Effects of Levistilide A on Hemorheology and Endothelial Cell Injury in Rats with Blood Stasis

Background. Vascular endothelial cell injury is not only the initiating factor of cardiovascular and cerebrovascular diseases but also the essence of blood stasis. Levistilide A (LA), a natural component isolated from the traditional Chinese herb, Ligusticum chuanxiong Hort, has traditional effects on improving blood circulation and removing stasis. In this study, the effects and potential mechanisms of LA in the rat model of blood stasis and the mechanism in endothelial cell injury have been explored. Materials and Methods. In this experiment, the effects of LA on the model of acute blood stasis in rats were explored. The blood samples were collected for the measurement of coagulation and hemorheological indices, and the carotid arteries were also excised from rats for hematoxylin-eosin (HE) staining and immunohistochemistry (IHC). In addition, the improvement effects of LA on the H2O2-induced human umbilical vein endothelial cell (HUVEC) injury model were evaluated. And the cell viability detection was conducted by the CCK8 assay, and the pathway-related protein expressions were detected by western blotting. Results. In vivo, compared with the model group, the treatment of LA (10 mg/kg) could reduce the FIB (fibrinogen) content ( P < 0.01 ), increase the INR (international normalized ratio) and PT (prothrombin time) ( P < 0.01 ), and reduce the plasma viscosity ( P < 0.05 ) and whole blood viscosities of low, medium, and high shear rates in the blood of blood stasis model rats ( P < 0.01 ). In vitro, the cell viability in the LA-pretreated group was higher than that of the model group ( P < 0.05 ). The expression levels of PI3K, AKT, and eNOs in the LA-pretreated group were increased ( P < 0.01 ) as compared to the model group. Conclusion. These findings demonstrated that LA has the ability to improve blood hypercoagulation and blood viscosity, and enhance the viability of cells. It is more likely that it exerts a protective effect on the endothelial cell through the PI3K-AKT-eNOs pathway. These results indicate LA will be a potential candidate to cure blood stasis with endothelial cell injury.

Interactions of thymic stromal lymphopoietin with interleukin-4 in adaptive immunity during Aspergillus fumigatus keratitis

AIM: To investigate the potential interactions of thymic stromal lymphopoietin (TSLP) with interleukin-4 (IL-4) in adaptive immunity during fungal keratitis (FK). METHODS: An FK mouse model was induced with Aspergillus fumigatus (AF) hyphal infection. Mice were divided into several groups: untreated, phosphate buffer saline (PBS), infected with AF, and pretreated with a scrambled siRNA, a TSLP-specific siRNA (TSLP siRNA), murine recombinant TSLP (rTSLP), immunoglobulin G (IgG), murine recombinant IFN (rIFN-γ), murine recombinant IL-4 (rIL-4), rIL-13, murine recombinant IL-17A (rIL-17A), and murine recombinant IL-17F (rIL-17F) groups. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) or Western blot were performed to determine mRNA and protein levels in the inflamed cornea. Cytokine locations were observed by immunofluoresence staining after AF hyphal infection. RESULTS: Compared to those in the untreated group, TSLP and T helper type 1 (Th1) cytokine levels in the AF group were upregulated at 24h post infection (hpi), and those of T helper type 2 (Th2) and T helper type 17 (Th17) cytokines were increased at 5d post infection (dpi). Th2 cytokine levels were decreased in the TSLP siRNA-pretreated group and increased in the rTSLP-pretreated group compared with the AF group. The TSLP level was increased in the rIL-4-pretreated group, but there were no significant changes among the other groups. Immunofluorescence staining showed cytokine locations after AF hyphal infection. CONCLUSION: TSLP induces a Th2 immune response and promots Th2 T cell differentiation in vivo. IL-4 promotes TSLP secretion. Therefore, TSLP with IL-4 regulates adaptive immunity in FK.

