The Anxiolytic Effect of Polysaccharides from Stellariae Radix through Monoamine Neurotransmitters, HPA Axis, and ECS/ERK/CREB/BDNF Signaling Pathway in Stress-induced Male Rats.

BACKGROUND: Stellaria dichotoma L. var. lanceolata Bge. is renowned for its efficacy in "clearing deficiency heat" and represents a significant traditional Chinese medicine (TCM) resource. Modern pharmacology has demonstrated the anti-anxiety effects of Stellaria dichotoma L. var. lanceolata Bge. polysaccharides (SDPs). SDPs are one of the active constituents of Stellaria dichotoma L. var. lanceolata Bge. This study presents the first extraction of SDPs and investigates their potential molecular mechanisms and anxiolytic effects that are not previously reported. METHODS: First, SDPs were obtained by water extraction and alcohol precipitation and analyzed for their monosaccharide composition by high performance liquid chromatography (HPLC). Male SD rats were subjected to a two-week indeterminate empty bottle stress procedure and a three-day acute restraint stress procedure, during which diazepam (DZP) (1 mg/kg) and SDPs (50, 100 and 200 mg/kg, intragastrically) were administered. A number of behavioral tests, including the elevated plus maze test (EPM), the open field test (OFT) and the light/dark box test (LDB), were used to assess the anti-anxiety potential of SDPs. Serum levels of Corticosterone (CORT) and Adrenocorticotropic hormone (ACTH), as well as the levels of Dopamine (DA) and serotonin (5-HT) found in the hippocampus and frontal cortex, were quantified using commercially available enzyme-linked immunosorbent assay (ELISA) kits. In addition, protein levels of key proteins cAMP-response element binding protein (CREB), phospho-CREB (p-CREB), brain-derived neurotrophic factor (BDNF), ERK½, p-ERK½, and GAPDH expression in rat hippocampus were measured by Western blot analysis, and modulation of the endocannabinoid system was assessed by immunohistochemistry. RESULTS: Following administration of SDPs (50, 100, 200 mg/kg) and diazepam 1 mg/kg, anxiolytic activity was exhibited through an increase in the percentage of arm opening times and arm opening time of rats in the elevated plus maze. Additionally, there was an increase in the number of times and time spent in the open field center, percentage of time spent in the open box, and shuttle times in the LDB. Furthermore, tissue levels of DA and 5-HT were increased in the hippocampus and frontal cortex of rats after treatment with SDPs. In addition, SDPs significantly decreased serum levels of CORT and ACTH in rats. SDPs also effectively regulated the phosphorylation of the extracellular regulated protein kinases (ERK) and CREB-BDNF pathway in the hippocampus. Moreover, the expression levels of CB1 and CB2 proteins were heightened due to SDPs treatment in rats. CONCLUSIONS: The study verified that SDPs alleviate anxiety in the EBS and ARS. The neuroregulatory behavior is accomplished by regulating the Monoamine neurotransmitter, HPA axis, and ECB-ERK-CREB-BDNF signaling pathway.

Simultaneous quantification of pirarubicin, doxorubicin, cyclophosphamide, and vincristine in human plasma of patients with non-Hodgkin's lymphoma by LC-MS/MS method.

Pirarubicin (THP), doxorubicin (DOX), cyclophosphamide (CTX), and vincristine (VCR) are widely used in the treatment of patients with non-Hodgkin's Lymphoma. Herein, a precise and sensitive method was developed for the determination of THP, DOX, CTX and VCR in human plasma by high-performance liquid-chromatography-tandem mass spectrometry (LC-MS/MS). Liquid-liquid extraction was applied to extract THP, DOX, CTX, VCR, and the internal standard (IS, Pioglitazone) in plasma. Agilent Eclipse XDB-C18 (3.0 mm × 100 mm) was utilized and chromatographic separation was obtained in eight minutes. Mobile phases were composed of methanol and buffer (10 mM ammonium formate containing 0.1% formic acid). The method was linear within the concentration range of 1-500 ng/mL for THP, 2-1000 ng/mL for DOX, 2.5-1250 ng/mL for CTX, and 3-1500 ng/mL for VCR. The intra- and inter-day precisions of QC samples were found to be below 9.31 and 13.66%, and accuracy ranged from -0.2 to 9.07%, respectively. THP, DOX, CTX, VCR and the internal standard were stable in several conditions. Finally, this method was successfully utilized to simultaneously determine THP, DOX, CTX and VCR in human plasma of 15 patients with non-Hodgkin's Lymphoma after intravenous administration. Finally, the method was successfully employed in the clinical determination of THP, DOX, CTX, and VCR in patients with non-Hodgkin lymphoma after administration of RCHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine, and prednisone) regimens.

