ABSTRACT
BACKGROUND: Activin B has been reported to promote the proliferation and migration of keratinocytes in vitro via the RhoA-JNK signaling pathway, whereas its in vivo role and mechanism in wound healing process has not yet been elucidated. PRINCIPAL FINDINGS: In this study, we explored the potential mechanism by which activin B induces epithelial wound healing in mice. Recombinant lentiviral plasmids, with RhoA (N19) and RhoA (L63) were used to infect wounded KM mice. The wound healing process was monitored after different treatments. Activin B-induced cell proliferation on the wounded skin was visualized by electron microscopy and analyzed by 5'-bromodeoxyuridine (BrdU) incorporation assay. Protein expression of p-JNK or p-cJun was determined by immunohistochemical staining and immunoblotting analysis. Activin B efficiently stimulated the proliferation of keratinocytes and hair follicle cells at the wound area and promoted wound closure. RhoA positively regulated activin B-induced wound healing by up-regulating the expression of p-JNK and p-cJun. Moreover, suppression of RhoA activation delayed activin B-induced wound healing, while JNK inhibition recapitulated phenotypes of RhoA inhibition on wound healing. CONCLUSION: These results demonstrate that activin B promotes epithelial wound closure in vivo through the RhoA-Rock-JNK-cJun signaling pathway, providing novel insight into the essential role of activin B in the therapy of wound repair.
Subject(s)
Activins/metabolism , Cell Proliferation , Epithelial Cells/metabolism , JNK Mitogen-Activated Protein Kinases/metabolism , Wound Healing/physiology , rhoA GTP-Binding Protein/metabolism , Animals , Apoptosis , Blotting, Western , Cell Movement , Cells, Cultured , Female , Hair Follicle/cytology , Hair Follicle/metabolism , Immunoenzyme Techniques , Keratinocytes/cytology , Keratinocytes/metabolism , Lentivirus , MAP Kinase Signaling System , Male , Mice , Phosphorylation , Signal Transduction , rho-Associated Kinases/metabolismABSTRACT
OBJECTIVE: To analyse and compare the chemical compositions in the volatile oil from Utramicro-powder and Common Grinding Powder of Cinnamomum. METHODS: The volatile oil was extracted by steam-stilling and analyzed by GC-MS. The relative content of each component was calculated by area normalization method. RESULTS: 34 and 19 peaks were isolated from Ultramicro-powder and Common Grinding Powder of Cinnamomum respectively. All of them were identified. CONCLUSION: The chemical components of the volatile oil from Ultramicro-powder and Common Grinding Powder of Cinnamomum are reported, the results here provides scientific proof for the application in external preparation of Cinnamomum.
Subject(s)
Acrolein/analogs & derivatives , Cinnamomum/chemistry , Oils, Volatile/chemistry , Plants, Medicinal/chemistry , Thymol/analysis , Acrolein/analysis , Acrolein/chemistry , Cymenes , Gas Chromatography-Mass Spectrometry , Monoterpenes/analysis , Monoterpenes/chemistry , Oils, Volatile/analysis , Oils, Volatile/isolation & purification , Particle Size , Plant Bark/chemistry , Powders , Thymol/chemistryABSTRACT
BACKGROUND: Cocaine addiction may involve complex neuroadaptations, including many changes of genes expression. Dopamine D3 receptors play an important role in cocaine addiction; however, its role in cocaine induced gene expression change is poorly understood. To identify the changes in gene expression induced by repeated cocaine exposure through D3 dopamine receptors, we compared the expression of four molecules: Janus kinase 2 (Jak2), g-aminobutanoic acid receptor subunit alpha 1 (GABAAalpha1), glutamate receptor AMPA3 alpha 3 (GluR 3) and stromal cell derived factor 1 (SDF1). These four have been implicated in mediating the actions of cocaine in the nucleus accumbens (NAc) and caudoputamen (CPu) in mice after acute and repeated cocaine exposure. METHODS: For the acute and repeated injections, the mice were divided into four groups: 30 mg/kg cocaine, nafadotride 0.5 mg/kg + cocaine 30 mg/kg, nafadotride 0.5 mg/kg, and saline as the basal group. The expression of Jak2, GABAAalpha1, GluR 3 and SDF1 were assayed by Western blot, quantitative real-time RT-PCR and immunohistochemistry. RESULTS: Twenty-four hours after seven consecutive days of repeated cocaine exposure, the expression of GABAAalpha1 decreased in cocaine group compared with basal line and further decreased in the cocaine + nafadotride group and remained at basal level in the nafadotride group. Similarly, the Jak2 expression decreased in cocaine group compared with base line. However, the levels of Jak2 increased in cocaine + nafadotride group compared with cocaine group, while remained at basal level in nafadotride group. CONCLUSIONS: GABAAalpha1 and Jak2 may be involved in chronic cocaine induced neuroadaptations. D3 dopamine receptors play an important role in the expression of these genes.
