To discover new inhibitors on tissue factor procoagulant activity, 21 tetrahydroprotoberberines were screened on the model of human THP-1 cells stimulated by lipopolysaccharide. Among these tetrahydroprotoberberines, several unique compounds were synthesized through microbial transformation: compound 6 (l-corydalmine) was obtained through regio-selective demethylation by Streptomyces griseus ATCC 13273, whereas compounds 4a, 4b, 5h, and 5i were microbial glycosylation products by Gliocladium deliquescens NRRL1086. The bioassay results showed that compounds 3 (tetrahydroberberine), 10 (tetrahydroberberrubine), and 5f (cinnamyl ester of 5) and 5i (glycosidic product of 5), exhibited the most potential effects, with IC50 values of 8.35, 6.75, 3.75, and 8.79 nM, respectively. The preliminary structure and activity relationship analysis revealed that the 2,3-methylenedioxy group of the A ring was essential for the strong inhibitory effects, and the R configuration of the chiral center C-14 showed higher activity than S-form products. The formation of fatty acid or aromatic acid esters of compound 5, except the cinnamyl esters, would weaken its effects. It is also interesting to note that the glycosylation of tetrahydroprotoberberines will maintain and even enhance the inhibitory effects. Because of the importance of glycochemistry in new drug discovery and development, this deserves further exploration and may provide some guide on the semi-synthesis of tetrahydroprotoberberines as tissue factor pathway inhibitors. Our findings also provide some potential leading compounds for tissue factor-related diseases, such as cancer and cardiovascular diseases.A series of THPBs derivatives were prepared by chemical and microbial semi-synthesis and screened for their inhibitory effect on TF procoagulant activity in human THP-1 cells stimulated by LPS.

BackgroundConsidering the key role of TF in coagulation of sepsis or acute lung injury (ALI), we investigated whether berberine (BBR) could inhibit TF expression and procoagulant activity and explored its possible mechanism.MethodsThe effects of berberine on the expression, procoagulant activity of TF and related signal pathways induced by lipopolysaccharide (LPS) were observed in THP-1 cells.ResultsOur results showed that berberine could inhibit LPS-induced TF activity and expression, and down-regulate NF-κB, Akt and MAPK/JNK/p38/ERK pathways.ConclusionBerberine inhibits TF expression and related pathway, which provides some new insights on its mechanism for sepsis treatment.

The principle of “same treatment for different diseases” is the major reflection of treatment determination based on pathogenesis obtained through the differentiation of symptoms and signs in traditional Chinese medicine (TCM). It is of great importance to explore such principles from the viewpoint of TCM prescriptions, so as to propagate and develop TCM theory. Danggui-Shaoyao-San (DSS), a famous TCM complex prescription first recorded in the book of Jin Kui Yao Lue, has been widely used in the treatment for various diseases in China. Previous reference mining showed that its pathogenesis linked with “liver depression and spleen deficiency,” and there were a lot of similarities between pharmacological activities of DSS and the physiological and pathological function of histamine. This study suggested that DSS significantly inhibited local skin vascular permeability and remarkably decreased the scratching number and time, decreased locomotive activities increased by histamine, and exerted analgesia activities via the H1 receptor. A hypothesis has been proposed that DSS can modulate histamine H1 receptor expression and the relevant signal transduction pathway so as to treat various diseases, while “liver depression and spleen deficiency” would be related to the dysfunction of the histaminergic neuron system, whose regulating center is considered to be the histamine neurons located in the posterior hypothalamus based on a great deal of previous studies, casual findings, and international academic progress. Further investigation is under way to explore the principles or mechanism of DSS for treating different diseases, which may provide some new ideas or methods for the fundamental research of Integrated Chinese and Western Medicine. It provides some reference for reasonable and normative application of DSS in the clinic, accumulates new data for enriching scientific connotation of “liver depression and spleen deficiency” and the pathological mechanism of related diseases, and consequently academically enhances the TCM development.

Acute lung injury is still a significant clinical problem with a high mortality rate and there are few effective therapies in clinic. Here, we studied the inhibitory effect of ruscogenin, an anti-inflammatory and anti-thrombotic natural product, on lipopolysaccharide (LPS)-induced acute lung injury in mice basing on our previous studies. The results showed that a single oral administration of ruscogenin significantly decreased lung wet to dry weight (W/D) ratio at doses of 0.3, 1.0 and 3.0 mg/kg 1 h prior to LPS challenge (30 mg/kg, intravenous injection). Histopathological changes such as pulmonary edema, coagulation and infiltration of inflammatory cells were also attenuated by ruscogenin. In addition, ruscogenin markedly decreased LPS-induced myeloperoxidase (MPO) activity and nitrate/nitrite content, and also downregulated expression of tissue factor (TF), inducible NO synthase (iNOS) and nuclear factor (NF)-κB p-p65 (Ser 536) in the lung tissue at three doses. Furthermore, ruscogenin reduced plasma TF procoagulant activity and nitrate/nitrite content in LPS-induced ALI mice. These findings confirmed that ruscogenin significantly attenuate LPS-induced acute lung injury via inhibiting expressions of TF and iNOS and NF-κB p65 activation, indicating it as a potential therapeutic agent for ALI or sepsis.Download full-size imageHighlights► Ruscogenin markedly suppressed LPS-induced lung W/D ratio and histopathological changes at oral doses of 0.3–3.0 mg/kg. ► Ruscogenin decreased LPS-induced nitrate/nitrite content, myeloperoxidase activity in the lung tissue at three doses. ► Ruscogenin effectively decreased LPS-induced expression of tissue factor, iNOS and p-p65 in the lung tissue at three doses. ► Ruscogenin significantly reduced plasma nitrate/nitrite content and TF procoagulant activity in LPS-induced ALI mice.

