The involvement of p38 MAPK in transforming growth factor β1-induced apoptosis in murine hepatocytes

We reported in this manuscript that TGF-beta1 induces apoptosis in AML12 murine hepatocytes, which is associated with the activation of p38 MAPK signaling pathway. SB202190, a specific inhibitor of p38 MAPK, strongly inhibited the TGF-beta1-induced apoptosis and PAI-1 promoter activity. Treatment of cells with TGF-beta1 activates p38. Furthermore, over-expression of dominant negative mutant p38 also reduced the TGF-beta1-induced apoptosis. The data indicate that the activation of p38 is involved in TGF-beta1-mediated gene expression and apoptosis.

Overexpression of heme oxygenase-1 protects smooth muscle cells against oxidative injury and inhibits cell proliferation

To investigate whether the expression of exogenous heme oxygenase-1 (HO-1) gene within vascular smooth muscle cells (VSMC) could protect the cells from free radical attack and inhibit cell proliferation, we established an in vitro transfection of human HO-1 gene into rat VSMC mediated by a retroviral vector. The results showed that the profound expression of HO-1 protein as well as HO activity was 1.8- and 2.0-fold increased respectively in the transfected cells compared to the non-transfected ones. The treatment of VSMC with different concentrations of H2O2 led to the remarkable cell damage as indicated by survival rate and LDH leakage. However, the resistance of the HO-1 transfected VSMC against H2O2 was significantly raised. This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP-IX, a specific inhibitor of HO, for 24 h. In addition, we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of HO-1, and this effect might be related to decreased phosphorylation of MAPK. These results suggest that the overexpression of introduced hHO-1 is potentially able to reduce the risk factors of atherosclerosis, partially due to its cellular protection against oxidative injury and to its inhibitory effect on cellular proliferation.

Super-Resolution Image Reconstruction Based on an Improved Maximum a Posteriori Algorithm

A maximum a posteriori（ MAP） algorithm is proposed to improve the accuracy of super resolution（ SR） reconstruction in traditional methods. The algorithm applies both joints image registration and SR reconstruction in the framework,but separates them in the process of iteratiion. Firstly,we estimate the shifting parameters through two lowresolution（ LR） images and use the parameters to reconstruct initial HR images. Then,we update the shifting parameters using HR images. The aforementioned steps are repeated until the ideal HR images are obtained. The metrics such as PSNR and SSIM are used to fully evaluate the quality of the reconstructed image. Experimental results indicate that the proposed method can enhance image resolution efficiently.

Using Magnetic Method for the Identification of Anomalies Due to Kimberlite Pipes, Luando Area, Bié, Angola

One of the measurement geophysical methods to investigate kimberlite pipes is by using the magnetic method. The acquired field data in this study uses two proton-precession magnetometers for the mapping of magnetic anomalies due to kimberlites. Three different magnetic maps are obtained from the result of total magnetic field data processing on Oásis Montaj software programme. These maps include magnetic anomaly maps through statistical analyses, total magnetic field intensity map and map of the analytic signal. Based on the interpretation of these maps a structure is identified with SWW-NEE directions in which magnetic signatures that indicate the presence of kimberlite pipes are observed. As the interpretation of the magnetic anomalies is a complicated process due to their dipolar nature, the analytic signal is generated, where is possible to observe the typical shape of these anomalies.

Evodiamine induces extrinsic and intrinsic apoptosis of ovarian cancer cells via the mitogen-activated protein kinase/phosphatidylinositol-3-kinase/protein kinase B signaling pathways

OBJECTIVE: To explore the effects of evodiamine on ovarian cancer cells and the mechanisms underlying such effects.METHODS: Human ovarian cancer cells HO-8910 PM were treated with evodiamine at 0, 1.25,2.5, and 5 μM for 1-4 d. 3-(4,5-Dimethiylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was used to detect the growth inhibition rate of evodiamine-treated HO-8910 PM cells. The cell cycle was observed via propidium iodide(PI) staining. Apoptosis induction was assessed via Annexin V-fluorescein isothiocyanate/propidium iodide(Annexin V-FITC/PI) double staining assay. To verify the mechanism of apoptosis, caspase-dependent apoptotic pathway-related protein was detected by Western blot analysis. The expression levels of mitogen-activated protein kinase(MAPK)and/or phosphatidylinositol-3-kinase(PI3K)/pro-tein kinase B(Akt) pathway-related proteins were also investigated.RESULTS: Evodiamine significantly inhibited the proliferation of HO-8910 PM cells in a dose- and time-dependent manner. Evodiamine induced G2/M arrest with an increase of cyclin B1 level, and promoted cell apoptosis with a decrease of B cell lymphoma/lewkmia-2(Bcl-2) and an increase of Bcl-2-associated X protein(Bax) level. In addition,evodiamine treatment led to the activation of caspase-8, caspase-9, and caspase-3 and the cleavage of poly(ADP-ribose)-polymerase(PARP). Evodiamine targeted the MAPK and/or PI3K/Akt pathways by reducing the expression and activity of PI3 K, Akt, and extracellular signal-regulated kinase mitogen-activated protein kinase(ERK1/2 MAPK)and the activity of p38 MAPK.CONCLUSION: Evodiamine can inhibit the growth of ovarian cancer cells by G2/M arrest and intrinsic and extrinsic apoptosis. In addition, evodiamine-induced PI3K/Akt, ERK1/2 MAPK, and p38 MAPK signaling may be involved in cell death.

