Di Cao

New clerodane diterpenoids from Croton crassifolius.

Two new clerodane diterpenoids (1-2), one new clerodane diterpenoid alkaloid (3), as well as thirteen known compounds were isolated from Croton crassifolius. The structures of new compounds were established by a combination of spectroscopic methods, including HRMS, (1)H NMR, (13)C NMR, (1)H (1)H COSY, HSQC, HMBC, NOESY and X-ray crystallographic analysis. Compound 3 is firstly reported as the clerodane-type diterpenoid alkaloid in natural products. All of the compounds were evaluated for in vitro cytotoxic activities against CT26.WT cell using the MTT method.

Valproate induced hepatic steatosis by enhanced fatty acid uptake and triglyceride synthesis

ABSTRACT Steatosis is the characteristic type of VPA‐induced hepatotoxicity and may result in life‐threatening hepatic lesion. Approximately 61% of patients treated with VPA have been diagnosed with hepatic steatosis through ultrasound examination. However, the mechanisms underlying VPA‐induced intracellular fat accumulation are not yet fully understood. Here we demonstrated the involvement of fatty acid uptake and lipogenesis in VPA‐induced hepatic steatosis in vitro and in vivo by using quantitative real‐time PCR (qRT‐PCR) analysis, western blotting analysis, fatty acid uptake assays, Nile Red staining assays, and Oil Red O staining assays. Specifically, we found that the expression of cluster of differentiation 36 (CD36), an important fatty acid transport, and diacylglycerol acyltransferase 2 (DGAT2) were significantly up‐regulated in HepG2 cells and livers of C57B/6J mice after treatment with VPA. Furthermore, VPA treatment remarkably enhanced the efficiency of fatty acid uptake mediated by CD36, while this effect was abolished by the interference with CD36‐specific siRNA. Also, VPA treatment significantly increased DGAT2 expression as a result of the inhibition of mitogen‐activated protein kinase kinase (MEK) – extracellular regulated kinase (ERK) pathway; however, DGAT2 knockdown significantly alleviated VPA‐induced intracellular lipid accumulation. Additionally, we also found that sterol regulatory element binding protein‐1c (SREBP‐1c)‐mediated fatty acid synthesis may be not involved in VPA‐induced hepatic steatosis. Overall, VPA‐triggered over‐regulation of CD36 and DGAT2 could be helpful for a better understanding of the mechanisms underlying VPA‐induced hepatic steatosis and may offer novel therapeutic strategies to combat VPA‐induced hepatotoxicity. HIGHLIGHTSVPA induced hepatic steatosis and modulated genes associated with lipid metabolism.CD36‐mediated fatty acid uptake contributed to VPA‐induced lipid accumulation.VPA increased the hepatic level of DGAT2 through inhibiting MEK‐ERK pathway and enhanced triglyceride synthesis.SREBP‐1c‐mediated fatty acid synthesis was not involved in VPA‐induced hepatic steatosis.

Triterpenoids with Antiplatelet Aggregation Activity from the Roots of Ilex pubescens

Two new triterpenes and five new triterpene saponins, named ilexpusons A-G (1-7), as well as eight known compounds were isolated from Ilex pubescens. The structures of the new compounds were established by a combination of chemical and spectroscopic methods, including HRESIMS, 1H-NMR, 13C-NMR, 1H-1H COSY, HSQC, HMBC, and NOESY. Additionally, the biological activity of compounds 1 - 15 against adenosine diphosphate-induced platelet aggregation in rabbit plasma was determined. Among the tested compounds, 1, 2, 5, 6, 8, 13, 14, and 15 exhibited significant inhibition of platelet aggregation in vitro.

Identification of rotundic acid metabolites after oral administration to rats and comparison with the biotransformation by Syncephalastrum racemosum AS 3.264

The objective of this study was to identify the metabolites of rotundic acid after oral administration to rats and compare the similarities with its biotransformation by Syncephalastrum racemosum AS 3.264 using ultra-high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry. A total of fourteen metabolites were determined based on the mass spectrometry and chromatographic behaviors, among which eleven (M1-M3, M7-M14) and six (M2, M4-M8) metabolites were identified in rats and S. racemosum, respectively. Three identical metabolites (M2, M7 and M8) were found in rats and S. racemosum, indicating that there were metabolic similarities. Moreover, to confirm the results of mass spectrometry, three (M2, M4 and M7) metabolites were obtained by the means of amplifying incubation and their structures were determined by various spectroscopic analyses, and M4 was proved to be a previously undescribed compound. This results showed that in vitro assisted preparation by microbial transformation is a feasible and effective method of obtaining metabolites which are in low amounts and difficult to be prepared in vivo.