Oxidative stress is a major cause in neurodegenerative diseases including Alzheimer's disease (AD), Parkinson's disease (PD), and cerebral ischemia. Ginsenoside Rg1, a natural product extracted from Panax ginseng C.A. Meyer, has been reported to exert notable neuroprotective activities, which partly ascribed to its antioxidative activity. However, its molecular mechanism against oxidative stress induced by exogenous hydrogen peroxide (H2O2) remained unclear. In this study, we investigated its effect on H2O2-induced cell death and explored possible signaling pathway in PC12 cells. We proved that pretreatment with Rg1 at concentrations of 0.1–10 μM remarkably reduced the cytotoxicity induced by 400 μM of H2O2 in PC12 cells by MTT and Hoechst and PI double staining assay. Of note, we demonstrated the activation of NF-κB signaling pathway induced by H2O2 thoroughly in PC12 cells, and Rg1 suppressed phosphorylation and nuclear translocation of NF-κB/p65, phosphorylation and degradation of inhibitor protein of κB (IκB) as well as the phosphorylation of IκB-kinase complex (IKK) by western blotting or indirect immunofluorescence assay. Besides, Rg1 also inhibited the activation of Akt and the extracellular signal-regulated kinase 1/2 (ERK1/2). Furthermore, the protection of Rg1 on H2O2-injured PC12 cells was attenuated by pretreatment with two NF-κB pathway inhibitors (JSH-23 or BOT-64). In conclusion, our results suggest that Rg1 could rescue the cell injury by H2O2 via down-regulation NF-κB signaling pathway as well as Akt and ERK1/2 activation, which put new evidence on the neuroprotective mechanism of Rg1 against the oxidative stress and the regulatory role of H2O2 in NF-κB pathway in PC12 cells.Graphical abstractDownload full-size imageResearch highlights▶ Ginsenoside Rg1 protected against PC12 cells death injured by H2O2. ▶ Ginsenoside Rg1 inhibited the activation of the NF-κB pathway and the nuclear translocation of NF-κB/p65 induced by H2O2. ▶ Ginsenoside Rg1 inhibited the activation of Akt and ERK1/2 induced by H2O2. ▶ The neuroprotective activity of Rg1 against oxidative stress induced by H2O2 was relative with the direct and indirect regulation of the NF-κB activation.

Diosgenin (Dio), a major active component of steroidal sapogenin of the traditional Chinese herb Dioscorea zingiberensis C.H.Wright, shows various activities including anti-inflammatory, anti-thrombotic activities, anti-cancer properties etc. In the present study, we found that diosgenin significantly suppressed the phosphorylation of lung NF-κB p50/p65 and MAPK/p38 in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice, when given orally at doses of 0.1, 1.0 and 10 mg/kg 1 h prior to LPS challenge (30 mg/kg, intravenous injection). Moreover, diosgenin attenuated the lung histopathological changes such as pulmonary edema, coagulation and infiltration of inflammatory cells. In addition, diosgenin significantly decreased the lung wet to dry weight (W/D) ratio and nitrite/nitrate content at three doses, and also markedly inhibited LPS-induced body temperature decrease and nitrite/nitrate elevation in plasma. Besides, diosgenin could significantly suppress activation of NF-κB p65/p50, p38 and expression of inducible nitric oxide synthase (iNOS) in LPS-induced THP-1 cells. Our findings indicate the potential application of diosgenin for ALI treatment.Download full-size imageHighlights► Diosgenin inhibited LPS-induced activations of lung NF-κB p50/p65 and MAPK/p38 at oral doses of 0.1, 1 and 10 mg/kg. ► Diosgenin markedly suppressed LPS-induced lung W/D ratio and histopathological changes at three doses. ► Diosgenin remarkably reduced lung and plasma nitrate/nitrite content in LPS-induced ALI mice. ► Diosgenin significantly suppress LPS-induced decreased body temperature at three doses. ► Diosgenin inhibited LPS-induced activations of NF-κB p50/p65 and MAPK/p38 and iNOS expression at 1 μM in THP-1 cells.