Effect of the beta-adrenergic blockade on intestinal lactate production and glycogen concentration in dogs infused with hexoses

Objectives To investigate effect of beta adrenergic blockade on intestinal lactate production and glycogen concentration in dogs infused with hexoses. Methods Experiments were carried out on 35 fasted male anaesthetized dogs weighing between 9 and 16 kg. The animals were divided into 7 (5 dogs per group) groups. Group I dogs served as control and infused with normal saline, groups II-IV were intravenously infused with glucose (1.1 mg/kg/min), fructose (1.1 mg/kg/min) and galactose (1.1 mg/kg/min) respectively while groups V-VII animals were pretreated with propranolol (0.5 mg/kg) and were infused with glucose, fructose or galactose respectively. A vein draining the proximal segment of the jejunum was cannulated along with right and left femoral arteries and veins. Glucose uptake was calculated as the product of jejunal blood flow and the difference between arterial and venous glucose levels (A-V glucose), part of the jejunum tissue was homogenized for estimation of glycogen concentration, and plasma lactate was assayed using lactate colorimetric kit. Results The result showed significant increase in venous lactate production in response to glucose (78.30 ± 4.57 mg/dl), fructose (60.72 ± 1.82 mg/dl) and galactose (71.70 ± 1.30 mg/dl) when compared with the control group (51.75 ± 1.32 mg/dl) at (p<0.05) with no significant difference in animals pretreated with propranolol. There was no significant difference in glycogen concentration (p>0.05) in animals infused with hexoses only compared with propanolol pretreated group. Conclusions Results suggests that one of the possible fates of the enormous amount of glucose taken up by the intestine is conversion to lactate and not glycogen and β-adrenergic receptor does not affect it.

Simultaneous Pretreatment of Aspirin and Omega-3 Fatty Acid Attenuates Nuclear Factor-κB Activation in a Murine Model with Ventilator-Induced Lung Injury

Ventilator-induced lung injury (VILI) is an important critical care complication. Nuclear factor-κB (NF-κB) activation, a critical signaling event in the inflammatory response, has been implicated in the tracking of the lung injury. The present study aimed to determine the effect of simultaneous pretreatment with enteral aspirin and omega-3 fatty acid on lung injury in a murine VILI model. We compared the lung inflammation after the sequential administration of lipopolysaccharides and mechanical ventilation between the pretreated simultaneous enteral aspirin and omega-3 fatty acid group and the non-pretreatment group, by quantifying NF-κB activation using an in vivo imaging system to detect bioluminescence signals. The pretreated group with enteral aspirin and omega-3 fatty acid exhibited a smaller elevation of bioluminescence signals than the non-pretreated group (p = 0.039). Compared to the non-pretreated group, the pretreatment group with simultaneous enteral aspirin and omega-3 fatty acid showed reduced expression of the pro-inflammatory cytokine, tumor necrosis factor-α, in bronchoalveolar lavage fluid (p = 0.038). Histopathological lung injury scores were also lower in the pretreatment groups compared to the only injury group. Simultaneous pretreatment with enteral administration of aspirin and omega-3 fatty acid could be a prevention method for VILI in patients with impending mechanical ventilation therapy.

Effect of Hydroalcoholic Extract of Satchys Lavandulifolia on Pentylenetetrazole-Induced Seizures in Male Mice: The Role of Gabaergic and Opioidergic Systems

Introduction: Epilepsy is one of the most common neurological disorders. Though there are effective medications available for treatment of epilepsy, the use of most drugs is associated with many side effects and drug interactions. Stachys lavandulifolia (SL) used in Iranian traditional medicine show anti-anxiety and sedative actions. The objective of the current study was to evaluate the anticonvulsant effect of hydroalcoholic extract of SL on the pentylenetetrazole (PTZ)-induced seizure in male mice and the role of benzodiazepine and opioid receptors. Methods: This study was conducted on 100 male mice randomly categorized into 10 groups: Normal Saline, Diazepam groups (0.025 and 0.1 mg/kg), SL extract groups (50, 100 and 200 mg/kg), Diazepam 0.025 mg/kg + SL extract 50mg/kg and three groups that pre-treated with NS, Flumazenil or Naloxone, 5 min before injection of 200 mg/kg extract. After 30 min, PTZ (80 mg/kg) was injected to animals and seizure indices were evaluated. Results: The SL extract attenuated the PTZ-induced seizures in a dose dependent manner and pre-treatment with flumazenil reversed this effect of SL extract but pre-treatment with naloxone could not reverse this effect, because seizure indices on naloxone pretreated group was still lower than normal saline group. Combination of ineffective dose of diazepam and SL extract decrease PTZ-induced seizures. Discussion: The results of our study showed the anticonvulsant properties of hydroalcoholic extract of SL. These effects might be due to the impact of the components of this extract on the central benzodiazepine system.