A facile and selective approach to the qualitative and quantitative analysis of triterpenoids and phenylpropanoids by UPLC/Q-TOF-MS/MS for the quality control of Ilex rotunda

HIGHLIGHTSA LC–MS method was developed firstly to determine triterpenoids and phenylpropanoids in the five parts of Ilex rotunda.Modified mass defect filter and key product ion filter strategies increased the structural identification efficiency.70 triterpenoids and 12 phenylpropanoids were identified and 48 of them were confirmed by reference substances.12 triterpenoids and 3 phenylpropanoids were simultaneously quantified.PCA and OPLS‐DA analysis were employed to discriminate the origin of stem bark. ABSTRACT Ilex rotunda, in which triterpenoids and phenylpropanoids are major bioactive constituents, has been widely used in traditional Chinese medicines. In this study, a validated UPLC/Q‐TOF‐MS/MS method was developed to simultaneously identify and quantify the triterpenoids and phenylpropanoids in the stem bark, fruit, leaves, roots and stem xylem of this herbal medicine. A total of seventy triterpenoids and twelve phenylpropanoids were identified with the assistance of the modified mass defect filter and key product ion filter data processing strategies, and forty‐eight of them were confirmed by reference substances. Meanwhile, the contents of twelve triterpenoids and three phenylpropanoids in the five plant parts were determined with good linearity (R2≥0.9993), precision (RSD≤2.04%), repeatability (RSD≤1.99%), stability (RSD≤1.88%) and recovery (96.65–103.17% and RSD≤3.54%). Furthermore, PCA and OPLS‐DA methods were employed to visualize the relationships and discrimination of the forty‐two stem bark samples from two origins based on the contents of fifteen analytes. Our findings may provide early scientific evidence for quality control and for elucidating the therapeutic principle of Ilex rotunda.

Inhibition of Glucose-6-Phosphate Dehydrogenase Reverses Cisplatin Resistance in Lung Cancer Cells via the Redox System

The pentose phosphate pathway (PPP), which branches from glycolysis, is correlated with cancer cell proliferation, survival and senescence. In this study, differences in the metabolic profile of the PPP and the redox status of human lung carcinoma A549 cells and cisplatin-induced multidrug-resistant A549/DDP cells were analyzed and evaluated. The results showed that A549/DDP cells exhibited differential PPP-derived metabolic features and redox-related molecules. A549/DDP cells exhibited increased expression and enzymatic activity of PPP enzyme glucose-6-phosphate dehydrogenase (G6PD). Furthermore, as demonstrated by the apoptotic rate, cell viability, and colony formation, inhibition of G6PD by siRNA or an inhibitor sensitized A549/DDP cells to cisplatin. Additionally, inhibition of G6PD restored the cisplatin sensitivity of A549/DDP cells by influencing redox homeostasis. In conclusion, overcoming cisplatin resistance through inhibition of G6PD could improve the understanding of the mechanisms underlying cisplatin-induced resistance in human lung cancer and may provide insights into the therapeutic potential of this treatment to combat resistance.

The therapeutic effect of Ilex pubescens extract on blood stasis model rats according to serum metabolomics

ETHNOPHARMACOLOGICAL RELEVANCE Ilex pubescens Hook. et Arn (MDQ), a traditional Chinese herb, is used in the treatment of cardiovascular diseases. However, the mechanisms underlying the preventive effect of MDQ on blood stasis remain unclear. AIM OF THE STUDY In this study, serum metabolomics integrated with a biochemical assay strategy were established to evaluate the preventive effect and mechanism of action of MDQ on rats with acute blood stasis. MATERIALS AND METHODS Forty-nine rats were divided into seven groups: the control group, model group, aspirin treatment group (30 mg/kg), clopidogrel treatment group (8 mg/kg) and three MDQ treatment groups (250, 500 and 1000 mg/kg). A hybrid quadrupole time of flight mass spectrometry (QTOF/MS) coupled to ultra-high-performance liquid chromatography (UPLC) was applied for profiling the serum metabolites. The multivariate data analysis techniques using unsupervised principal component analysis (PCA) and supervised orthogonal projections to latent structures discriminant analysis (OPLS-DA) were used for pattern recognition and distinguishing variabilities among groups. RESULTS MDQ protected the rats against blood stasis, as evidenced by the restoration of the anti-platelet aggregation activity, fibrinogen concentration, prothrombin time, thrombin time, activated partial thromboplastin time, endothelial nitric oxide synthase, endothelin, thromboxane B2 and 6-keto-prostaglandin F1α. The combination of PCA and OPLS-DA revealed deviations in eighteen differential biomarkers in serum. The identified biomarkers were primarily engaged in the metabolic pathways including arachidonic acid metabolism, glycerophospholipid metabolism, phospholipid biosynthesis and bile acid biosynthesis. The levels of eleven biomarkers showed significant alterations and a tendency to be restored to normal values in MDQ-treated blood stasis rats. Moreover, a correlation network diagram was constructed to show the serum biomarkers perturbed by MDQ. CONCLUSIONS These results suggested that MDQ had preventive effects on blood stasis in rats via arachidonic acid metabolism and glycerophospholipid metabolism.