Rheum officinale Baill. Treats zebrafish embryo thrombosis by regulating NOS3 expression in the arginine biosynthesis pathway.

BACKGROUND: Rheum officinale Baill. (ROB), as one of the traditional Chinese medicines for promoting blood circulation and removing blood stasis, has a wide range of pharmacological effects, such as cardiovascular protection, and has become a common drug in the clinical care of thrombosis. OBJECTIVE: Although there are some pharmacological studies on ROB in the treatment of thrombotic diseases, the mechanism and material basis are still unclear. Based on the arginine biosynthesis signalling pathway, this research explored the target proteins and metabolites related to the intervention of ROB in thrombosis and expounded on the antithrombotic mechanism of ROB from the comprehensive perspectives of target prediction, intermediate metabolites and potential metabolic pathways. METHODS: In this research, ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) technology was used to qualitatively detect the chemical compounds of ROB, and the antithrombotic activity of ROB was evaluated by establishing a zebrafish model. The target function was predicted by network pharmacology, and differential metabolites were screened by metabolomics and multivariate statistical analysis methods. Correlation analysis of network pharmacology and metabolomics screening results was conducted to identify the potential pathway of ROB intervention in thrombosis, and the prediction results were further verified. RESULTS: ROB significantly reduced the reactive oxygen species (ROS) staining intensity in zebrafish induced by phenylhydrazine (PHZ) and improved the inhibition rate of thrombosis. By constructing the "herb-disease-component-target" network, it was concluded that the active ingredients of ROB in treating thrombosis involved emodin, aloe-emodin and physcion, and the key targets included nitric oxide synthase 2 (NOS2) and nitric oxide synthase 3 (NOS3). A total of 341 differential metabolites in zebrafish with thrombosis were screened by partial least squares discriminant analysis (PLS-DA). The results of reverse transcription-polymerase chain reaction (RT-PCR) experiments and targeted metabolomics verification showed that ROB was mainly involved in improving thrombosis by upregulating the expression of NOS3 mRNA and regulating the levels of arginine, glutamate and glutamine in the arginine biosynthesis pathway. CONCLUSIONS: ROB improved thrombosis by regulating the expression of NOS3 mRNA and the contents of arginine, glutamate and glutamine in the arginine biosynthesis signalling pathway.

Organic anion transporter 1 and 3 contribute to traditional Chinese medicine-induced nephrotoxicity.

With the internationally growing popularity of traditional Chinese medicine (TCM), TCM-induced nephropathy has attracted public attention. Minimizing this toxicity is an important issue for future research. Typical nephrotoxic TCM drugs such as Aristolochic acid, Tripterygium wilfordii Hook. f, Rheum officinale Baill, and cinnabar mainly damage renal proximal tubules or cause interstitial nephritis. Transporters in renal proximal tubule are believed to be critical in the disposition of xenobiotics. In this review, we provide information on the alteration of renal transporters by nephrotoxic TCMs, which may be helpful for understanding the nephrotoxic mechanism of TCMs and reducing adverse effects. Studies have proven that when administering nephrotoxic TCMs, the expression or function of renal transporters is altered, especially organic anion transporter 1 and 3. The alteration of these transporters may enhance the accumulation of toxic drugs or the dysfunction of endogenous toxins and subsequently sensitize the kidney to injury. Transporters-related drug combination and clinical biomarkers supervision to avoid the risk of future toxicity are proposed.

Rheum officinale at different years old stage / 中草药

Objective To analyze the content and variation rules of 10 constituents in radix, rhizome, and leaf of Rheum officinale at one-, two-, and three-year-old stage, respectively, and provide theoretical guidance for efficient production and quality control of the crud drug. Methods The content of each constituent in R. officinale was determined by high performance liquid chromatography (HPLC), and one factor analysis of variance and multiple comparison were performed by SPSS 24.0. Results HPLC system was established for the determination of 10 components in R. officinale. The linear range was good (r2 > 0.997), RSD of precision, stability, and repeatability were less than 2%, and the recoveries were 96.10%—107.10%, respectively. The content analyses showed that, in the same part, the content of gallic acid decreased significantly year by year or at the 2nd growth years (P 3 > 2 (P 1 > 2 (P radix > leaf (P < 0.05). Conclusion The HPLC based determination of 10 constituents in R. officinale showed that the accumulation profiles of the samples at different years or from different parts varied. For the same parts, the contents of most constituents increased year by year. During the same growth year, the contents of most constituents in radix or rhizome were higher than those in leaf. The radix and rhizome of the three years old samples had the highest contents of main constituents.

Affinity chromatographic methodologies based on immobilized voltage dependent anion channel isoform 1 and application in protein-ligand interaction analysis and bioactive compounds screening from traditional medicine.

