Tissue kallikrein protects cortical neurons against hypoxia/reoxygenation injury via the ERK1/2 pathway

Systemic or local delivery of human tissue kallikrein gene (hTK) has been shown to be an effective strategy to alleviate cerebral ischemia/reperfusion (I/R) injury, and tissue kallikrein (TK) administration can suppress glutamate- or acidosis-mediated neurotoxicity in vitro. In the present study, the role of TK in hypoxia/reoxygenation (H/R) induced neuronal cell death was investigated. We found that TK administration could remarkably alleviate H/R-induced neuronal injury by reduction of LDH release and promotion of neuron viability. The protective effects of TK could be counteracted by bradykinin B2 receptor (B2R) antagonist HOE140, which could suppress up-regulation of TK on the ERK signal pathway under H/R condition. These results indicate that TK plays an important role in preventing neurons from H/R damage at least partially through the TK-B2R-ERK1/2 pathway.     

Betulinic acid inhibits cell proliferation,migration, and inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes

Betulinic acid (BA), a pentacyclic triterpene derived from the bark of the white birch tree, has been reported to have a variety of pharmacological effects, including antioxidant, anti-inflammatory, antitumor, immunomodulatory, and antiarthritis properties. However, the role of BA in rheumatoid arthritis (RA) remains unclear. Thus, the objective of this study was to examine the effects of BA on RA fibroblast-like synoviocytes (RA-FLS) proliferation, migration, and inflammatory response, and further explore the potential underlying mechanisms. Our results showed that BA inhibited the proliferation, migration, and invasion of RA-FLSs. BA also attenuated tumor necrosis factor-α (TNF-α), enhanced matrix metalloproteinases (MMPs) expression, and inflammatory cytokines production in RA-FLS. Furthermore, BA prevented the activation of Akt/NF-κB pathway in RA-FLS exposed to TNF-α. In conclusion, these findings indicated that BA inhibits cell proliferation, migration, and inflammatory response in RA-FLS; and the Akt/NF-κB signaling pathway was involved in the protective effect of BA on RA-FLS. Thus, BA might be a potential therapeutic agent for the treatment of RA.     

缺氧复氧对滑膜细胞IGF-IGFBP-3 以及线粒体的影响

目的:探讨氧波动环境对原代成纤维样滑膜细胞(fibroblast-like synoviocyte, FLS)胰岛素样生长因子-1(insulin growth factor-1,IGF-1)、胰岛素样生长因子结合蛋白-3(insulin-like growth factor binding protein-3, IGFBP-3)及线粒体的影响。方法:分离并鉴定正常人滑膜细胞,再对滑膜细胞进行分组:对照组、缺氧/再充氧(hypoxia/reoxygenation,H/R)干预组。采用实时定量PCR检测滑膜细胞中IGF-1、IGFBP-3的m RNA水平;Western blot检测滑膜细胞中IGF-1、IGFBP-3的蛋白水平;流式细胞仪检测线粒体膜电位(Mitochondrial membrane potential, MMP)以及线粒体通透性转换孔(Mitochondrial Permeability Transition Pore, MPTP)的变化。结果:与对照组比较,H/R干预组的相对IGF-1和IGFBP-3的m RNA水平和蛋白表达水平显著升高(P0.05),膜电位水平降低(P0.05),线粒体通透性转换孔开放。结论:氧波动环境可促进IGF-1和IGFBP-3的表达及细胞线粒体损伤,其可能是骨关节炎(OA)发病的重要机制之一。     

Involvement of K+ channel-dependant pathways in lipoxin A4-induced protective effects on hypoxia/reoxygenation injury of cardiomyocytes

