Comparative Investigation Between Raw and Stir-Frying Processed Cuscutae Semen Based on HPLC Fingerprints Coupled With Chemometric Methods.

BACKGROUND: The curative effect of Cuscutae Semen (CS) could be adjusted after stir-frying, and it is difficult to distinguish between the raw CS and stir-fried processed Cuscutae Semen (SFCS) in clinical practice. OBJECTIVE: The paper aims to establish the fingerprints of raw and fried Cuscutae Semen products by high-performance liquid chromatography (HPLC) so that the compounds in raw and stir-fried products could be analyzed qualitatively and quantitatively. At the same time, the chemometrics method was used to evaluate the difference between CS and SFCS to provide reference for the research of traditional Chinese medicine (TCM) CS. METHOD: The raw and SFCS products' related substances were separated on a C18 column (250 mm × 4.6 mm, 5 µm) maintained at 30 °C (column temperature). The mobile phase comprised 0.1% formic acid aqueous solution (A) and acetonitrile (B), and a detection wavelength of 328 nm. The data were imported into statistical software for chemometric analysis. RESULTS: The developed HPLC method exhibits good linearity and has good systematic applicability. The content of these 12 compounds in the samples was further determined and the data analyzed by chemometrics. The results showed that the composition of CS changed on heating, and HCA showed that CS and SFCS could be clearly distinguished. PCA showed that six components caused differences, namely neochlorogenic acid, cryptophyllogenic acid, caffeic acid, quercetin, isorhamnetin, and kaempferol. CONCLUSIONS: This study unequivocally establishes a chromatographic fingerprint method intended for the extensive analysis of raw and stir-fried processed CS, which could substantially enhance the quality control of CS and the rational development and utilization of TCM resources. HIGHLIGHTS: This method for the simultaneous quantification of multiple compounds in CS and SFCS revealed the components responsible for the differences between raw and processed products. This will provide support for quality control of this herbal medicine.

Study on active ingredients of Jingfang Baidu San for preventing COVID-19 based on network pharmacology and molecular docking / 药学实践杂志

Objective To investigate the active ingredients of Jingfang Baidu San for the prevention and treatment of COVID-19 by using network pharmacology and molecular docking, and to provide references for clinical applications. Methods The chemical constituents and action targets of all medicinal materials in Jingfang Baidu San were retrieved from TCMSP. Uniprot database was used to search the corresponding genes of targets. Cytoscape software was used to construct the network of medicinal materials-compounds-targets for visualization. The target proteins of COVID-19 were searched by disease databases. The intersection of both was considered to be analyzed to establish the protein-protein interaction (PPI) network by STRING database. GO function enrichment analysis and KEGG pathway enrichment analysis were performed through Metascape database to predict its mechanism. The effective strength of core constituents on preventing COVID-19 was calculated by molecular docking method. Results A total of 159 effective ingredients and 277 potential targets were obtained in Jingfang Baidu San within the given screening conditions [oral bioavailability (OB) ≥30%; drug-like (DL) ≥ 0.18], including 55 core targets with the intersection of 273 targets of COVID-19. According to the results of GO and KEGG enrichment analysis performed on the core targets, 1376 GO items and 136 KEGG pathways were obtained, involving infectious diseases, cancer, cell progress, immune system, signaling pathways etc. The results of molecular docking indicated strong binding capacity between the core ingredients and SARS-CoV-2 3CL hydrolase or angiotensin-converting enzyme II (ACE2). The hydrogen binding and hydrophobic effect were the main forms of the interaction. Conclusion The active ingredients in Jingfang Baidu San can inhibit the binding between SARS-CoV-2 protein and ACE2, thus regulating multiple targets and signal pathways, which plays a role in the prevention and the treatment of COVID-19.

Isoliquiritigenin inhibits mouse S180 tumors with a new mechanism that regulates autophagy by GSK-3ß/TNF-α pathway.

In this study, the pharmacokinetic properties and stability of isoliquiritigenin (ILQ) in microsomes were evaluated. The data showed ILQ administrated by i.h had high absorption degree (absolute bioavailability> 90%), and strong elimination ability (average t1/2≈ 67 min). ILQ in rat tissues could reach peak at 0.25 h, and be detected in almost all tissues. In vitro, stability of ILQ in four species liver microsomes were rat > beagle dog > monkey > human > mouse. On the basis of pharmacokinetic (PK) profiles, mechanism of ILQ against S180 was explored. ILQ could not inhibit S180 growth directly in vitro. However, ILQ extremely prohibited S180 tumor volume in vivo. And when TNF-α in NK cells was knocked down by siRNA, ILQ had no inhibiting effect on S180 tumor. ILQ enhanced TNF-α expression in NK cells by FCM detection. Autophagy-associated proteins LC3-II, Beclin-1, ATG-7 were elevated in S180 cells co-cultured with ILQ treating NK cells. When TNF-α was knocked down by siRNA, ILQ could not induce autophagy in S180 tumors. In the NK cells of osteosarcoma patients, TNF-α was negatively correlated with GSK-3ß by ELISA detection. ILQ could inhibit GSK-3ß expression and further increased p65 and c-Rel expression in NK cells. When GSK-3ß was knocked down by siRNA, ILQ did not affect p65 and c-Rel expression. ILQ directly inhibited GSK-3ß and then activated the NF-κB pathway to enhance TNF-α expression in NK cells, which could induce autophagy in sarcomas. The present study supplied a new mechanism for ILQ against tumors.