Busuishengxue ranules mediate their effects upon non-severe aplastic anemia via mitogen-activated protein kinase/extracellular signal-regulated kinase pathway

OBJECTIVE:To observe the clinical efficacy of Busuishengxue granules on non-severe aplastic anemia(NSAA)and investigate its effect on the mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK)pathway.METHODS:Sixty NSAA patients were divided equally into two groups.Subjects in the experimental group were treated with Busuishengxue granules,and the control group with Zaizaoshengxue tablets.The treatment course was 6 months and cu-rative efficacy was compared between the two groups as well as with 10 healthy individuals.Flow cytometry(FCM)was used to detect the intracellular concentration of Ca2+([Ca2+]i).Western blotting was employed to detect the expression of enzymes in the MAPK/ERK pathway.RESULTS:The efficacy of Busuishengxue granules was significantly better than that of Zaizaoshengxue tablets(P<0.05).Before treatment,expression of JNK,phospho-ERK 1/2 and p-JNK was higher,and[Ca2+]i higher,than that of the control group(P<0.05).After treatment with Busuishengxue granules,expression of all enzymes related to signal transduction pathways in the blood cells of NSSA patients were altered to different degrees.CONCLUSION:Busuishengxue granules had a better effect with regard to improving symptom scores,increasing the number of blood leukocytes,and increasing hemoglobin levels than Zaizaoshengxue tablets,and they differed slightly in terms of increasing the number of platelets.

Effect of Chaihushugan San on expression of the Raf/mitogen-activated protein kinase/extracellular signal-regulated kinase pathway in the hippocampi of perimenopausal rats induced by immobilization stress

OBJECTIVE: We wished to study the impact of Chaihushugan San(CSS) on the behavior of perimenopausal rats with liver-Qi stagnation(LQS) and to investigate the effect of CSS on signal transduction of the Raf/mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase(ERK) cascade in the hippocampi of rats induced by immobilization.METHODS: Twenty 52-week-old female rats were divided into two groups by the random number table method: model control group(MCG) and CSS group(CSSG), with 10 rats in each group.Ten-week-old female rats were used as the normal control group(NCG). CSS effects were assessed using rats exposed to immobilization stress by measuring body weight and sucrose consumption, serum hormone levels, and observing performance in the open field test(OFT). Molecular mechanisms were examined by measuring the effect of CSS on expression of Raf1, MEK1/2 and ERK1/2 m RNA in hippocampi using quantitative real-time polymerase chain reaction and by measuring levels of these proteins and related phospho-proteins using Western blotting.RESULTS: Perimenopausal rats with LQS had decreased locomotor activity; reduced sucrose consumption; and increased serum levels of corticotropin releasing hormone(CRH) and corticosterone(CORT). Activation of hippocampal Raf/MEK/ERK cascade was suppressed significantly in the MCG,and activation was increased after 21 days of CSS treatment.CONCLUSION: CSS has significant effects upon relief of the symptoms of LQS in immobilization-induced rats. The mechanism underlying this action might(at least in part) be mediated by reversal of disruption of the Raf/MEK/ERK pathway.

Effect of Zishenpingchan granule prepared from Chinese medicinal substances on the c-Jun N-terminal protein kinase pathway in mice with Parkinson's disease induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine

OBJECTIVE:To investigate the regulatory mechanism of the c-Jun N-terminal protein kinase(JNK)signaling pathway in substantia nigra(SN) dopaminergic neurons inflammation and apoptosis, and the neuroprotective effect of Zishenpingchan granules in mice with Parkinson's disease(PD) induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).METHODS:PD model mice were established by intraperitoneally injecting MPTP.Sixty mice were divided into a model group, Traditional Chinese Medicine(TCM) group and control group.The mice of the TCM group were administered Zishenpingchan granules 7 days before PD induction.Seven days after PD induction, we examined locomotor activity,and performed the rotarod test and swimming test,to evaluate limb movement function.Furthermore,we used immunohistochemistry and western blotting to examine the expression of tyrosine hydroxylase(TH), cyclooxygenase-2(Cox-2), caspase-3 and p-JNK.The terminal deoxynucleotidyl transferase mediated d UTP nick end labeling(TUNEL) method was used to examine neuron apoptosis in the SN.RESULTS:Compared with the control group, the mean score of locomotor activity, rotarod test and swimming test was significantly lower in the model group(P < 0.05); the TH-positive neuron expression was significantly decreased in the SN pars compacta(SNpc); the protein expression levels of Cox-2,caspase-3 and p-JNK was obviously increased; and the number of TUNEL-positive neurons in the SN was increased(P < 0.01).Compared with the model group, the mean score of neurobehavioral tests in the TCM group was obviously higher, the loss of TH-positive neurons ignificantly decreased, the protein expression levels of Cox-2, caspase-3 and p-JNK obviously decreased, and the number of TUNEL-positive neurons in the SN clearly decreased(P < 0.01).CONCLUSION:The JNK pathway plays an important role in the regulation of inflammation and apoptosis in nigral cells in PD mice.TCM can suppress the over-activation of the JNK pathway in the SN, and alleviate the inflammatory response in nigral cells and dopaminergic neuron apoptosis in PD mice.

Efficacy of active compounds of Chanqin granules on airway neurogenic inflammation induced by PM2.5 in vivo

OBJECTIVE:To investigate the efficacy of active compounds of Chanqin(CQ)granules on PM2.5-induced airway neurogenic inflammation in vivo,and to elucidate the underlying mechanisms of action.METHODS:The Traditional Chinese Medicine systems pharmacology(TCMSP)database was searched,and the results were combined with oral bioavailability and drug analysis to identify the compounds in CQ granules.The pharmacophore modeling approach was used to predict the compound targets,and the diseases corresponding to the targets were obtained by searching the therapeutic target database(TTD),pharmacogenomics knowledgebase(Pharm GKB)and Drug Bank databases.Cytoscape software was used to construct the network pharmacological charts for Component-Target and Target-Disease interactions of the CQ granules.Then,the mechanisms of action and effectiveness of CQ granules for the treatment of PM2.5-induced airway neurogenic inflammation were analyzed.RESULTS:A total of 195 compounds and 171 targets were obtained from the analyses.A total of569 corresponding diseases were identified for these targets.Component-target and target-disease networks were constructed.The possible mechanisms and effective components in CQ granules for treating airway neurogenic inflammation were analyzed.Quercetin,kaempferol and isorhamnetin,beta-sitosterol and sitosterol,which are typically found in the formulation,have extensive pharmacological activities,including anti-inflammatory,antioxidant and antiviral actions and neuroprotective properties.Among these targets,androgen receptor,estrogen receptor,prostaglandin G/H synthase 2,and inducible nitric oxide synthase play important pathological roles,including the induction of neurogenic inflammation.CQ granules may have therapeutic effectiveness for numerous diseases in addition to respiratory diseases,including neoplasms,digestive system diseases,cardiovascular diseases,respiratory tract diseases and nervous system diseases.In vivo,CQ granules are effective in treating pulmonary inflammation and downregulate neuropeptides in the bronchoalveolar lavage fluid after PM2.5 exposure.CQ granules significantly decreased the levels of neurokinin A,neurokinin B and calcitonin gene-related peptide in the lung and dorsal root ganglia.CQ also significantly suppressed the upregulation of p-extracellular regulated protein kinase 1/2 and p-methyl ethyl ketone 1/2 induced by PM2.5 exposure.CONCLUSION:CQ granules have potential for thetreatment of neurogenic inflammation induced by PM2.5 in vivo,and the mechanism might involve downregulation of neuropeptides in the BALF,lung and dorsal root ganglia.

Sildenafil potentiates the proliferative effect of porcine pulmonary artery smooth muscle cells induced by serotonin in vitro