Studies have shown that lipoxin A₄ (LXA₄) and activation of LXA₄ receptor provided protection against myocardial ischemia/reperfusion injury in animal models. However, the mechanisms by which LXA₄ induced protective role on myocardial ischemia/reperfusion injury remains unclear. In the present studies, we investigated the protective effects of LXA₄ on H9c2 cardiomyocytes exposed to hypoxia/reoxygenation (H/R) injury and involvement of heme oxygenase-1 (HO-1)- and K⁺ channel-dependant pathways in the LXA₄ action. H9c2 cardiomyocytes were pretreated with or without LXA₄ or HO-1 specific interfering RNA (siRNA) or various blockers and openers of K⁺ channels before exposing to H/R injury. The levels of lactate dehydrogenase (LDH) and creatine kinase (CK) in cellular supernatants and necrosis factor-α (TNF-α) in cellular lysates were measured by using ELISA. Expressions of HO-1 mRNA and protein were analyzed by using RT-PCR and Western blot respectively. Pretreatment of the cells undergoing H/R injury with LXA₄ significantly reduced the LDH and CK levels induced by H/R injury, and increased the expressions and activity of HO-1. However, the protective effects of LXA₄ were completely blocked by transfection of the cells with HO-1 siRNA, and were partially but significantly blocked by pretreatment of the cells with various blockers of K⁺ channels. The LXA₄-induced expressions of HO-1 in the cells were also inhibited by HO-1 siRNA and various blockers of K⁺ channels. The inhibitory effects of LXA₄ on enhanced TNF-α levels induced by H/R injury were abolished by transfection of the cells with HO-1 siRNA. In conclusion, the protective role of LXA₄ on cardiomyocytes against H/R injury is related to upregulation of HO-1 via reduced production of TNF-α and activation of ATP-sensitive K⁺ channels and calcium-sensitive K⁺ channel.     

Protective effect of nicotinamide on neuronal cells under oxygen and glucose deprivation and hypoxia/reoxygenation

Nicotinamide (vitamin B₃) reduces the infarct volume following focal cerebral ischemia in rats; however, its mechanism of action has not been reported. After cerebral ischemia and/or reperfusion, reactive oxygen species (ROS) and reactive nitrogen species may be generated by inflammatory cells through several cellular pathways, which can lead to intracellular calcium influx and cell damage. Therefore, we investigated the mechanisms of action of nicotinamide in neuroprotection under conditions of hypoxia/reoxygenation. Results showed that nicotinamide significantly protected rat primary cortical cells from hypoxia by reducing lactate dehydrogenase release with 1 h of oxygen-glucose deprivation (OGD) stress. ROS production and calcium influx in neuronal cells during OGD were dose-dependently diminished by up to 10 mM nicotinamide (p<0.01). This effect was further examined with OGD/reoxygenation (H/R). Cells were stained with the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI) or antibodies against anti-microtubule-associated protein-2 and cleaved caspase-3. Apoptotic cells were studied using Western blotting of cytochrome c and cleaved caspase-3. Results showed that vitamin B₃ reduced cell injury, caspase-3 cleavage and nuclear condensation (DAPI staining) in neuronal cells under H/R. In addition, nicotinamide diminished c-fos andzif268 immediate-early gene expressions following OGD. Taken together, these results indicate that the neuroprotective effect of nicotinamide might occur through these mechanisms in this in vitro ischemia/reperfusion model.     

A pathway of neuronal apoptosis induced by hypoxia/reoxygenation: roles of nuclear factor-kappaB and Bcl-2

As a model of the reperfusion injury found in stroke, we have exposed neurons to hypoxia followed by reoxygenation. Neurons treated with hypoxia/reoxygenation (H/R) respond by activating nuclear factor-kappaB (NFkappaB), releasing cytochrome c from their mitochondria, and ultimately dying. Further supporting an apoptotic mechanism, expression of the antiapoptotic Bcl-2 and Bcl-x proteins was increased following H/R. In this model, adenoviral-mediated transduction of lkappaB expression inhibited NFkappaB activation and significantly accelerated cytochrome c release and caspase-dependent neuronal death. At the same time, expression of mutated lkappaB prevented the increased expression of endogenous Bcl-2 and Bcl-x. In the presence of mutated lkappaB, singular overexpression of only Bcl-2 by adenoviral-mediated transduction significantly inhibited cytochrome c release, caspase-3-like activation, and cell death in response to H/R. These findings suggest a pathway where NFkappaB activation induces overexpression of Bcl-2 and Bcl-x, which function to prevent apoptotic cell death following H/R treatments.     