Effects and primary mechanism of arctigenin in C6 rat glioma / 中国药理学通报

Aim To observe the effect and primary mechanism of arctigenin ( ARG) in C6 rat glioma. At the same time, to investigate the effect of ARG com-bined with temozolomide. Methods C6 glioma rat model was established, and 90 rats were divided into six groups, which were subcutaneously administered with model, low and high ARG (0. 05 and 0. 1 mg· kg-1 , sc) , temozolomide (20 mg·kg-1 , p. o. ) , low ARG combined with temozolomide(TMZ / ARG 0. 05) and high ARG combined with temozolomide ( TMZ /ARG 0. 1 ) . The tumor specimens of brain were col-lected after tumor graft. Proliferating cell nuclear anti-gen ( PCNA ) , glial fibrillary acidic protein ( GFAP ) and CD40 in tumor specimens were determined by im-munohistochemistry. Results ① Compared with the model group, the tumor sizes of rats in the arctigenin treatment groups were decreased ( P significantly decreased PCNA and CD40 expression ( P<0. 05 ) and increased GFAP expression ( P<0. 05 ) .③ Compared with model group, arctigenin combined with temozolomide decreased the tumor sizes ( P <0. 01 ) , and the tumor inhibition rate was higher than that of the arctigenin and temozolomide. At the same time, arctigenin combined with temozolomide de-creased PCNA and CD40 expression ( P <0. 01 ) and increased GFAP expression ( P <0. 05 ) , which was better than arctigenin and temozolomide. Conclusion Arctigenin inhibits rat glioma growth, and synergizes with temozolomide, which may be associated with in-hibiting PCNA and CD40 expression and strengthening GFAP expression.

Inhibitory effect of arctigenin on lymphocyte activation stimulated with PMA/ionomycin / 药学学报

This study investigated the effect of arctigenin (Arc) on the cell activation, cytokines expression, proliferation, and cell-cycle distribution of mouse T lymphocytes. Mouse lymphocytes were prepared from lymph node and treated with Phorbol-12-myristate-13-acetate (PMA)/Ionimycin (Ion) and/or Arc. CD69, CD25, cytokines, proliferation and cell cycle were assayed by flow cytometry. The results showed that, at concentrations of less than 1.00 micromol x L(-1), Arc expressed non-obvious cell damage to cultured lymphocytes, however, it could significantly down-regulate the expression of CD69 and CD25, as well as TNF-alpha, IFN-gamma, IL-2, IL-4, IL-6 and IL-10 on PMA/Ion stimulated lymphocytes. At the same time, Arc could also inhibit the proliferation of PMA/Ion-activated lymphocytes and exhibited lymphocyte G 0/G1 phase cycle arrest. These results suggest that Arc possesses significant anti-inflammatory effects that may be mediated through the regulation of cell activation, cytokines expression and cell proliferation.

Effects of melatonin on cellular viability and injury induced by oxygen and glucose deprivation at different phases of co-cultured neuron and glia / 中国药理学通报

AIM To observe the effects of melatonin on acute and chronic injuries induced by oxygen and glucose deprivation (OGD) in co-cultured neuron and glia and to explore the probable mechanisms of melatonin in antagonizing the injuries. METHODS The injury model of cultured neuron and co-cultured neuron and glia was made by administration of sodium dithionite and glucose-deprived Earles solution. In neuron and glia co-culture, two different models, acute injury model at the phase of OGD and chronic injury model after 'reperfusion' were established. The levels of nitric oxide (NO) and the activity of lactate dehydrogenase (LDH) were measured by Griess reagent and LDH kits respectively. The content of malondialdehyde (MDA) was determined by TBA method. Cell viability was analyzed using colorimetric MTT assay. RESULTS Melatonin increased the level of NO at the concentration of 10 -6 , 10 -7 mol?L -1 and decreased the level of MDA content elevated by OGD at the concentration of 10 -6 , 10 -7 , 10 -8 mol?L -1 in vitro cultured cortical neurons. In the chronic injury model after 'reperfusion' melatonin (10 -6 , 10 -7 , 10 -8 mol?L -1 ) significantly decreased LDH activity and increased MTT value in neurons and glia co-cultured. But in the acute injury model, melatonin obviously increased LDH activity and decreased MTT value. CONCLUSION Melatonin protection for neuron from injuries induced by oxygen and glucose deprivation may be related to increase in the level of NO and decrease in the content of MDA. Melatonin can antagonize the injury in the chronic injury model after 'reperfusion', but exaggerate the injury in the acute injury model. These may be all related to its antioxidant action. Our results also suggest that melatonin may probably inhibit activation of microglia.