Endogenous Endothelin-1 Regulates Hypoxia-Induced Atrial Natriuretic Peptide Secretion by Activating the MAPK/ERK and PI3K/Akt Signaling Pathways in Isolated Beating Rabbit Atria

The present study investigated a possible mechanism for endogenous endothelin-1 (ET-1) regulation of atrial natriuretic peptide (ANP) secretion in isolated perfused acute hypoxic rabbit atria. Acute hypoxia significantly enhanced the release of ET-1 and the expression of the ET receptor (ETR) type A and B (ETRA and ETRB) in atrial tissues, with a concomitant increase in ANP secretion. The ETRA or ETRB antagonist, BQ123 (0.3 μmol/L) or BQ788 (0.3 μmol/L), respectively attenuated hypoxia-induced ANP secretion. Both antagonists significantly attenuated the levels of hypoxiainduced atrial phosphorylated (p)-extracellular signal-regulated kinase (ERK) and p-protein kinase B (Akt). The ERK and Akt inhibitors, PD098059 (30 μmol/L) and LY294002 (30 μmol/L), respectively mimicked the effect of the ETR antagonists. These results demonstrated that acute hypoxia- mediated atrial ET-1 regulated ANP secretion through ETR and the subsequent mitogenactivated protein kinase (MAPK)/ERK and ETR-phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathways. These pathways may mediate atrial endocrine functions under hypoxic conditions.

Arabidopsis NIMIN1,2 mediate redox homeostasis and JA/ET pathways to modulate Botrytis cinerea resistance

Arabidopsis NIM1-INTERACTING1 (NIMIN1) and NIMIN2 are required for salicylic acid (SA)-dependent resistance against biotrophic pathogens. In this study, we have demonstrated that NIMIN1, 2 are also essential for plant defense response to necrotrophic pathogen Botrytis cinerea. The nimin1 and nimin2 mutants displayed a higher susceptibility against B. cinerea than the wild type, which correlated with a decrease in B. cinerea-induced PDF).2 expression. Mutation in NIMIN1 or NIMIN2 enhanced accumulation of hydrogen peroxide (H_2O_2) with reductions in the activities of three main antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) at the early time (24h) of upon B. cinerea infection. These mutations also resulted in a visible decrease in B.cinerea-induced Ethylene Responsive Factor 1 (ERF1), Octadecanoid-Responsive Arabidopsis AP2/ERF 59 (ORA59), Allene Oxide Cyclase (AOC3), Opda Reductase 3 (OPR3),ACC Synthesis 2 (ACS2) and ACS6 expression, but an advance in MYC2 expression,indicating that NIMIN1,2 are essential for B. cinerea-activated jasmonic acid (JA)/ethylene(ET) biosynthesis and signaling. However, mutation in NIMIN1 or NIMIN2 drastically suppressed JA-, but not ET-activated PDF1.2 expression. Together, these results suggest that NIMIN1,2 may positively control the B. cinerea resistance by mediating redox homeostasis and JA/ET pathways in Arabidopsis.

Blockage of ETS homologous factor inhibits the proliferation and invasion of gastric cancer cells through the c-Met pathway

BACKGROUND Gastric cancer(GC)is one of the most common and deadliest types of cancer worldwide due to its delayed diagnosis and high metastatic frequency,but its exact pathogenesis has not been fully elucidated.ETS homologous factor(EHF)is an important member of the ETS family and contributes to the pathogenesis of multiple malignant tumors.To date,whether EHF participates in the development of GC via the c-Met signaling pathway remains unclear.AIM To investigate the role and mechanism of EHF in the occurrence and development of GC.METHODS The expression of EHF mRNA in GC tissues and cell lines was measured by quantitative PCR.Western blotting was performed to determine the protein expression of EHF,c-Met,and its downstream signal molecules.The EHF expression in GC tissues was further detected by immunohistochemical staining.To investigate the role of EHF in GC oncogenesis,small interfering RNA(siRNA)against EHF was transfected into GC cells.The cell proliferation of GC cells was determined by Cell Counting Kit-8 and colony formation assays.Flow cytometry was performed following Annexin V/propidium iodide(PI)to identify apoptotic cells and PI staining to analyze the cell cycle.Cell migration and invasion were assessed by transwell assays.RESULTS The data showed that EHF was upregulated in GC tissues and cell lines in which increased expression of c-Met was also observed.Silencing of EHF by siRNA reduced the proliferation of GC cells.Inhibition of EHF induced significant apoptosis and cell cycle arrest in GC cells.Cell migration and invasion were significantly inhibited.EHF silencing led to c-Met downregulation and further blocked the Ras/c-Raf/extracellular signal-related kinase 1/2(Erk1/2)pathway.Additionally,phosphatase and tensin homolog was upregulated and glycogen synthase kinase 3 beta was deactivated.Moreover,inactivation of signal transducer and activator of transcription 3 was detected following EHF inhibition,leading to inhibition of the epithelial-to-mesenchymal transition(EMT).CONCLUSION These results suggest that EHF plays a key role in cell proliferation,invasion,apoptosis,the cell cycle and EMT via the c-Met pathway.Therefore,EHF may serve as an antineoplastic target for the diagnosis and treatment of GC.