Small interference RNA against PTP-1B reduces hypoxia/reoxygenation induced apoptosis of rat cardiomyocytes

Objective To investigate the effect of siRNA against PTP-1B on neonatal rat cardiac myocyte apoptosis induced by hypoxia-reoxygenation (H/R) and its molecular mechanisms. Methods Isolated neonatal and adult rat cardiac myocytes were cultured for 24 h after PTP-1B siRNA transfection, and with 2, 4 and 6 h of hypoxia followed by 6 h of reoxygenation (H/R). The cardiac myocyte apoptosis induced by the treatments was assessed by TUNEL staining. Levels of PTP-1B and phospho-Akt were determined by Western blot, colorimetric assay kits were used to measure activities of caspase-3 and 8, and co-immunoprecipitation was used to check the amount of PTP-1B bound to FasR. Sodium orthovanadate, a general pharmacological phosphatase blocker and LY294002, an inhibitor of PI3-kinase/Akt pathway, were respectively used to inhibit PTP-1B and Akt activity. Results H/R resulted in severe injury in cultured rat cardiomyocytes and upregulated PTP-1B expression. However, siRNA against PTP-1B significantly decreased the number of apoptotic cardiomyocytes induced by 4H/6R as compared with cells without siRNA treatment (Apoptotic index: 12.1 ± 1.4% vs. 23.2 ± 1.6%, P < 0.05), along with greater phosphorylation of Akt, reduced activities of caspase-3 and 8, and the lower association of PTP-1B with FasR. Vanadate and LY294002 also partly reduced apoptosis of cardiomyocytes induced by 4H/6R. Conclusions PTP-1B is a key regulator of apoptosis of cardiomyocytes induced by H/R, and siRNA against PTP-1B effectively protects cardiomyocytes against H/R injury, the mechanisms of which might be associated with Akt activation, the reduction of both caspase-3 and 8 activities, and the lower amount of PTP-1B bound to FasR.     

干扰FSCN1基因表达对前列腺癌细胞凋亡、活性氧水平影响的研究

目的探讨干扰FSCN1基因表达对前列腺癌细胞凋亡、活性氧(ROS)水平影响及机制。方法以正常前列腺上皮细胞RWPE-1为对照细胞,通过RT-PCR及Western blot检测前列腺癌LNCaP、DU145和PC-3细胞中FSCN1 mRNA及蛋白表达;以LipofectamineTM 2000为载体,DU145细胞分为si-FSCN1组(靶向抑制FSCN1的小干扰RNAs转染DU145细胞)、阴性对照组(随机序列转染DU145细胞)及空白对照组(未转染的细胞),siRNA转染48 h,Western blot检测FSCN1、PCNA、NF-κB p65、p-NF-κB p65、IKKα和p-IKKα的蛋白表达。CCK8检测细胞活力,流式细胞术检测细胞凋亡率及ROS水平。多组间比较采用单因素方差分析,组间两两比较采用SNK-q检验。结果与RWPE-1细胞比较,LNCaP、DU145和PC-3细胞中FSCN1 mRNA(1比2.561±0.189、7.183±0.882、4.796±0.567、4.796±0.567)及蛋白表达(0.053±0.007比0.217±0.013、0.654±0.058、0.316±0.035)均升高,差异具有统计学意义(P均<0.05)。与阴性对照组比较,si-FSCN1组FSCN1表达(0.473±0.052比0.086±0.010)降低,差异具有统计学意义(P均<0.05)。与si-FSCN1组比较,空白对照组、阴性对照组细胞活力(0.302±0.033比0.787±0.069、0.764±0.063)均升高,凋亡率(24.54﹪±1.47﹪比3.04﹪±0.36﹪、3.28﹪±0.40﹪)和ROS相对荧光强度(90.04±5.73比47.88±3.62、49.62±4.11)均降低,差异具有统计学意义(P均<0.05)。与si-FSCN1组比较,空白对照组、阴性对照组PCNA(0.255±0.032比0.654±0.062、0.631±0.058)、NF-κB p65(0.092±0.011比0.296±0.032、0.318±0.037)、p-NF-κB p65(0.042±0.008比0.155±0.018、0.151±0.016)、IKKα(0.112±0.01比0.172±0.020、0.192±0.023)和p-IKKα的蛋白表达(0.051±0.005比0.102±0.011、0.091±0.009)均升高,Cleaved caspase3蛋白表达(0.206±0.018比0.074±0.009、0.085±0.010)均降低,差异具有统计学意义(P均<0.05)。阴性对照组与空白对照组细胞活力、凋亡率、ROS水平及FSCN1、PCNA、Cleaved caspase3、NF-κB p65、p-NF-κB p65、IKKα和p-IKKα的蛋白表达差异均无统计学意义(P>0.05)。结论干扰FSCN1基因表达可降低前列腺癌细胞活力及诱导凋亡,机制可能与ROS水平升高及NF-κB信号下调有关。     