Dichotomine B Attenuates Neuroinflammatory Responses by Regulating TLR4/MyD88-mTOR Signaling Pathway in BV2 Cells.

Dichotomine B is a [Formula: see text]-Carboline alkaloid isolated from Stellariae Radix. Stellariae Radix, also known as Yin Chai Hu, is a common Chinese medicine in clinical practice. This herb has been demonstrated to have anti-inflammatory activity. This study aimed to investigate the effects and mechanisms of Dichotomine B on neuroinflammation by BV2 microglia induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP). The experiment was divided into a control group, a model group (10 µg/mL LPS + 5 mM ATP), a model+ TLR4 inhibitor (TAK-242, 10 µmol/L) group, model+ Dichotomine B (20, 40 and 80 µmol/L) groups and Dichotomine B (80 µmol/L) group. The BV2 cell viability was detected by MTT assay, the morphology of BV2 cells was observed by inverted microscope, and the levels of IL-6, IL-1[Formula: see text] and TNF-[Formula: see text] in BV2 cells were determined by ELISA. The expression levels of TLR4, MyD88, p-mTOR/mTOR, p62, p-RPS6/RPS6, LC3II/LC3I and Beclin-1 proteins were detected by western blot assay. The expression levels of TLR4, MyD88, mTOR, p62, RPS6, LC3B and Beclin-1 mRNA were detected by PCR assay. Finally, molecular docking was performed to predict the affinity of Dichotomine B with TLR4, MyD88 and mTOR by LibDock of Discovery Studio and MOE. The results showed that compared with the model group, the survival rates of damaged cells were significantly increased by TAK-242 and Dichotomine B, and the morphology of these BV2 cells improved. The levels of IL-6, IL-1[Formula: see text] and TNF-[Formula: see text] were significantly decreased by TAK-242 and Dichotomine B in LPS/ATP-induced BV2 cells. 80 µmol/L Dichotomine B has no effect on normal BV2 cells. Further mechanism investigation showed that TAK-242 and Dichotomine B significantly inhibited the protein and mRNA expression levels of TLR4, MyD88, p-mTOR/mTOR (mTOR), p62, p-RPS6/RPS6 (RPS6) and increased the protein and mRNA expression levels of LC3II/LC3I (LC3B), Beclin-1. Docking study showed the LibDock scores of Dichotomine B with TLR4, MyD88 and mTOR were all higher than those of positive drugs (Diazepam). These findings indicated that Dichotomine B attenuated neuroinflammatory responses in LPS/ATP-induced BV2 microglia, and its mechanism may be related to TLR4/MyD88-mTOR signaling pathway and autophagy.

Betaine Inhibits NLRP3 Inflammasome Hyperactivation and Regulates Microglial M1/M2 Phenotypic Differentiation, Thereby Attenuating Lipopolysaccharide-Induced Depression-Like Behavior.

Depression is one of the most important mental illnesses and is closely related to inflammation. Betaine is a natural product with an anti-inflammatory and antioxidant activities. However, the mechanism by which betaine ameliorates depression-like behaviors induced by lipopolysaccharide (LPS) is poorly understood. The purpose of this study was to investigate the neuroprotective effect of betaine on LPS-induced depression-like behavior in mice and its mechanism of action. ICR mice were randomly divided into four groups: the control group, the LPS model group (0.83 mg/kg), the positive drug group (MIDO, 50 mg/kg), and the betaine group (5% and 1% in drinking water). The betaine group was administered for 21 days, and on the 22nd day, except for the blank group, LPS (0.83 mg/kg) was intraperitoneally injected to establish a lipopolysaccharide-induced mice depression-like model. Twenty-four hours after LPS injection, the tail suspension test (TST), open field test (OFT), and sucrose preference test (SPT) were performed to evaluate the effect of betaine on LPS-induced depressive behavior in mice. After the behavioral study, the mouse brain, hippocampus, and serum were taken for detection. The expressions of cytokines and inflammatory mediators were detected by ELISA, HE staining, immunofluorescence, immunohistochemistry, and western blotting. Western blotting was used to detect the protein expression levels of the nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, and ASC, the protein expression levels of the microglial polarization markers COX-2, inducible nitric oxide synthase (iNOS), and CD206. The results showed that betaine significantly ameliorated the depression-like behavior in LPS-induced mice, significantly attenuated the production of proinflammatory cytokines and increased the release of an anti-inflammatory cytokines. Betaine decreased the expression of the NLRP3 inflammasome, decreased the expression of M1 polarization markers, tumor necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), COX-2, and iNOS and promoted the expression of M2 polarization marker CD206. Our study suggests that betaine may promote the transition of microglia from the M1 to the M2 phenotype by inhibiting NLRP3 inflammasome activation, thereby attenuating lipopolysaccharide-induced depression-like behavior.