Agronomical, physiological and molecular evaluation reveals superior salt-tolerance in bread wheat through salt-induced priming approach

Salt stress significantly limit wheat crop productivity worldwide. Exposure to non-lethal levels of salt stress, referred to as "salt-priming", allows plants to persist subsequent lethal conditions; the priming effect continues even after an extended salt stress-free period. This study attempted to evaluate the effectiveness of the salt-induced priming approach to cope with the toxic effects of long-term salinity stress in wheat. After 22 days of gradual salt acclamation to reach 250 mM NaCl, plants were recovered for eight days and finally shocked with 250 mM NaCl (priming+shock) for 7 days. After that, physiological parameters and gene expression of six salt-responsive genes were assessed. Additionally, 120 days after germination (at the end of the season), agronomic traits were recorded. Analysis of the agronomical traits revealed higher productivity in the salt-pretreated group (priming+shock) plants than the non-pretreated (shock only). Consistently, salt-pretreated plants maintained higher photosynthetic pigments level and decreased proline and MDA content than non-pretreated, suggesting enhanced salt tolerance. Moreover, salt-pretreated plants sustained high expressional levels of salt-responsive genes (TaNHX1, TaSOS1, TaSOS4, TaHKT1, TaHKT2, and TaAKT1) comparing with non-pretreated, indicating a vital role in ion homeostasis and conferring salt tolerance. Ultimately, this finding could facilitate novel smart approaches to improve wheat productivity under salt stress.

Bevacizumab and Sorafenib Modulate P-Glycoprotein Function In Vitro and Bevacizumab Increases In Vivo Sorafenib Plasma Concentrations in Mice

Overexpression of P-glycoprotein (P-gp) is associated with multidrug resistance. Since sorafenib (NEXAVAR®) is a P-gp (an efflux protein of ATP-binding cassette family) substrate, we tested whether bevacizumab (AVASTIN®), a monoclonal antibody directed toward VEGF (Vascular Endothelial Growth Factor) and sorafenib could modulate P-gp functionality. In vitro two human ovarian carcinoma cells (IGROV1) overexpressing or weakly expressing P-gp were used. Bevacizumab and sorafenib effects on P-gp functionality were evaluated by measuring doxorubicin intracellular accumulation. In vivo study was to document whether bevacizumab could modify sorafenib disposition in mice. Therefore, concentrations of sorafenib were determined by HPLC in plasma of mice bearing a human colorectal carcinoma xenograft when sorafenib is given orally (5 mg/kg) on day 4, alone or after a pretreatment with bevacizumab (5 mg/kg IP) on days 1 and 3. In vitro a significant doxorubicin accumulation and reversion of doxorubicin resistance in P-gp expressing cell lines were observed with bevacizumab or sorafenib pretreatment In vivo, sorafenib AUC was 1.44 fold significantly higher in bevacizumab pretreated group and Cmax was 1.35 fold higher in bevacizumab-pretreated group. Mean residence time of sorafenib increased in the presence of bevacizumab, this increase reflects an improvement of sorafenib bioavailability after bevacizumab pretreatment. We may conclude that bevacizumab pretreatment decreases P-gp functionality and increases doxorubicin intracellular accumulation in vitro and sorafenib plasma concentrations in vivo.

