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.

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.

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.