The traditional uses, phytochemistry, pharmacology and toxicology of Dictamnus dasycarpus: a review.

OBJECTIVES: Dictamnus dasycarpus is a plant of the Rutaceae family, and its root bark is the main part used as a medicine, named 'Bai-Xian-Pi'. It is used to clear away heat, remove dampness, and dispel wind and also used for detoxification. The purpose of this review is to provide a systematic review about the botany, traditional uses, phytochemistry, pharmacology and toxicology of this plant. KEY FINDINGS: More than 200 compounds have been isolated and identified from the plant, including alkaloids and their glycosides, terpenoids and their derivatives and phenylpropanoids. Extensive pharmacological activities of the extracts or compounds of D. dasycarpus in vivo and in vitro were mainly confirmed, including anti-inflammatory activity, protecting cardiovascular activity, improving liver injury and anti-cancer activity. SUMMARY: In this paper, the botany, traditional uses, phytochemistry and pharmacology of D. dasycarpus were reviewed. In the future, D. dasycarpus needs further study, such as paying more attention to quality control and the utilization on agriculture. In addition, discussing the medicinal components of decoction as well as the toxicity will also contribute to the progress of clinical trial studies.

A review of the ethnobotanical value, phytochemistry, pharmacology, toxicity and quality control of Tussilago farfara L. (coltsfoot).

ETHNOPHARMACOLOGICAL RELEVANCE: Tussilago farfara L. (commonly called coltsfoot), known as a vital folk medicine, have long been used to treat various respiratory disorders and consumed as a vegetable in many parts of the world since ancient times. AIM OF THE REVIEW: This review aims to provide a critical evaluation of the current knowledge on the ethnobotanical value, phytochemistry, pharmacology, toxicity and quality control of coltsfoot, thus provide a basis for further investigations. MATERIALS AND METHODS: A detailed literature search was obtained using various online search engines (e.g. Google Scholar, Web of Science, Science Direct, Baidu Scholar, PubMed and CNKI). Additional information was sourced from ethnobotanical literature focusing on Chinese and European flora. The plant synonyms were validated by the database 'The Plant List' (www.theplantlist.org). RESULTS: Coltsfoot has diverse uses in local and traditional medicine, but similarities have been noticed, specifically for relieving inflammatory conditions, respiratory and infectious diseases in humans. Regarding its pharmacological activities, many traditional uses of coltsfoot are supported by modern in vitro or in vivo pharmacological studies such as anti-inflammatory activities, neuro-protective activity, anti-diabetic, anti-oxidant activity. Quantitative analysis (e.g. GC-MS, UHPLC-MRMHR) indicated the presence of a rich (>150) pool of chemicals, including sesquiterpenes, phenolic acids, flavonoids, chromones, pyrrolizidine alkaloids (PAs) and others from its leaves and buds. In addition, adverse events have resulted from a collection of the wrong plant which contains PAs that became the subject of public concern attributed to their highly toxic. CONCLUSIONS: So far, remarkable progress has been witnessed in phytochemistry and pharmacology of coltsfoot. Thus, some traditional uses have been well supported and clarified by modern pharmacological studies. Discovery of therapeutic natural products and novel structures in plants for future clinical and experimental studies are still a growing interest. Furthermore, well-designed studies in vitro particularly in vivo are required to establish links between the traditional uses and bioactivities, as well as ensure safety before clinical use. In addition, the good botanical identification of coltsfoot and content of morphologically close species is a precondition for quality supervision and control. Moreover, strict quality control measures are required in the studies investigating any aspect of the pharmacology and chemistry of coltsfoot.

Effects of Betaine on LPS-Stimulated Activation of Microglial M1/M2 Phenotypes by Suppressing TLR4/NF-κB Pathways in N9 Cells.