Mechanism of Inhibitory Effect of Essential Oil from Valerianae Jatamansi Rhizoma et Radix on Activation of Microglia

[Objectives]To investigate the inhibitory effect and possible mechanism of essential oil from Valerianae Jatamansi Rhizoma et Radix on microglia activation induced by lipopolysaccharide(LPS).[Methods]The LPS-induced microglia activation model was established and treated with different doses of essential oil from Valerianae Jatamansi Rhizoma et Radix.MTT assay was used to detect cell viability,ELISA to detect IL-6 secretion,RT-PCR to detect mRNA expression levels of IL-6,IL-1β,NF-κB,and IκBα,Western blotting to detect the protein expression of IL-6,IL-1β,NF-κB,IκBα,and their phosphorylated products.[Results]Compared with the normal control group,the model group showed increased IL-6 content(P<0.01),upregulated mRNA and protein levels of IL-6,IL-1β,and NF-κB(P<0.01),and elevated ratio of P-IκBα/IκBα(P<0.05).Compared with the model group,4 and 2μg/L essential oil from Valerianae Jatamansi Rhizoma et Radix reduced the content of IL-6(P<0.05),while 4,2,and 1μg/L essential oil from Valerianae Jatamansi Rhizoma et Radix down-regulated the mRNA and protein levels of IL-6,IL-1β,and NF-κB to varying degrees(P<0.05 or P<0.01),up-regulate the mRNA expression of IκBα(P<0.05 or P<0.01),and decreased the ratio of P-IκBα/IκBα(P<0.05 or P<0.01).[Conclusions]Essential oil from Valerianae Jatamansi Rhizoma et Radix can inhibit LPS-induced microglia activation,and its mechanism may be related to the inhibition of the NF-κB/IκBαsignaling pathway.

Study on the mechanism and active components of Radix et Rhizoma Rhei in the treatment of Alzheimer's disease based on molecular docking

Objective:To explore the mechanism and active components of Radix et Rhizoma Rhei in the treatment of Alzheimer's disease(AD)based on molecular docking.Methods:22 major components of Radix et Rhizoma Rhei were screened from TCMSP and related literatures,which docked with the key targets of NLRP3/Caspase-1/GSDMD signaling pathway.NLRP3,Caspase-1,GSDMD inhibitors MCC950,ML132 and LDC7559 were used as positive control to analyze the docking results.Results:The docking results showed that the main components of Radix et Rhizoma Rhei had different degrees of binding with NLRP3,Caspase-1 and GSDMD targets,and the potential active components were mutanochrome and physciondiglucoside.Conclusion:Molecular docking predicts that the main components of Radix et Rhizoma Rhei may act on NLRP3/Caspase-1/GSDMD signaling pathway,and the active components may be mutanochrome and physciondiglucoside,which provides theoretical basis for revealing the anti-inflammatory mechanism and active components of Radix et Rhizoma Rhei in the treatment of AD.

受壳寡糖诱导的油菜MAPK基因的克隆与分析

利用油菜cDNA芯片差异显示得到一个可被壳寡糖诱导的,与已知植物丝裂原活化蛋白激酶(MAPK)同源性高达94%的cDNA片段,利用cDNA末端快速扩增(RACE)的方法得到了该片段全长,与拟南芥中AtMPK4具有高同源性,故将其命名为BnMPK4登录到NCBI Gen Bank(DQ206628)。用生物信息学方法分析,该基因属于植物MAPK的B家族,具有植物MAPK典型的TXY保守区域,在C端也含有一个在B家族中保守的CD区。利用半定量RT-PCR检测发现BnMPK4在叶片中表达且可被茉莉酸(JA)和壳寡糖诱导,但对水杨酸(SA)和脱落酸(ABA)响应不明显。推测该基因在壳寡糖诱抗信号转导和JA/ET信号通路中起关键作用。

本生烟感染TNV后ET/JA-依赖的防御信号的初步研究

本生烟(Nicotiana benthamiana)与烟草坏死病毒大豆分离物(TNV-A^C)构成的植病互作体系是一种特殊的亲和互作.向本生烟外施一定剂量的抗性诱导剂茉莉酸(JA)、乙烯(ET),和抑制剂丙基没食子酸(PG,JA合成抑制剂)、氯化钴(CoCl_2,ET合成阻断剂)后,用Northern blot,Western blot,半定量RT-PCR等方法,在病毒接种的早、中、晚期,检测TNV及防御基因PR1,PR3,GST1的表达变化.结果显示在诱导剂处理后,TNV表达延迟,病毒含量降低,而抑制剂的作用则相反.防御基因的变化与病毒复制量的变化趋势相同,以PR3最为显著.结果初步表明ET/JA是本生烟-TNV互作系统防御信号途径的关键分子.