A unique radioprotective effect of resolvin E1 reduces irradiation-induced damage to the inner ear by inhibiting the inflammatory response

Background In addition to the direct effects of irradiation, the induced inflammatory response may play an important role in the damage to the inner ear caused by radiotherapy for the treatment of head and neck cancers. Resolvin E1 has anti-inflammatory activity, acting by reducing neutrophil infiltration and proinflammatory cytokine expression. Therefore, in this study we sought to confirm whether the inflammation induced by irradiation was involved in damage to the inner ear after radiotherapy and to investigate the protective effect and underlying mechanism of resolvin E1 using mouse models. Methods A dose of resolvin E1 was delivered by intraperitoneal injection to mice before irradiation. Changes in the auditory brainstem response, relative balance ability, inner ear morphology and the expression levels of proinflammatory cytokines in the inner ear were analyzed on days 7 and 14 after irradiation and compared among three experimental groups. Results Morphological analysis of the inner ear showed severe damage to the cochlea and vestibule after irradiation. In contrast, damage to the cochlea and vestibule was significantly reduced in the resolvin E1-pretreated group, which nonetheless remained damaged compared to the wild-type group. Proinflammatory cytokines also showed expression patterns matching the morphological observations. Conclusions We believe that the inflammation induced by irradiation is involved in the damage to the inner ear caused by radiotherapy, and that resolvin E1 reduces the damage caused by irradiation to the inner ear by regulating the induced inflammatory response.

The effects of carnosine pretreatment on the inflammatory response and the PI3K/Akt signaling pathway following hypoxia-ischemia in neonatal rats

An increasing number of studies have demonstrated that carnosine plays a neuroprotective role in many types of brain injury. We have previously shown that carnosine has both short-term and long-lasting neuroprotective effects in a hypoxia–ischemia(HI) rat model. In the mature brain, post-ischemia neuronal survival involves in activation of the phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway, whether the activation of PI3K/Akt pathway also plays an important role in the immature brain still remain unclear.The goal of this study is to detect the effection of carnosine on inflammation response following HI, further evidencing neuroprotection of carnosine. We measured total Akt, phospho-Akt (p-AKT) and tumor necrosis factor receptor 1 (TNFR1) protein levels by western blot assay and tumor necrosis factor-α (TNF-α) and TNFR1 mRNA expression using real-time RT-PCR. We found the carnosine-pretreated group had statistically significant downregulation of TNF-α mRNA levels 24 h after HI (P < 0.05). Similar results were observed when we measured TNFR1 mRNA levels both 24h and 72h after HI (P < 0.05). And the TNFR1 protein expression after HI was markedly decreased at 24 and 72 h post-HI in the carnosine-pretreated rats(P < 0.05). Nevertheless, the rats pretreated with carnosine showed a marked increase in p-Akt levels (P< 0.05). And the pro-apoptotic protein Bad was also examined using immunohistochemistry after 24 and 72 h of all groups. We found significantly fewer Bad-positive cells in the carnosine-pretreated group at each time point after HI (P < 0.05). These findings suggest that carnosine pretreatment inhibits the HI-induced inflammatory response, and neuroprotection mechanism of carnosine involved in activation of the PI3K/Akt signaling pathway.

Effect of Ticagrelor, a Cytochrome P450 3A4 Inhibitor, on the Pharmacokinetics of Tadalafil in Rats

Tadalafil is a cytochrome P450 (CYP) 3A4 substrate. Because there are few data on drug-drug interactions, it is advisable to take sufficient consideration when co-administering tadalafil with CYP3A4 inducers or inhibitors. This study was conducted to assess the effect of ticagrelor, a CYP3A4 inhibitor, on the pharmacokinetic properties of tadalafil after oral administration to rats. A total of 20 Sprague–Dawley male rats were randomly divided into the non-pretreated group and ticagrelor-pretreated group, and tadalafil was orally administered to each group after pretreatment with or without ticagrelor. Blood samples were collected at predetermined time points after oral administration of tadalafil. As a result, systemic exposure of tadalafil in the ticagrelor-pretreated group was significantly increased compared to the non-pretreated group (1.61-fold), and the clearance of tadalafil in the ticagrelor-pretreated group was significantly reduced than the non-pretreated group (37%). The prediction of the drug profile through the one-compartment model could explain the differences of pharmacokinetic properties of tadalafil in the non-pretreated and ticagrelor-pretreated groups. This study suggests that ticagrelor reduces a CYP3A-mediated tadalafil metabolism and that tadalafil and a combination regimen with tadalafil and ticagrelor requires dose control and specific pharmacotherapy.