Microglia mediate multiple facets of neuroinflammation. They can be phenotypically divided into a classical phenotype (pro-inflammatory, M1) or an alternative phenotype (anti-inflammatory, M2) with different physiological characteristics and biological functions in the inflammatory process. Betaine has been shown to exert anti-inflammatory effects. In this study, we aimed to verify the anti-inflammatory effects of betaine and elucidate its possible molecular mechanisms of action in vitro. Lipopolysaccharide (LPS)-activated microglial cells were used as an inflammatory model to study the anti-inflammatory efficacy of betaine and explore its mechanism of regulating microglial polarisation by investigating the morphological changes and associated inflammatory changes. Cytokine and inflammatory mediator expression was also measured by ELISA, flow cytometry, immunofluorescence, and western blot analysis. Toll-like receptor (TLR)-myeloid differentiation factor 88 (Myd88)-nuclear factor-kappa B (NF-κB) p65, p-NF-κB p65, IκB, p-IκB, IκB kinase (IKK), and p-IKK expression was determined by western blot analysis. Betaine significantly mitigated the production of pro-inflammatory cytokines and increased the release of anti-inflammatory cytokines. It promoted the conversion of the microglia from M1 to M2 phenotype by decreasing the expression of inducible nitric oxide synthase and CD16/32 and by increasing that of CD206 and arginase-1. Betaine treatment inhibited the TLR4/NF-κB pathways by attenuating the expression of TLR4-Myd88 and blocking the phosphorylation of IκB and IKK. In conclusion, betaine could significantly alleviate LPS-induced inflammation by regulating the polarisation of microglial phenotype; thus, it might be an effective therapeutic agent for neurological disorders.

Quality evaluation for Radix Astragali based on fingerprint, indicative components selection and QAMS.

Radix Astragali (RA) is one of the most widely used Chinese herbs prescribed in many Chinese formulas to reinforce 'Qi' and treat vital energy deficiency. This study combined fingerprinting with quantitative analysis multi-components by a single marker (QAMS) to improve the quality control standard for RA on the basis of existing quality control methods of traditional Chinese medicinal materials. UPLC-ESI-TOF-MS technique was used to evaluate the quality of RA by fingerprinting and QAMS. Using the anti-inflammatory, anti-oxidation and anti-anoxic activities to screen characteristic components of RA, the calycosin-7-O-ß-d-glucoside (CG), ononin, astragaloside IV, astragaloside II, calycosin and astrageloside I significantly inhibited ear edema in mice, the calycosin and CG had good antioxidant activity and the astragaloside I had a significant anti-hypoxia activity. Astragaloside I, astragaloside II, astragaloside IV, ononin, calycosin and CG had significant pharmacological actions. These components were comprehensively used as the indicative components for the quality control of RA. Astragaloside I was used as the internal standard of the relative correction factors of CG (13.45), ononin (0.51), calycosin (12.08), astragaloside IV (0.73) and astragaloside II (0.81). Astragaloside I and CG were used as internal standards of the relative correction factors of the flavonoids and saponins of ononin (1.11), calycosin (0.04), astragaloside IV (0.73) and astragaloside II (0.81). The study combined fingerprinting with QAMS to improve the quality control standard for RA.

Anti-inflammatory effect of Calycosin glycoside on lipopolysaccharide-induced inflammatory responses in RAW 264.7 cells.

Objective to find the Calycosin glycoside (CG) potential anti-inflammatory effect. The anti-inflammatory activity of CG was evaluated by lipopolysaccharide (LPS)-induced RAW 264.7 cells, and the IL-6, IL-1ß, TNF-α and PGE2 level were measured by ELISA; NO levels in the culture media were determined using the Griess reaction. The iNOS and COX-2 mRNA expressions were measured by qRT-PCR and the phosphorylation of IκBα, p65, ERK, JNK, and p38 was determined by western blotting. The CG could markedly inhibit the productions of NO and PGE2 and markedly inhibit the productions of TNF-α, IL-1ß and IL-6. CG could suppress mRNA expression of iNOS, COX-2 and protein phosphorylation of IκBα, p65, ERK, JNK, and p38 in LPS-induced RAW 264.7 cells. These findings indicate that CG exhibited potent anti-inflammatory activity through the NF-κB and MAPK signal pathway, and as a potential therapeutic agent against inflammatory diseases.

Ethnopharmacology, phytochemistry, and pharmacology of Cornus officinalis Sieb. et Zucc.

