to MI. PRACTICAL APPLICATIONS: This study enriches the chemical composition of TSAGB, and provides a basis for clarifying the pharmacological substances for anti-myocardial ischemia. TSAGB might be a potential anti-myocardial ischemia agent. The effect might be related to alleviating oxidative stress.

A new alkynol compound from Platycodins folium and its cytotoxicity.

A new alkynol, platycodynol (1), identified as 2, 3, 9, 13, 14-pentahydroxy-4, 6-tetradecadiyne, along with six known compounds (2-7) were obtained from Platycodins folium for the first time. Their structures were elucidated with infrared (IR), ultraviolet (UV), 1D and 2D nuclear magnetic resonance (NMR) spectroscopic analysis as well as by high resolution electrospray ionization mass spectroscopy (HRESIMS). Platycodynol showed cytotoxicity against S180, A549 and SPC-A-1 cancer cells but no cytotoxicity against normal cells NCTC1469 and HL-7702 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method.

Comprehensive Investigation on Metabolites of Wild-Simulated American Ginseng Root Based on Ultra-High-Performance Liquid Chromatography-Quadrupole Time-of-Flight Mass Spectrometry.

Aiming to evaluate the similarities and differences of the phytochemicals in different morphological regions of wild-simulated American ginseng (WsAG) root, the comprehensive metabolite profiling of main root (MR), branch root (BR), rhizome (RH), adventitious root (AR), and fibrous root (FR) was performed on the basis of ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry for the first time. First, in the screening analysis, a total of 128 shared compounds were identified or tentatively characterized. The results showed that these five parts were all rich in phytochemicals and contained similar structure types. Second, in the untargeted metabolomic study, it was found that there indeed existed differences between the MR&BR group, RH&AR group, and FR part when considering the contents of every ingredient. A total of 31 (12, 7, and 12 for MR&BR, RH&AR, and FR, respectively) potential chemical markers enabling the differentiation were discovered. This comprehensive phytochemical profile study revealed the structural diversity of secondary metabolites and the similar/different patterns in five morphological regions of WsAG root. It could provide chemical evidence for the rational application of different parts of WsAG root.

Non-Targeted Metabolomic Analysis of Methanolic Extracts of Wild-Simulated and Field-Grown American Ginseng.

Aiming at revealing the structural diversity of secondary metabolites and the different patterns in wild-simulated American ginseng (WsAG) and field-grown American ginseng (FgAG), a comprehensive and unique phytochemical profile study was carried out. In the screening analysis, a total of 121 shared compounds were characterized in FgAG and WsAG, respectively. The results showed that both of these two kinds of American ginseng were rich in natural components, and were similar in terms of the kinds of compound they contained. Furthermore, in non-targeted metabolomic analysis, when taking the contents of the constituents into account, it was found that there indeed existed quite a difference between FgAG and WsAG, and 22 robust known biomarkers enabling the differentiation were discovered. For WsAG, there were 12 potential biomarkers including two ocotillol-type saponins, two steroids, six damarane-type saponins, one oleanane-type saponins and one other compound. On the other hand, for FgAG, there were 10 potential biomarkers including two organic acids, six damarane-type saponins, one oleanane-type saponin, and one ursane. In a word, this study illustrated the similarities and differences between FgAG and WsAG, and provides a basis for explaining the effect of different growth environments on secondary metabolites.

UPLC-QTOF/MS-Based Nontargeted Metabolomic Analysis of Mountain- and Garden-Cultivated Ginseng of Different Ages in Northeast China.

Aiming at further systematically comparing the similarities and differences of the chemical components in ginseng of different ages, especially comparing the younger or the older and mountain-cultivated ginseng (MCG), 4, 5, 6-year-old cultivated ginseng (CG) and 12, 20-year-old MCG were chosen as the analytical samples in the present study. The combination of UPLC-QTOF-MSE, UNIFI platform and multivariate statistical analysis were developed to profile CGs and MCGs. By the screening analysis based on UNIFI, 126 chemical components with various structural types were characterized or tentatively identified from all the CG and MCG samples for the first time. The results showed that all the CG and MCG samples had the similar chemical composition, but there were significant differences in the contents of markers. By the metabolomic analysis based on multivariate statistical analysis, it was shown that CG4⁻6 years, MCG12 years and MCG20 years samples were obviously divided into three different groups, and a total of 17 potential age-dependent markers enabling differentiation among the three groups of samples were discovered. For differentiation from other two kinds of samples, there were four robust makers such as α-linolenic acid, 9-octadecenoic acid, linoleic acid and panaxydol for CG4⁻6 years, five robust makers including ginsenoside Re1, -Re2, -Rs1, malonylginsenoside Rb2 and isomer of malonylginsenoside Rb1 for MCG20 years, and two robust makers, 24-hydroxyoleanolic acid and palmitoleic acid, for MCG12 years were discovered, respectively. The proposed approach could be applied to directly distinguish MCG root ages, which is an important criterion for evaluating the quality of MCG. The results will provide the data for the further study on the chemical constituents of MCG.