Voltage dependent anion channel isoform 1 (VDAC-1) serves as an attractive target of anti-cancer drugs by mediating the entry and exit of metabolites between cytoplasm and mitochondria. This work reports on the preparation of a VDAC-1-based bioaffinity chromatographic stationary phase by linking the protein on lecithin modified microspheres. An assay of chromatographic methods including frontal analysis, zonal elution, injection dependent analysis and nonlinear chromatography were utilized to investigate the bindings of ATP, NADH and NADPH to VDAC-1. Electrostatic interactions were found to be main forces during these bindings. The calculated association constants of the three ligands to VDAC-1 showed good agreements between diverse chromatographic methods. Validated application of the stationary phase was performed by screening anti-cancer compounds of Rheum officinale Baill. using high performance affinity chromatography coupled with electrospray ionization-quadrupole time of flight mass spectrometry. Chrysophanol, emodin, rhein, aloe-emodin and catechin were identified as the bioactive components of the herb. These compounds targeted VDAC-1 through Thr207 and the N-terminal region of the protein. Taken together, the current stationary phase was possible to become a promising tool for protein-ligand interaction analysis and anti-cancer drug screening from complex matrices.

Effect of Rheum officinale on activity of intestinal stem cells in Drosophila melanogaster induced by SDS toxicity / 中草药

Objective: To explore the effect of Rheum officinale extracts (ROE) on activity of intestinal stem cells of Drosophila melanogaster. Methods: The control group was fed with normal cornmeal-yeast medium, and the experimental group was fed with cornmeal-yeast medium containing 0.05 or 0.1 g/mL ROE. In the experiment, the gut damage was induced by feeding D. melanogaster with toxic compounds. The effects of ROE on survival rate, number and morphology of progenitor cells, proliferation and differentiation of intestinal stem cells, expression of reactive oxygen species (ROS), number of intestinal epithelial dead cells and life span of D. melanogaster were detected and analyzed. Results: ROE (0.05 and 0.1 g/mL) could increase the survival rate of D. melanogaster induced by toxic compounds. ROE (0.1 g/mL) could decrease SDS-induced ROS levels, reduce the number of intestinal epithelial dead cells, inhibit excessive proliferation and differentiation of intestinal stem cell, alleviate the excessive accumulation of progenitor cells, thereby maintain homeostasis in the gut. In addition, ROE could prolong the lifespan of D. melanogaster. Conclusion: ROE can inhibit excessive proliferation and differentiation of intestinal stem cells, enhance gut immune function, and prolong the life span of D. melanogaster.

Quality control for extracts from aerial parts of Rheum officinale based on multi-assay depending on one determination / 中草药

Objective: To establish a method for the quality control of the extracts from the aerial parts of Rheum officinale (EARO) based on multi-assay depending on one determination (MDOD). Methods: The quality control of EARO was carried out through the thin-layer chromatography(TLC), discrimination of chromatographic peak in fingerprint, ultraviolet spectrophotometric, and HPLC methods. The relative correction factors of aloe-emodin, chrysophanol, and physcione for detecting emodin were set up, the factors were used to measure their contents, and the method was adopted to the quality control of EARO. Results: The experimental and control samples in TLC showed the same colored spots at the same position in EARO, five characteristic peaks among six were identified, and showed a good linear relationship at the range of 4.944-29.664 μg/mL (r = 0.9990). The method of MDOD showed the recoveries of aloe-emodin, emodin, chrysophanol, and physcione were 97.08%, 95.78%, 100.60%, and 97.47%, and the RSD values were 3.32%, 2.42%, 3.72%, and 2.67%. There were no significant differences between MDOD method and external standard method. Conclusion: TLC and fingerprint could be used to identify the chemical constituents in EARO, and ultraviolet spectrophotometry could be used to control the quantity of the total anthraquinone. The method of MDOD could be used to quantitatively control the contents of aloe-emodin, emodin, chrysophanol, and physcione, and the method is simple and accurate, which has a higher repeatability.

Extraction optimization of anthraquinone from aerial part of Rheum officinale Baill based on Plackett-Burman design combined with CCD response surface methodology / 中国药学杂志

OBJECTIVE: To optimize the process of extacting effective constituents from aerial part of Rheum officinale Baill by Plackett-Burman design combined with CCD response surface methodology. METHODS: In Plackett-Burman combined with CCD response surface design, the independent variables were concentration of ethanol, extraction time, and solvent fold. The dependent variables were contents of total anthraquinone and rheum emodin. RESULTS: The optimal extraction process of total anthraquinone was as follows: 67.25% ethanol, 90 min for reflux per time, 28:1 fold of solvent and 2 times of extraction; the optimal extraction process of emodin was 85% ethanol, 90 min for reflux per time, 40 fold of solvent and 2 times of extration. CONCLUSION: The optimal process is simpleand convenient for extracting aerial part of Rheum officinale Baill with high precision and predictability. Copyright 2012 by the Chinese Pharmaceutical Association.