EETs Attenuate Ox-LDL-Induced LTB4 Production and Activity by Inhibiting p38 MAPK Phosphorylation and 5-LO/BLT1 Receptor Expression in Rat Pulmonary Arterial Endothelial Cells.

Cytochrome P-450 epoxygenase (EPOX)-derived epoxyeicosatrienoic acids (EETs), 5-lipoxygenase (5-LO), and leukotriene B4 (LTB4), the product of 5-LO, all play a pivotal role in the vascular inflammatory process. We have previously shown that EETs can alleviate oxidized low-density lipoprotein (ox-LDL)-induced endothelial inflammation in primary rat pulmonary artery endothelial cells (RPAECs). Here, we investigated whether ox-LDL can promote LTB4 production through the 5-LO pathway. We further explored how exogenous EETs influence ox-LDL-induced LTB4 production and activity. We found that treatment with ox-LDL increased the production of LTB4 and further led to the expression and release of both monocyte chemoattractant protein-1 (MCP-1/CCL2) and intercellular adhesion molecule-1 (ICAM-1). All of the above ox-LDL-induced changes were attenuated by the presence of 11,12-EET and 14,15-EET, as these molecules inhibited the 5-LO pathway. Furthermore, the LTB4 receptor 1 (BLT1 receptor) antagonist U75302 attenuated ox-LDL-induced ICAM-1 and MCP-1/CCL2 expression and production, whereas LY255283, a LTB4 receptor 2 (BLT2 receptor) antagonist, produced no such effects. Moreover, in RPAECs, we demonstrated that the increased expression of 5-LO and BLT1 following ox-LDL treatment resulted from the activation of nuclear factor-κB (NF-κB) via the p38 mitogen-activated protein kinase (MAPK) pathway. Our results indicated that EETs suppress ox-LDL-induced LTB4 production and subsequent inflammatory responses by downregulating the 5-LO/BLT1 receptor pathway, in which p38 MAPK phosphorylation activates NF-κB. These results suggest that the metabolism of arachidonic acid via the 5-LO and EPOX pathways may present a mutual constraint on the physiological regulation of vascular endothelial cells.

Effects of the inhalation of the m3 receptor antagonist bencycloquidium bromide in a mouse cigarette smoke-induced airway inflammation model.

Bencycloquidium bromide (BCQB), a novel M3 receptor antagonist, alleviates airway hyperresponsiveness, inflammation, and airway remodeling in a murine model of asthma. The aim of this study was to investigate the anti-inflammatory activity of inhaled BCQB in a cigarette smoke (CS)-induced model of acute lung inflammation. Mice exposed to CS developed chronic obstructive pulmonary disease (COPD). Inhalation of BCQB suppressed the accumulation of neutrophils and macrophages in airways and lung and also inhibited the CS-induced increases in mRNA levels of keratinocyte-derived chemokine, monocyte chemotactic protein-1, tumor necrosis factor-alpha, and interleukin-1ß in lung and protein expression levels in bronchoalveolar lavage fluid. Moreover, BCQB (300 µg/ml) inhibited the CS-induced changes in superoxide dismutase and myeloperoxidase activities in the lungs. Our study suggests that BCQB might be a potential therapy for inflammation in CS-induced pulmonary diseases, including COPD.

Pharmacokinetics of aconitine-type alkaloids after oral administration of Fuzi (Aconiti Lateralis Radix Praeparata) in rats with chronic heart failure by microdialysis and ultra-high performance liquid chromatography-tandem mass spectrometry.

ETHNOPHARMACOLOGICAL RELEVANCE: Fuzi [the lateral root of Aconitum carmichaeli Debx (Ranunculaceae)] is a well-known traditional medicinal herb used to treat chronic heart failure (CHF). Aconitine-type alkaloids are major alkaloids that are responsible for the pharmacological activity and toxicity of this herb.To investigate therapeutic effects and pharmacokinetic profiles of aconitine-type alkaloids in CHF rats. MATERIALS AND METHODS: The plasma pharmacokinetic profiles of aconitine, mesaconitine, and hypaconitine were investigated after once treatment of Fuzi extract (containing aconitine 0.086 mg/g, mesaconitine 0.84 mg/g, and hypaconitine 1.97 mg/g) using a rapid and sensitive combinative method of ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and microdialysis (MD). The cardiac function and antioxidant enzyme activities were also evaluated. RESULTS: Recoveries of MD sampling ranged from 35.06% to 45.74% with RSD below 6.05%. Fuzi extract improved the myocardial function and antioxidant enzymatic activities of rats with CHF. Aconitine, mesaconitine, and hypaconitine exhibited slower absorption into the bloodstream, and yielded 11-fold less values of area under concentration-time curve (AUC) in the CHF rats than those in normal rats. The plasma AUC showed that the maximum blood concentration (Cmax) was 5.561 ng/mL for aconitine, 17.30 ng/mL for mesaconitine, and 17.78 ng/mL for hypaconitine in normal rats, while these were 0.6059 ng/mL, 2.430, and 0.7461 ng/mL in CHF rats, respectively. CONCLUSION: Aconitine-type alkaloids associated with Fuzi׳s efficacy have lower intake and slower elimination in the CHF rats, indicating a non-interdependent relationship between its efficacy and toxicity. It may contribute to the depth understanding of the toxicological and pharmacological profiles of Fuzi and further benefit the herbal drug development with safety and efficacy for CHF treatment.