茉莉酸和乙烯信号转导途径参与花生应答果腐病病原菌侵染的转录组测序分析

花生果腐病是花生生产上的一种重要病害,对花生的产量和品质构成了严重威胁。本研究以花育20号感病前后籽仁为样品,进行转录组测序,得到Unigene 60756个,其中表达量上调的基因10147个,下调的基因4334个;GO富集分析表明,差异表达基因被分类到24种细胞组分、5种分子功能和23种生物过程中;KEGG富集分析发现有4291个差异基因被注释到34条代谢通路中。筛选得到多条花生响应果腐病相关的激素信号转导途径,通过分析相关基因的表达图谱,推测花生可能通过JA/ET介导的抗病途径来抵抗果腐病病原菌的侵染。

6个玉米ERF基因的克隆、序列及表达分析

利用同源克隆的方法分离了6个玉米ERF(ethylene-responsive element binding factor)基因,分别命名为ZmERF4、ZmERF26、ZmERF28、ZmERF64、ZmERF89和ZmERF202。基因结构分析显示,ZmERF与拟南芥的AtERF可能存在差异。系统发育分析显示,ZmERFs与OsERFs的亲缘关系更加亲近,而与AtERFs的关系则稍远。RT-PCR表达分析表明,在乙烯利(ET)处理之后,这6个ZmERFs的mRNAs水平在叶中都是先积累后下降的,但其中4个基因(ZmERF4、ZmERF28、ZmERF64和ZmERF89)在玉米根中都是呈现负调控的。在脱落酸(ABA)和茉莉酸(JA)处理下,6个ZmERFs的mRNAs水平在叶中都发生了或多或少的变化,但是大部分基因在根中的表达量没有明显变化,特别是在JA处理之后。这些结果暗示ZmERFs可能在玉米根和叶的激素信号通路中发挥不同的作用。

Effect of herbal cake-partitioned moxibustion on Leptin/JAK#STAT3 in lipid-lowering pathway of hyperlipidemia rabbits

Objective:To observe the lipid-lowering effect of different transdermal absorption enhancers applied to the herbal cake-partitioned moxibustion in hyperlipidemia model rabbits,and to explore the possible mechanism.Methods:Forty New-Zea I a nd rabbits were ran domly divided into 5 groups using the ran dom nu mber table method,with 8 rats in each group.Rabbits in the blank group were fed routinely with normal diet;rabbits in the other groups were fed with high-fat diet for 12 weeks to establish the hyperlipidemia model.Rabbits in the blank and the model groups were not treated.After the model was prepared,rabbits in the non-transdermal absorption enhancer group received herbal cake-partitioned moxibustion without transdermal absorption enhancer;rabbits in the laurocapram group and the borneol group received herbal cake-partitioned moxibustion with laurocapram or borneol respectively.After 4 weeks of treatment,serum was collected for enzyme-linked immunosorbent assay(ELISA),and the liver tissues were isolated for imm uno histochemistry,qua ntitative polymerase chain reactio n(qPCR)and Western-blotting(WB)detecti on.Results:Serum ELISA results showed that leptin was significantly decreased in the model group compared with the blank group(P<0.05);compared with the model group,lepti n was significa ntly in creased in the non-tran sdermal absorpti on enhanee。the laurocapram and the borneol groups(all P<0.05);compared with the non-transdermal absorption enhancer group,leptin was significantly increased in the laurocapram group and the borneol group(both P<0.05);there was no significant differenee in leptin between the laurocapram and the borneol groups(P>0.05).The qPCR results of rabbit liver tissues showed that the mRNA expressions of leptin,Janus kinase 2(JAK2)and signal transducer and activator of transcription 3(STOT3)in the model group were significantly lower than those in the blank group(all P<0.05);compared with the model group,the mRNA expressions of leptin,leptin receptor(LR),JAK2 and S1AT3 in the non-transdermal absorptio n enhan cer,the laurocapram and the born eol groups were significantly in creased(all P<0.05);compared with the non-transdermal absorption enhancer group,the mRNA expressions of leptin,LR,JAK2 and S77VT3 in the laurocapram and the bor neol groups were sign ificantly in creased(all P<0.05);compared with the laurocapram group,the mRNA expressi ons of lepti n,LR,JAK2 and SW3 in the bor neol group were significa ntly in creased(P<0.05).The trend of immun ohistochemistry and WB detecti on results was basically con siste nt with the qPCR assay results.The immuno histochemistry and WB detection results of phosphorylated JAK2(phospho-JAK2)and phosphorylated S7AT3(phospho-STAT3)were basically consistent with those of JAK2 and S7AT3.Conclusion:The molecular expression of Leptin/JAK"S7AT3 pathway in the hyperlipidemia model rabbits was decreased.The molecular expression of Leptin/JAK0STCT3 pathway was significantly increased after the herbal cake-partitioned moxibustion.The application of laurocapram and borneol,as transdermal absorption enhancers,in the herbal cake-partitioned moxibustion could more obviously up-regulate the factors of the Leptin/JAK^SIAT3 lipid-regulating pathway than the herbal cake-partitioned moxibustion alone.