•Pulmonary endothelial barrier dysfunction is involved in a variety of high mortality diseases.•MLCK, RhoA, calcium, tyrosine kinases, apoptosis are major barrier-disruptive factors.•Rac1, Cdc42, S1P, cAMP, ARs, PAR1, FoxM1 are major barrier-protective factors.The confluent pulmonary endothelium plays an important role as a semi-permeable barrier between the vascular space of blood vessels and the underlying tissues, and it contributes to the maintenance of circulatory fluid homeostasis. Pulmonary endothelial barrier dysfunction is a pivotal early step in the development of a variety of high mortality diseases, such as acute lung injury (ALI). Endothelium barrier dysfunction in response to inflammatory or infectious mediators, including lipopolysaccharide (LPS), is accompanied by invertible cell deformation and interendothelial gap formation. However, specific pharmacological therapies aiming at ameliorating pulmonary endothelial barrier function in patients are still lacking. A full understanding of the fundamental mechanisms that are involved in the regulation of pulmonary endothelial permeability is essential for the development of barrier protective therapeutic strategies. Therefore, this review summarizes several important molecular mechanisms involved in LPS-induced changes in pulmonary endothelial barrier function. As for barrier-disruption, the activation of myosin light chain kinase (MLCK), RhoA and tyrosine kinases; increase of calcium influx; and apoptosis of the endothelium lead to an elevation of lung endothelial permeability. Additionally, the activation of Rac1, Cdc42, protease activated receptor 1 (PAR1) and adenosine receptors (ARs), as well as the increase of cyclic AMP and sphingosine-1-phosphate (S1P) content, protect against LPS-induced lung endothelial barrier dysfunction. Furthermore, current regulatory factors and strategies against the development of LPS-induced lung endothelial hyper-permeability are discussed.

Aim of the studyThe analgesic and anti-inflammatory activities of the ethanol extract of Aquilaria sinensis (Lour.) Gilg. Leaves were observed in various experimental models related to nociception and inflammation, so as to provide some evidence for its traditional use.Materials and methodsAcetic acid-induced writhing and a hot plate test in mice were used to evaluate its analgesic activity. On the other hand, its anti-inflammatory activity was observed in xylene or carrageenan-induced edema, carboxymethylcellulose sodium (CMC-Na)-induced leukocyte migration in mice and lipopolysaccharide (LPS)-induced nitric oxide (NO) release from mouse peritoneal macrophages in vitro.ResultsThe ethanol extract significantly inhibited acetic acid-induced writhing after single oral administration at doses of 424 and 848 mg extract/kg, and the response to the thermal stimulus in mice at the dose of 848 mg/kg. Meanwhile, the ethanol extract also remarkably lessened xylene-induced ear swelling, carrageenan-induced paw edema, and CMC-Na-induced leukocyte migration. Furthermore, the extract considerably reduced NO release from LPS-stimulated macrophages with IC50 of 80.4 mg/ml.ConclusionThese findings suggest that Aquilaria sinensis (Lour.) Gilg. Leaves extract present notable analgesic and anti-inflammatory activities, which support its folkloric use for some diseases related with painful and inflammatory conditions such as trauma etc.

Ethnopharmacological relevanceYiQiFuMai Powder Injection (YQFM) is a modern preparation derived from Sheng-mai San, a traditional Chinese prescription, widely used for the treatment of cardiovascular and cerebrovascular diseases. However, its potential molecular mechanism remains unclear.Aim of the studyThe present study was designed to observe the effect of YQFM on oxygen-glucose deprivation (OGD)-induced the brain microvascular endothelial barrier dysfunction and to explore the underlying pathways in vitro.MethodsA mouse brain microvascular endothelial cell line (bEnd.3) was subjected to OGD (2–9 h) to examine the efficacy and molecular mechanisms in the presence or absence of YQFM (100, 200 and 400 μg/ml).ResultsThe results of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and Trans-endothelial electrical resistance (TEER) assays demonstrated that treatment with YQFM increased the cell viability and TEER value, decreased even blue (EB) albumin leakage after OGD in bEnd.3 cells. Western blotting and immunofluorescence staining showed that YQFM reduced the breakage and translocation of Zonula occludens-1 (ZO-1) and claudin-5 after 4 h of OGD and decreased the expression of these proteins after 9 h of OGD. Moreover, YQFM significantly inhibited the expression, phosphorylation and nuclear translocation of NF-κB/p65 and decreased the expression of intercellular adhesionmolecule-1 (ICAM-1) and cyclooxygenase (COX-2) as well as production of nitric oxide (NO). In addition, real time-PCR results revealed that YQFM suppressed the mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) after 4 h of OGD. Furthermore, YQFM markedly inhibited both the phosphorylation of myosin light chain (MLC) and cytoskeletal reorganization and reduced the expression of cleaved-ROCK1 in bEnd.3 cells subjected to OGD.ConclusionThese findings suggest that YQFM ameliorates the OGD-induced brain microvascular endothelial cell barrier disruption associated with the NF-κB/p65 and ROCK1/MLC signaling pathways. These data provide new insights into the use of this preparation for treating cerebrovascular diseases.Download high-res image (269KB)Download full-size image