Anti-fatigue and vasoprotective effects of quercetin-3-O-gentiobiose on oxidative stress and vascular endothelial dysfunction induced by endurance swimming in rats.

Chronic fatigue accumulation increases the incidence of cardiovascular disease while the treatment of antioxidants could prevent this development. We have previously shown that quercetin-3-O-gentiobiose (QG), a flavonoid isolated from tonic herb Okra, possesses anti-oxidative properties. In the present study, the protective effects of QG were evaluated in a rat model of load-induced endurance swimming. Oral administration of QG at the doses of 25-75mg/kg could significantly improve the endurance capability of rats to fatigue along with decrease serum lactic acid and blood urea nitrogen levels were decreased. Moreover, QG could alleviate vascular impairments, enhance the activities of antioxidant enzymes and attenuate the levels of inflammatory cytokines (MCP-1, IL-6 and TNF-α). The results indicated that QG had anti-fatigue and vasoprotective effects and represented a potential agent for the treatment of aortic pathology involved with fatigue- and related syndrome.

Antitumor and immunomodulatory effects of a water-soluble polysaccharide from Lilii Bulbus in mice.

Lilii Bulbus is a popular folk medicine in the worldwide and has attracted great attention due to its bioactivity against respiratory system diseases (include lung cancers). This study was the first report providing in vivo evidences of antitumor potential of the bioactive polysaccharide from Lilii Bulbus. One major fraction (LBP-1) was obtained by purifying the crude polysaccharides extracted from Lilii Bulbus. Chemical characterization analysis indicated that LBP-1 was only a glucan, whose average molecular weight was 30.5 kDa. Intraperitoneal administration of LBP-1 at the doses of 50-200mg/kg significantly inhibited the growth of Lewis lung carcinoma. Moreover, it could also obviously increase macrophage phagocytosis, splenocytes proliferation and cytokine (TNF-α, IL-2, IL-6 and IL-12) production to participate in the antitumor effects. LBP-1 could act as antitumor agent with immunomodulatory activity.

[Textural research on "Xuelihua" in "Bencao Gangmu Shiyi" (A Supplement to the Compendium of Materia Medica)].

To investigate the historical origins of folk herbal medicine 'Xuelihua', the origin of 'Xuelihua' was confirmed by textural, botanical characteristics research, investigation of therapeutic effects and field survey. The 'Xuelihua' which was originally recorded in 'Bencao Gangmu Shiyi' (A Supplement to the Compendium of Materia Medica) should be 'Riben Shegencao' (Ophirrhiza japonica) of Rubiaceae family.

Fatty acids in tea shoots (Camellia sinensis (L.) O. Kuntze) and their effects on the growth of retinal RF/6A endothelial cell lines.

Chemo-protective effects of tea on ocular diseases were recorded in Chinese pharmacopoeia about 2000 years ago by eating tea. In the present study, contents of fatty acids (FAs) in tea shoots were determined by capillary GC; and the growth of RF/6A cells was also investigated by exposure to various representative FAs existing in tea shoots with pathologically relevant concentrations (40-500 microM) by ameliorated MTT assay and flow cytometry. Electron spin resonance (ESR) was used to measure oxygen consumption and investigate the free radical scavenging ability of linoleic acid (LA). Results showed that the most abundant long chain FAs were palmitic, linoleic, and alpha-linolenic acid in tea shoots; some RF/6A cells became suspended in culture medium treated by a high dose of both saturated and unsaturated FAs, but no apoptosis was observed. Moreover, it seemed that those FAs with different structure had various effects on the cell proliferation at their relatively low concentrations, LA expressed antioxidant activity in this study, which might be an important mechanism on the protection of eyes.

Cytotoxicity of epigallocatechin-3-gallate to LNCaP cells in the presence of Cu2+.

Epigallocatechin-3-gallate (EGCG) has shown remarkably anti-cancer activity, with its bioactivity being related to reactive conditions, such as pH and metal ions. The present study investigated the degradation of EGCG and its effect on prostate cancer cell in the presence of Cu2+. EGCG was incubated with prostate cancer cells, LNCaP, pretreated with or without Cu2+. EGCG in F-12 medium was quantified using HPLC and the viability of cells was assessed by gel electrophoresis, flow cytometry, and electron microscope. The results of HPLC showed that EGCG degraded completely within 12 h in F-12 medium with or without Cu2+. Gel electrophoresis and flow cytometry did not detect apoptosis of LNCaP cells when they were incubated with EGCG. Electron microscopy examination revealed that EGCG-Cu2+ complex led to damage of cytoplasm membrane in LNCaP cells. It was speculated that not EGCG, but its oxide and complex with Cu2+, are the bioactive components responsible for its cytotoxicity to LNCaP prostate cancer cells.