Silencing MEKK3 attenuates cardiomyocyte injury caused by hypoxia/reoxygenation via the sonic hedgehog pathway

MEKK3 is a member of MAP3K, which plays a pivotal role in cardiac diseases. In this study, we aimed to investigate the effects and potential mechanisms of MEKK3 on hypoxia/reoxygenation (H/R) injury of cardiomyocytes. After exposing H9C2 cells to H/R insult, real-time polymerase chain reaction and western blot analysis showed that MEKK3 was highly expressed. Cell viability, cell apoptosis, caspase 3/7 activity, and cleaved-caspase 3 expression were tested using a CCK-8 assay, Cell Death Detection PLUS ELISA, Caspase-Glo 3/7 Assay Kit and western blot analysis, respectively. Mitochondrial membrane potential, cytochrome C expression, adenosine triphosphate (ATP), and reactive oxygen species also were measured using JC-1 staining, western blot analysis, an ATP Assay Kit, and DCFH ₂-DA staining, respectively. The messenger RNA (mRNA) levels and secretions of TNF-α, IL-6, and IL-1β were evaluated. The results revealed that MEKK3 silencing promoted cell survival and attenuated lactate dehydrogenase leakage, cell apoptosis, caspase 3/7 activity, and the protein level of cleaved-caspase 3. Moreover, knockdown of MEKK3 blocked mitochondrial impairment by inhibiting the loss of mitochondrial membrane potential and cytochrome C expression as well as promoting ATP synthesis. MEKK3 deficiency led to a decrease in reactive oxygen species and malondialdehyde (MDA) generation and an increase in superoxide dismutase (SOD) activity. Deletion of MEKK3 led to reduced inflammatory cytokines in mRNA level and secretion. MEKK3 suppression activated the sonic hedgehog (Shh) signaling pathway in H9C2 cells. After blocking the Shh signaling pathway with a specific inhibitor, cyclopamine, the cardioprotective functions of MEKK3 downregulation were partly abolished. In conclusion, downregulation of MEKK3 prevented apoptosis and inflammation in H9C2 cells via the Shh signaling pathway.     

Mitochondrial involvement in IGF-1 induced protection of cardiomyocytes against hypoxia/reoxygenation injury

Studies in animal models of myocardial ischemia-reperfusion revealed that the administration of insulin-like growth factor (IGF-1) can provide substantial cardioprotective effect. However, the mechanisms by which IGF-1 prevents myocardial ischemia-reperfusion injury are not fully understood. This study addresses whether mitochondrial bioenergetic pathways are involved in the cardioprotective effects of IGF-1. Single cardiomyocytes from adult rats were incubated in the absence or presence of IGF-1 for 60 min and subjected to 60 min hypoxia followed by 30 min reoxygenation at 37°C. Mitochondrial function was evaluated by assessment of enzyme activities of oxidative phosphorylation and Krebs cycle pathways. Hypoxia/reoxygenation (HR) caused significant inhibition of mitochondrial respiratory complex IV and V activities and of the Krebs cycle enzyme citrate synthase, whereas pretreatment with IGF-1 maintained enzyme activities in myocytes at or near control levels. Mitochondrial membrane potential, evaluated with JC-1 staining, was significantly higher in IGF-1 + HR- treated myocytes than in HR alone, with levels similar to those found in normal control cardiomyocytes. In addition, IGF-1 reduced both HR-induced lactate dehydrogenase (LDH) release and malondialdehyde production (an indicator of lipid peroxidation) in cardiomyocytes. These results suggest that IGF-1 protects cardiomyocytes from HR injury via stabilizing mitochondria and reducing reactive oxidative species (ROS) damage.     