ETHNOPHARMACOLOGICAL RELEVANCE: Cornus officinalis (Cornaceae), known in Chinese as "Shanzhuyu," is a frequently used traditional Chinese medicine. It tastes sour and is astringent and slightly warm in nature. Its fruits have long been used to treat kidney deficiency, high blood pressure, waist and knee pain, dizziness, tinnitus, impotence, spermatorrhea, menorrhagia, and other diseases in China. The main distribution areas are Shanxi and Gansu. AIM OF THE STUDY: This review focused on the ethnopharmacological uses of the herb. We also focus on the phytochemical, pharmacological, and toxicological studies on C. officinalis. The recent analytical methods developed for the quality control of the herb's constituents are also reviewed. Additionally, future trends and prospects in the study of this herb are proposed. MATERIALS AND METHODS: Information on C. officinalis was gathered by searching the internet (PubMed, ScienceDirect, Wiley, ACS, CNKI, Scifinder, Web of Science, Google Scholar, and Baidu Scholar) and libraries. RESULTS: This review compiled the ethnopharmacological uses, including the classic prescriptions and historical applications. Approximately 300 chemical compounds have been isolated and identified from C. officinalis. The major active components of the plant are organic acids and iridoids, among which morroniside and loganin have been extensively investigated. The fruit of the plant has been used in treating many diseases in traditional medicine. Scientific studies indicated the herb's wide range of pharmacological activities, such as hepatic and renal protection, antidiabetes activity, cardioprotection, antioxidation, neuroprotection, antitumor activity, anti-inflammation, analgesic effects, antiaging activity, antiamnesia, antiosteoporosis, and immunoregulation. The analytical methods developed for the quantitative and qualitative determination of various compounds in the herb were further reviewed. CONCLUSIONS: In this paper, we reviewed various studies conducted on C. officinalis, especially in areas of its ethnopharmacological use, as well as on its phytochemistry, pharmacology, and modern analytical methods used. Some of the herb's ethnomedical indications have been confirmed by the herb's pharmacological effects, such as its hepatic and renal protection and the antidiabetic effects. In particular, the crude extract and its chemical composition have exerted good therapeutic effect in diabetic treatment. C. officinalis entails additional attention on its pharmacological effects and drug development to expand its effective use clinically. Many advanced technologies are used for quality testing, but the detection component is exceedingly scarce for synthetically evaluating the quality of C. officinalis herbs. Thus, further research is necessary to investigate the quality control and toxicology of the plant, to further elucidate its clinical use, and to control herbal quality.

Hepatoprotective Effects of Kaempferol-3-O-α-l-Arabinopyranosyl-7-O-α-l-Rhamnopyranoside on d-Galactosamine and Lipopolysaccharide Caused Hepatic Failure in Mice.

Fulminant hepatic failure (FHF), associated with high mortality, is characterized by extensive death of hepatocytes and hepatic dysfunction. There is no effective treatment for FHF. Several studies have indicated that flavonoids can protect the liver from different factor-induced injury. Previously, we found that the extracts of Elaeagnus mollis leaves had favorable protective effects on acute liver injury. However, the role and mechanisms behind that was elusive. This study examined the hepatoprotective mechanisms of kaempferol-3-O-α-l-arabinopyranosyl-7-O-α-l-rhamnopyra-noside (KAR), a major flavonol glycoside of E. mollis, against d-galactosamine (GalN) and lipopolysaccharide (LPS)-induced hepatic failure. KAR reduces the mouse mortality, protects the normal liver structure, inhibits the serum aspartate aminotransferase (AST) and alamine aminotransferase (ALT) activity and decreases the production of malondialdehyde (MDA) and reactive oxygen species (ROS) and inflammatory cytokines, TNF-α, IL-6, and IL-1ß. Furthermore, KAR inhibits the apoptosis of hepatocytes and reduces the expression of TLR4 and NF-κB signaling pathway-related proteins induced by GalN/LPS treatment. These findings suggest that the anti-oxidative, anti-inflammatory, and anti-apoptotic effects of KAR on GalN/LPS-induced acute liver injury were performed through down-regulating the activity of the TLR4 and NF-κB signaling pathways.

[Macroscopic and microscopic identification of Ligularia przewalskii].

OBJECTIVE: To provide macroscopic and microscopic identification basis for Ligularia przewalskii. METHODS: Macroscopic and microscopic identification of roots, stems and leaves of Ligularia przewalskii were carried out with the methods of paraffin section, leaves epidermal section and powder transdermal section. RESULTS: The microscopic characteristics included: Open collateral vascular bundles in stem were not in the same size and arranged in two rings; Lots of fiber bundles scattered in the column parts; There were two vascular bundles in principal vein of leaf; Anticlinal wall of upper epidermis cells was thickened like moniliform, lower epidermis were like waves with irregular; The type of stoma was anomocytic; Calcium oxalate acicular crystal could be seen in the powder. CONCLUSION: These features can provide references for identification of Ligularia przewalskii.