Background Sildenafil is one of the selective phosphodiesterase 5 inhibitors that has been proven by many investigators to suppress growth factor stimulated （e.g. platelet-derived growth factor （PDGF） or epidermal growth factor （EGF）） proliferation and hypertrophy of pulmonary artery smooth muscle cells （PASMCs） via cGMP/cGKla pathway. Serotonin promotes cell cycle progression leading to cell mitogenesis and plays a key role in the pathogenesis of pulmonary artery hypertension. The role of sildenafil in proliferation of PASMCs induced by serotonin has not been investigated so far. In this study we explored the underlying mechanism of the effect of sildenafil on serotonin induced proliferation of porcine PASMCs. Methods PASMCs were cells from primary cultures by the explant method from the pulmonary artery of swine and cells at passage 3-5 were used in this study. MTT colorimetric assay and flow cytometry analysis were used to evaluate the cell proliferation and alterations in cell cycle progression respectively. Western blotting analysis was applied to determine the expression of phosphorylated extracellular signal-regulated kinase （ERK）, proliferating cell nuclear antigen （PCNA） and mitogen activated protein kinase （MAPK） phosphatase-1 （MKP-1）. Results Serotonin （10 IJmol/L） induced the upregulation of phosphorylation of ERK1/ERK2 and PCNA, an increase in the percentage of cells in S phase and subsequent cell proliferation. Pretreatment with 1 μmol/L sildenafil potentiated the phosphorylation of ERK1/ERK2, an increase in the percentage of cells in S phase and cell proliferation, compared with serotonin stimulation alone （P〈0.05）. Furthermore, 30-minute pretreatment with 10 μmol/L U0126, specific antagonist for ERK kinase （MEK） prevented the increase in phosphorylation of ERK1/ERK2 and abolished cell cycle progression and the proliferation of PASMCs induced by sildenafil. Conclusion This study shows that sildenafil potentiated the proliferative effect of serotonin on PASMCs via phosphorylation of ERK1/ERK2.

Transcriptomic landscape regulated by the 14 types of bone morphogenetic proteins(BMPs)in lineage commitment and differentiation of mesenchymal stem cells(MSCs)

Mesenchymal stem cells(MSCs)are ubiquitously-existing multipotent progenitors that can self-renew and differentiate into multiple lineages including osteocytes,chondrocytes,adipocytes,tenocytes and myocytes.MSCs represent one of the most commonly-used adult progenitors and serve as excellent progenitor cell models for investigating lineagespecific differentiation regulated by various cellular signaling pathways,such as bone morphogenetic proteins(BMPs).As members of TGFb superfamily,BMPs play diverse and important roles in development and adult tissues.At least 14 BMPs have been identified in mammals.Different BMPs exert distinct but overlapping biological functions.Through a comprehensive analysis of 14 BMPs in MSCs,we demonstrated that BMP9 is one of the most potent BMPs in inducing osteogenic differentiation of MSCs.Nonetheless,a global mechanistic view of BMP signaling in regulating the proliferation and differentiation of MSCs remains to be fully elucidated.Here,we conducted a comprehensive transcriptomic profiling in the MSCs stimulated by 14 types of BMPs.Hierarchical clustering analysis classifies 14 BMPs into three subclusters:an osteo/chondrogenic/adipogenic cluster,a tenogenic cluster,and BMP3 cluster.We also demonstrate that six BMPs(e.g.,BMP2,BMP3,BMP4,BMP7,BMP8,and BMP9)can induce ISmads effectively,while BMP2,BMP3,BMP4,BMP7,and BMP11 up-regulate Smad-independent MAP kinase pathway.Furthermore,we show that many BMPs can upregulate the expression of the signal mediators of Wnt,Notch and PI3K/AKT/mTOR pathways.While the reported transcriptomic changes need to be further validated,our expression profiling represents the first-of-its-kind to interrogate a comprehensive transcriptomic landscape regulated by the 14 types of BMPs in MSCs.

Dehydroepiandrosterone indirectly inhibits human osteoclastic resorption via activating osteoblastic viability by the MAPK pathway

Background Dehydroepiandrosterone （DHEA） is widely known for its beneficial effect on postmenopausal osteoporosis, although the underlying mechanisms remain mainly unclear. In this study, we tried to determine the activation of mitogen-activated protein kinase signal pathways during DHEA treatment and the indirect role of osteoblasts （OBs） on osteoclasts under the DHEA treatment of postmenopausal osteoporosis. Methods Primary human OBs and osteoclast-like cells were cultured and, we pretreated OBs with or without U0126 （a highly selective inhibitor of both MEK1 and MEK2）. The OBs were treated with DHEA. We then tested the effects of DHEA on human osteoblastic viability, osteoprotegerin production and the expression of phosphor-ERK1/2 （extracellular signal-regulated kinase）. In the presence or absence of OBs, the function of osteoclastic resorption upon DHEA treatment was calculated. Results DHEA promoted the human osteoblastic proliferation and inhibited the osteoblastic apoptosis within the concentration range of 108-106 mol/L （P 〈0.05, P 〈0.01, respectively）. Within the effective concentration range, the expression of phosphor-ERK1/2 and osteoprotegerin was increased by DHEA and blocked by U0126. In the presence of OBs, DHEA could significantly decrease the number and the area of bone resorption lacuna （P 〈0.05 and P 〈0.01, respectively）. Without OBs, however, the effects of DHEA on the bone resorption lacuna were almost completely abolished. Conclusions DHEA could indirectly inhibit the human osteoclastic resorption through promoting the osteoblastic viability and osteoprotegerin production, which is mediated by mitogen-activated protein kinases signal pathway involving the phosphor-ERK1/2.