细胞粘附分子在中性粒细胞介导的缺氧／复氧心肌细胞损伤中的作用

目的观察缺氧/复氧对心肌细胞与中性粒细胞粘附效应的影响及细胞间粘附分子-1(intercellularadhensionmolecule-1,ICAM-1)和淋巴细胞功能相关抗原-1(lymphocytefunctionassociatedantigen-1,LFA-1)在中性粒细胞介导的心肌细胞损伤的作用。方法计数不同实验条件下与心肌细胞粘附的中性粒细胞;以及抗ICAM-1单抗和抗LFA-1单抗阻断后中性粒细胞粘附数的改变,检测心肌细胞乳酸脱氢酶释放量。结果中性粒细胞与缺氧/复氧心肌细胞粘附数较缺氧组和正常对照组显著增加(P＜0.01);心肌细胞释放LDH明显增高(P＜0．01),单纯缺氧组与正常对照组相比无显著差异(P＞0．05)。加入抗ICAM-1单抗和抗LFA-1单抗后,缺氧/复氧组与心肌细胞粘附的中性粒细胞数较正常对照组显著降低(P<0．01),心肌细胞释放LDH也明显下降(P＜0．01)。缺氧组与正常对照组相比则无显著差异(P＞0.05)。结论缺氧/复氧使心肌细胞与中性粒细胞粘附效应增加,心肌细胞损伤加重,ICAM-1和LFA-1参与这一过程。抗ICAM-1和抗LFA-1单抗可减轻中性粒细胞对缺氧/复氧心肌细胞的损伤。     

右美托咪定对A549细胞缺氧/复氧损伤时细胞凋亡及CHOP的影响

目的：探讨右美托咪定（Dex）对缺氧/复氧所致的A549细胞（起源于肺泡Ⅱ型上皮细胞系）损伤及对CCAAT/增强子结合蛋白同源蛋白（CHOP）表达的影响。方法：将处于对数生长期的A549细胞随机分为4组（n=10）：常氧培养组（N组）,Dex常氧组（D组）,缺氧/复氧组（H组）,缺氧/复氧+Dex组（HD组）。D组和HD组在造模开始时加入1 nmol/L Dex,N组和D组细胞常氧培养30 h,H组和HD组细胞缺氧6 h,复氧24 h。之后用倒置显微镜观察细胞形态学变化。采用CCK-8法检测A549细胞活力。原位末端标记（TUNEL）法检测A549细胞的凋亡指数（AI）。蛋白免疫印迹法（Western blot）和逆转录-聚合酶链反应（RT-PCR）分别检测A549细胞CHOP、Grp78、caspase-3蛋白和CHOP、Grp78 mRNA表达水平。结果：与N组比较,H组细胞数量减少,细胞形态发生改变。A549细胞的吸光度值明显下降（P<0.01）,AI值升高（P<0.01）,凋亡细胞数明显增加。CHOP、Grp78、caspase-3蛋白和CHOP、Grp78 mRNA表达显著上升（P<0.01）。与H组相比,HD组细胞损伤减轻,吸光度值上调（P<0.01）,凋亡细胞数明显减少（P<0.01）。CHOP、caspase-3蛋白,CHOP mRNA表达降低（P<0.01）。结论：Dex可有效减少缺氧/复氧引起的A549细胞凋亡,其机制可能与Dex对抗CHOP信号通路所致的凋亡有关。     

VDAC1 promotes cardiomyocyte autophagy in anoxia/reoxygenation injury via the PINK1/Parkin pathway

Ischemia/reperfusion (I/R) is a well-known injury to the myocardium, but the mechanism involved remains elusive. In addition to the well-accepted apoptosis theory, autophagy was recently found to be involved in the process, exerting a dual role as protection in ischemia and detriment in reperfusion. Activation of autophagy is mediated by mitochondrial permeability transition pore (MPTP) opening during reperfusion. In our previous study, we showed that MPTP opening is regulated by VDAC1, a channel protein located in the outer membrane of mitochondria. Thus, upregulation of VDAC1 expression is a possible trigger to cardiomyocyte autophagy via an unclear pathway. Here, we established an anoxia/reoxygenation (A/R) model in vitro to simulate the I/R process in vivo. At the end of A/R treatment, VDAC1, Beclin 1, and LC3-II/I were upregulated, and autophagic vacuoles were increased in cardiomyocytes, which showed a connection of VDAC1 and autophagy development. These variations also led to ROS burst, mitochondrial dysfunction, and aggravated apoptosis. Knockdown of VDAC1 by RNAi could alleviate the above-mentioned cellular damages. Additionally, the expression of PINK1 and Parkin was enhanced after A/R injury. Furthermore, Parkin was recruited to mitochondria from the cytosol, which suggested that the PINK1/Parkin autophagic pathway was activated during A/R. Nevertheless, the PINK1/Parkin pathway was effectively inhibited when VDAC1 was knocked-down. Taken together, the A/R-induced cardiomyocyte injury was mediated by VDAC1 upregulation, which led to cell autophagy via the PINK1/Parkin pathway, and finally aggravated apoptosis.     