[Absorbed components analysis of Zhitong Huazheng Capsules in rat serum by UPLC-Q-TOF/MS].

To identify and analyze the constituents in rat serum after oral administration of Zhitong Huazheng capsule (ZTHZC), and provide a reference for its further research on pharmacodynamics material basis. Female Wistar rats were selected as experimental animals, and received intragastric administration of ZTHZC at a dose of 1.5 g·kg⁻¹. After the serum samples were collected, the absorbed prototype components in rat serum were identified and analyzed by using ultra-high performance liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UPLC-Q-TOF/MS) combined with multivariate statistical analysis．The results showed, a total of fifteen absorbed constituents were identified, all of which were prototype components, including Danshensu, salvianolic acid A, B, C, D, 9,12-dihydroxy-15-nonadecanoicacid, linoleic acid, ethyl palmitoleate, tetrahydropalmatine, fumarate A, astragaloside A, astragaloside II, saponin, locustin and luteolin. This experiment showed that these fifteen components absorbed into blood may be the potential bioactive components in ZTHZC, providing a scientific basis for clarifying its material basis in pharmacodynamics.

[Determination of Heavy Metal Baseline Values and Analysis of Its Accumulation Characteristics in Agricultural Land in Chongqing].

In order to understand the accumulation of heavy metals of agricultural land in Chongqing and make clear priority control elements, the soil heavy metal baseline values need to be determined. Based on geochemical baselines, a total of 214 surface soil samples of six soil types were collected according to stratified sampling. Under strict quality control, the As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn content in each sample was measured. The baseline values of eight heavy metals were determined using mathematical statistics, iterative culling, and cumulative frequency curve methods. The accumulation of heavy metals in agricultural soils in Chongqing was evaluated using the geochemical baseline pollution and geological accumulation index methods. The mean baseline values of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn in agricultural land in Chongqing were 5.83, 0.25, 66.78, 25.45, 0.069, 29.90, 26.18, and 78.44 mg·kg-1, respectively, which agreed with the historical findings. Evaluation of the geochemical baseline pollution index method showed that the accumulation of As, Hg, and Zn in soil was the most prominent, with 14.65%, 11.82%, and 3.88% of the samples reaching severe accumulation, respectively. The proportions of moderate accumulation of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn were 12.12%, 6.50%, 3.79%, 6.50%, 15.27%, 5.29%, 5.45%, and 17.96%, respectively, of which As, Hg, and Zn were relatively large. The geological accumulation index indicated that more than 93% of the samples of Cd, Cr, Cu, Ni, and Pb were in a state of no accumulation. The proportions of mild accumulation of Hg, Zn, and As were 26.60%, 21.84%, and 21.21%, respectively. Only 5.56% and 0.49% of the samples had moderate accumulation of As and Zn. Different soil types have different properties. All soil types in the sample had different degrees of accumulation of the eight kinds of heavy metals. The accumulation of heavy metals in paddy and purple soils was lower than that in other soil types. For the soils of different geological and geomorphic types, the Ki values of As in the limestone middle and low mountain area of Daba Mountain in Northeast Chongqing and the low mountain area of the limestone of Wuling Mountain in Southeast Chongqing were relatively large; the rest were not significantly different, and all belonged to the non-accumulation and mild accumulation ranges. Both evaluation methods showed that the main accumulated heavy metals in agricultural land soil in Chongqing was As, followed by Hg and Zn, and management and prevention of the accumulation of these elements should be strengthened.

MicroRNA-139-5p inhibits cell proliferation and invasion by targeting insulin-like growth factor 1 receptor in human non-small cell lung cancer.

INTRODUCTION: Increasing evidence suggested that microRNAs (miRNAs) play a critical role in tumorigenesis. Decreased expression of miRNA-139-5p has been observed in various types of cancers. However, the biological function of miRNA-139-5p in non-small cell lung cancer (NSCLC) is still largely unknown. METHODS: Quantitative real-time PCR (qRT-PCR) was used to explore the expression level of miRNA-139-5p in NSCLC tissues and cell lines. Then, we investigated the role of miRNA-139-5p to determine its potential roles on lung cancer cell proliferation, migration and invasion in vitro. A luciferase reporter assay was performed to confirm the target gene of miRNA-139-5p and the results were validated in renal cancer cells. RESULTS: miRNA-139-5p was significantly decreased in NSCLC tissues and cell lines. Over-expression of miRNA-139-5p could inhibit lung cancer cell proliferation, migration, and invasion in vitro. Furthermore, we identified insulin-like growth factor 1 receptor (IGF1R) as a target of miR-139-5p and miR-139-5p function as a tumor suppressor via targeting IGF1R in NSCLC. CONCLUSIONS: Our results indicated that miR-139-5p acts as a tumor suppressor in NSCLC partially via down-regulating IGF1R expression.