ABSTRACT
Rapeseed (Brassica napus L.) is susceptible to nutrient stresses during growth and development; however, the CPA (cation proton antiporter) family genes have not been identified in B. napus and their biological functions remain unclear. This study was aimed to identify the molecular characteristics of rapeseed CPAs and their transcriptional responses to multiple nutrient stresses. Through bioinformatics analysis, 117 BnaCPAs, consisting of three subfamilies: Na+/H+ antiporter (NHX), K+ efflux antiporter (KEA), and cation/H+ antiporter (CHX), were identified in the rapeseed genome. Transcriptomic profiling showed that BnaCPAs, particularly BnaNHXs, were transcriptionally responsive to diverse nutrient stresses, including Cd toxicity, K starvation, salt stress, NH4+ toxicity, and low Pi. We found that the salt tolerance of the transgenic rapeseed lines overexpressing BnaA05.NHX2 was significantly higher than that of wild type. Subcellular localization showed that BnaA05.NHX2 was localized on the tonoplast, and TEM combined with X-ray energy spectrum analysis revealed that the vacuolar Na+ concentrations of the BnaA05.NHX2-overexpressing rapeseed plants were significantly higher than those of wild type. The findings of this study will provide insights into the complexity of the BnaCPA family and a valuable resource to explore the in-depth functions of CPAs in B. napus.
Subject(s)
Brassica napus , Brassica rapa , Brassica napus/genetics , Antiporters/genetics , Protons , Brassica rapa/genetics , Vacuoles , Gene Expression Regulation, Plant , Stress, PhysiologicalABSTRACT
The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.
ABSTRACT
The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.
ABSTRACT
Objective To investigate whether ROS/JNK/Egr-1 signaling pathway was activated in cardiomyocytes after hypoxia/reoxygenation ( H/R).Methods H9c2 cardiomyocytes were grouped randomly as follows: control group, H/R group, control +the ROS donor xanthine oxidase /hypoxanthine (XO/HX) group, H/R +the ROS scavenger edaravones (EDA) group, H/R +the ROS scavenger N-acetyl-L-cysteine (NAC) group, H/R +JNK inhibitor SP60012 group.To establish H9c2 H/R models and the myocardial cells were treated with different concentrations of EDA (2 × 10 -6,2 ×10 -5,2 ×10 -4 M), NAC (5 ×10 -4,2 ×10 -3,8 ×10 -3 M), XO/HX (1mU/mL/1.2 ×10 -4 M , 3mU/mL/3.6 ×10 -4 M, 5mU/mL/6.0 × 10 -4 M) and SP600125 (2 ×10 -5 M).ROS level was measured by flow cytometry, and Egr-1, p-JNK and total JNK protein levels were detected by Western blot.ResuIts ROS levels and Egr-1 protein levels in H/R group were significantly higher than control group (P<0.05).The moderate and high concentrations EDA and NAC of ROS scavenger significantly decreased the high levels of ROS and Egr-1 protein ( P<0.05 ) , but there were no significant differences of low concentration.There was a significant positive correlation between ROS levels and Egr-1 protein (r=0.91,P<0.01).JNK activation levels in each concentrations of XO/HX were significantly higher than control group, and JNK activation increased with the increasing of XO/HX concentrations (P<0.05).JNK activation level in H/R group was higher than control group, after treated by EDA and NAC of ROS scavenger and JNK inhibitor, JNA activation reduced (P<0.05).Egr-1 protein levels in H/R group was higher than that in control group, and JNK inhibitor reduced the expression of Egr-1 protein induced by H/R.ConcIusion H/R activates ROS/Egr-1 signaling pathway in H9c2 cardiomyocytes, and JNK activation plays an important role in this pathway.
ABSTRACT
A laboratory experiment was conducted to study effects of urea fertilizer on the chemical composition of soil solutions over time, and to determine Al toxicity as a function of rates of urea application. The experiment revealed that addition of urea fertilizer to soils caused drastic changes in soil pH during the hydrolysis and nitrification stages of urea transformation in the experiment. These pH changes, depending on the N rate of urea application and time courses, had variable effects on soil exchangeable Al, extracted with artificial solutions containing 1 mol l(-1) KCl. The Al mobilization rate could be resolved into two phases: A declining phase for Al was attributed to the urea-induced hydrolysis while a second rising phase was dependent with the nitrification of added N fertilizer. The decreases in exchangeable Al reached the greatest in 4-7 days after fertilization, consistent with soil pH increase. Decreased Al availability had been observed as a consequence of increasing urea addition and soil pH when using Root elongation of maize seedlings as the estimators. Results from the present study demonstrate that urea fertilizer to the surface of soils may lead to a temporary immobilization of Al and, therefore, alleviated Al toxicity to plant seedlings.