NUMBL interacts with TRAF6 and promotes the degradation of TRAF6

Tumor necrosis factor-associated factor 6 (TRAF6) is an essential adaptor protein for IL-1R or TLR-mediated NF-κB signaling pathway activation. In previous work we have found NUMBL interacts with TAB2 and negatively regulates NF-κB signaling pathway. Here, we report that NUMBL directly binds to TRAF6 in vivo and in vitro. NUMBL down-regulates TRAF6 protein level and shortens its half-life. Furthermore, knockdown of NUMBL significantly increases endogenous TRAF6 protein level in the cultured cortical neurons. In vivo ubiquitination assays indicate that NUMBL promotes the assembly of K48-linked polyubiquitination chains on TRAF6, but has no significant effect on its K63-linked polyubiquitination. Our results collectively reveal that NUMBL interacts with TRAF6 and promotes the degradation of TRAF6 in vivo, leading to the inhibition of NF-κB signaling pathway.     

Signal transduction of MEK/ERK and PI3K/Akt activation by hypoxia/reoxygenation in renal epithelial cells

The extracellular signal-regulated kinase (ERK) and Akt have been reported to be activated by ischemia/reperfusion in vivo. However, the signaling pathways involved in activation of these kinases and their potential roles were not fully understood in the postischemic kidney. In the present study, we observed that these kinases are activated by hypoxia/reoxygenation (H/R), an in vitro model of ischemia/reperfusion, in opossum kidney (OK) cells and elucidated the signaling pathways of these kinases. ERK and Akt were transiently activated during the early phase of reoxygenation following 4-12h of hypoxia. The ERK activation was inhibited by U0126, a specific inhibitor of ERK upstream MAPK/ERK kinase (MEK), but not by LY294002, a specific inhibitor of phosphoinositide 3-kinase (PI3K), whereas Akt activation was blocked by LY294002, but not by U0126. Inhibitors of epidermal growth factor receptor (EGFR) (AG 1478), Ras and Raf, as well as antioxidants inhibited activation of ERK and Akt, while the Src inhibitor PP2 had no effect. PI3K/Akt activation was shown to be associated with up-regulation of X chromosome-linked inhibitor of apoptosis (XIAP), but not survivin. Reoxygenation following 4-h hypoxia-stimulated cell proliferation, which was dependent on ERK and Akt activation and was also inhibited by antioxidants and AG 1478. Taken together, these results suggest that H/R induces activation of MEK/ERK and PI3K/Akt/XIAP survival signaling pathways through the reactive oxygen species-dependent EGFR/Ras/Raf cascade. Activation of these kinases may be involved in the repair process during ischemia/reperfusion.     

The Ethanol Extract of Syringa oblata Heartwood,a Mongolian Folk Medicine Containing Major Lignans,Exerts Analgesic and Sedative Effects on Mice

The heartwood of Syringa oblata Lindl. (SO) is one of Mongolian folk medicines to treat insomnia and pain, while its pharmacological evaluation and underlying mechanism remain unclear. In this study, the sedative effect of ethanol extract of SO (ESO) was evaluated with the locomotor activity test and the threshold dose of pentobarbital sodium-induced sleep test in mice, and the hot plate test, acetic acid-induced writhing test, and formalin test in mice were used to evaluate its analgesic effect. The underlying mechanism of ESO analgesia was explored by RT-PCR and western blot analysis, which is associated with the regulation of the NF-κB signaling pathway. Besides, the main constituents of ESO were characterized by LC/MS data analysis and comparison with isolated pure compounds. The current findings brought evidence for clinical application and further pharmacological and phytochemical studies on SO.     