Subject(s)
Brain/drug effects , Cocaine/pharmacology , Janus Kinase 2/genetics , Receptors, Dopamine D3/physiology , Receptors, GABA-A/genetics , Animals , Brain/metabolism , Female , Gene Expression Regulation/drug effects , Immunohistochemistry , Janus Kinase 2/analysis , Male , Mice , Receptors, GABA-A/analysis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
OBJECTIVE: To study the role of dopamine receptors in the regulation of the activity of transcription factor cAMP response element-binding protein (CREB) after cocaine treatment. METHODS: By using dopamine receptor antagonists SCH23390 and nafadotride, the activation of CREB by D1 and D3 dopamine receptors after cocaine treatment and role of extracellular signal-regulated kinase (ERK) in cocaine-induced CREB activation were examined by Western blotting, which was also employed for determination of the effect of SCH23390 and nafadotride on CREB activation. RESULTS: D1 receptor antagonist could inhibit cocaine-induced CREB activation, while D3 receptor antagonist enhanced cocaine-induced CREB activation. Dopamine receptor antagonists SCH23390 and nafadotride did not induce CREB activation. SL327, a MEK inhibitor, inhibited cocaine-induced CREB activation. CONCLUSION: D1 and D3 dopamine receptors can oppositely regulate CREB activation after cocaine treatment and this regulation depends on ERK signaling pathway.
Subject(s)
Cocaine/pharmacology , Cyclic AMP Response Element-Binding Protein/metabolism , Receptors, Dopamine D1/physiology , Receptors, Dopamine D3/physiology , Animals , Benzazepines/pharmacology , Blotting, Western , Dopamine Antagonists/pharmacology , Dopamine Uptake Inhibitors/pharmacology , Extracellular Signal-Regulated MAP Kinases/metabolism , Mice , Naphthalenes/pharmacology , Pyrrolidines/pharmacology , Receptors, Dopamine D1/antagonists & inhibitors , Receptors, Dopamine D3/antagonists & inhibitors , Signal Transduction/drug effectsABSTRACT
Interleukin-2 (IL-2) can stimulate T cell proliferation and differentiation when binding to its receptor on T cells. It produces a marked effect by enhancing the cytotoxicity of CD8+ T cells and natural killer cells. Granulocyte-macrophage colony stimulating factor (GM-CSF) is associated with many cells proliferation, such as dendritic cells, macrophages. Here, we report the construction, expression and purification of a bifunctional protein, hIL-2/GM-CSF, which may facilitate interaction between T cells and the antigen presentation cells and improve the efficiency of antigen presentation. We found that the use of chemicals and temperature shift is a peculiar system for induction of the Escherichia coli transformed with an IPTG-regulated hIL-2/GM-CSF expression vector in this research. After renaturation, anion exchange chromatography, metal affinity chromatography, and strict endotoxin-free cation exchange chromatography, the fusion protein devoid of endotoxin showed high purity. Cell proliferation experiments proved that this bifunctional protein retains both hIL-2 and GM-CSF biological activities. These results will facilitate the numerous subsequent studies on this bifunctional molecule.