Short-term effects of anastrozole treatment on insulin-like growth factor system in postmenopausal advanced breast cancer patients

Insulin-like growth factors (IGFs) play a fundamental role in cancer development by acting in both an endocrinal and paracrinal manner, and hormone breast cancer treatments affect the IGF system by modifying circulating growth factor levels. We evaluated total IGF-1, IGF-2, IGF binding protein (IGFBP)-1 and IGFBP-3 in the blood of 34 postmenopausal advanced breast cancer patients (median age 63 years, range 41–85) treated with anastrozole, a non-steroidal structure aromatase inhibitor (NSS-AI). The plasma samples were obtained at baseline, and after 2, 4, 8 and 12 weeks of treatment. The IGFs were quantitated by means of sensitive radioimmunoassays (RIAs). IGF-1 significantly increased during anastrozole treatment (baseline versus 12 weeks, P=0.031), IGF-2 showed a trend towards an increase, and IGFBP-1 constantly but not significantly decreased; IGFBP-3 did not seem to be affected at all. The anastrozole-induced changes in IGFs and IGFBP-1 appeared to be different in the patients receiving a clinical benefit from those observed in non-responders. We have previously shown that letrozole (a different type of NSS-AI) modifies blood IGF-1 levels, and the results of this study of the biological effects of anastrozole on the components of the IGF system confirm our previous observations.     

毛喉鞘蕊花提取物对大鼠哮喘模型炎症介质的影响及机制研究

为探讨毛喉鞘蕊花提取物(Coleus Forskohlii extract,CFE)对哮喘的治疗作用及机制,采用卵清蛋白(ovalbumin,OVA)结合佐剂氢氧化铝建立大鼠哮喘模型,SPF级SD雄性大鼠60只,随机分为5组,即空白组、模型组、地塞米松组、CFE高剂量组(12.8g/kg)和CFE低剂量组(6.4g/kg),分别用生理盐水、地塞米松及CFE每天灌胃1次,连续14天,灌胃容积为1mL/100g。通过HE染色观察肺组织病理学形态学变化;ELISA法测定大鼠血清(serum)及肺泡灌洗液(BALF)中IFN-γ、IL-4和IL-17A含量变化;Western blot检测MAPK通路相关蛋白的表达变化。HE结果显示,与模型组相比,CFE高、低剂量组能显著减轻OVA刺激后大鼠肺组织病理性损伤,气道轮廓清晰,上皮细胞脱落明显减少;ELISA结果显示,与模型组相比,CFE高、低剂量组大鼠血清及BALF中IL-4、IL-17A的含量明显下降,IFN-γ的含量明显上升(P<0.05或P<0.01);WB结果显示,CFE高剂量组可显著降低大鼠肺组织p-ERK、p-JNK及p-p38蛋白表达(P<0.05或P<0.01)。综上,毛喉鞘蕊花提取物可有效缓解OVA诱导的大鼠哮喘气道炎症反应,其机制与调节MAPK信号传导通路有关。     

A20 overexpression exerts protective effects on podocyte injury in lupus nephritis by downregulating UCH-L1

Lupus nephritis (LN), an autoimmune kidney disease caused by systemic lupus erythematosus (SLE), is the inflammation of the kidney. Although the treatment of LN is still a therapeutic challenge for many practitioners, the present study aims to provide a new insight for the treatment and management. The study aims to explore the effect of A20 on LN in relation to the nuclear factor-kappa B (NF-κB) signaling pathway. MRL/lpr mice were used as the LN mouse model. Next, A20, UCH-L1, and NF-κB expression in LN patients and MRL/lpr mice was determined. A20 was upregulated in podocytes to assess biological functions of A20 in LN. Furthermore, to further investigate the pivotal role of the NF-κB pathway in LN, the NF-κB pathway was blocked in podocytes. Next, UCH-L1 was downregulated in MRL/lpr mice to assess biological functions of UCH-L1 in LN. A20 was downregulated, whereas UCH-L1 was upregulated in LN. Overexpressed A20 declined NF-κB, UCH-L1 expression, and the extent of p65 phosphorylation. A20 overexpression or UCH-L1 inhibition increased expression of synaptoporin and nephrin but decreased desmin expression and ubiquitin accumulation level in podocytes. Moreover, A20 overexpression or UCH-L1 inhibition increased the podocyte number but decreased protein level of cleaved caspase-3, podocyte lesion improvement, decreased foot process width, glomerulus basement membrane, and foot process fusion rate. In addition, urine protein, blood urea nitrogen, serum creatinine, and ds-DNA antibody levels decreased with elevated A20 or depleted UCH-L1. Collectively, it could be concluded that A20 protects against podocyte injury in LN via UCH-L1 by inactivating the NF-κB signaling pathway.