OBJECTIVE: To investigate the effect and possible mechanism of BDNF-AS on renal tubular epithelial cell injury induced by high glucose. METHODS: Human renal tubular epithelial cells HK-2 were cultured in vitro and transfected with BDNF-AS small interfering RNA or miR-145-5p mimic, or co-transfected with BDNF-AS small interfering RNA and miR-145-5p inhibitor, respectively. The cells were then intervened with 30 mmol/L glucose for 24 hours. The expression of BDNF-AS and miR-145-5p were detected by RT-qPCR. Cell proliferation was detected by CCK-8, and apoptosis was detected by flow cytometry. The expression of Bcl-2 and Bax proteins were detected by Western blotting, and the levels of IL-1β and IL-6 in cell culture supernatant were detected by enzyme-linked immunosorbent assay. Dual luciferase reporter gene experiment was used to verify the regulatory relationship of BDNF-AS with miR-145-5p. RESULTS: High glucose promoted the expression of BDNF-AS in HK-2 cells (P<0.05), but inhibited that of miR-145-5p (P<0.05). Interfering with BDNF-AS or overexpression of miR-145-5p decreased the inhibition rate, apoptosis rate and expression of Bax protein, IL-1β and IL-6 of HK-2 cells induced by high glucose (P<0.05), but promoted the expression of Bcl-2 protein (P<0.05). Interfering with miR-145-5p reversed the effect of interfering with BDNF-AS on the proliferation, apoptosis rate and the expression of IL-1β and IL-6 of HK-2 cells induced by high glucose. BDNF-AS could target and down-regulate miR-145-5p. CONCLUSION Interfering with BDNF-AS may promote the proliferation of renal tubular epithelial cells induced by high glucose and inhibit cell apoptosis and the expression of inflammatory factor by down-regulating miR-145-5p.
Apoptosis ; Brain-Derived Neurotrophic Factor/genetics* ; Cell Proliferation ; Epithelial Cells ; Glucose ; Humans ; MicroRNAs/genetics*

A fusion gene called Ig-BDNF, in which brain-derived neurotrophic factor cDNA fused to the 3' end of signal peptide of Ig coding sequence, was constructed by PCR, digested and subcloned into shuttle plasmid pSNAV to obtain a recombinant plasmid pSNAV-Ig-BDNF. Then the plasmid encoding fusion protein was transfected into 293 cell lines and the stably transfected clones were selected with neomycin. AAV1 containing Ig-BDNF fusion gene vectors were obtained by super-infection by Herpes virus. The resultant adeno-associated virus vectors AAV-Ig-BDNF were confirmed by PCR, Western blotting and a sandwich enzyme-linked immunosorbent assay (ELISA) after infection of 293 cell lines. The results indicated that AAV-Ig-BDNF contained the target gene, and infected cells and produced the fusion protein into the supernatant. The content of BDNF in medium per 5x104 cells over a 24 h incubation period reached 1000 pg/mL. With the help of non-replicative adenovirus during AAV-Ig-BDNF infection, the expression of BDNF increased 7-8 fold, and the enhancement of BDNF gene expression was observed in a concentration-dependent manner. These results suggested that a functional AAV-Ig-BDNF was successfully constructed and it offers basis for further study for gene therapy of neural degeneration diseases.
Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Cell Line ; Dependovirus ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Simplexvirus ; genetics ; Transfection

Obesity is a great risk factor for type 2 diabetes and certain types of cancer, which become a major burden for public health worldwide. As a classic complex disease, obesity is regarded as the interaction of genetic and environmental factors. However, it is controversial which of these two factors have greater effect on obesity. Several genetic loci have recently been reported to contribute to the development of obesity reported in genome-wide association study (GWAS) these years. GWAS play an important role in complex disease research and explore the potential effect of genetic variance. To further understand the genetic influence on obesity risk, we reviewed and collected articles on Pubmed for genes that reported in recent GWAS. We summarized the publications in GWAS and found 49 candidate genes, which were strongly suggested to relate to obesity risk in human. Despite the findings of this and other similar, contemporary research projects, much of the single nucleotide polymorphism details and underlying mechanism in this field of study remains, to a great extent, unknown. As a result, future studies are needed for obesity risk in human beings.
Aldose-Ketose Isomerases ; genetics ; Alpha-Ketoglutarate-Dependent Dioxygenase FTO ; Brain-Derived Neurotrophic Factor ; genetics ; Genome-Wide Association Study ; trends ; Humans ; Obesity ; genetics ; physiopathology ; Polymorphism, Single Nucleotide ; Proteins ; genetics

Using the E. coli, we fabricated the gene reconstructed brain derived neurotrophic factor with a fibrin binding domain (FBD-BDNF). We then tested the neurotrophic bioactivity and fibrin-binding ability of the FBD-BDNF. The E. coli was used to express the recombinant protein. The inclusion body was purified with column chromatography and renaturated to construct the right 3D formation. In this study, we successfully fabricated the FBD-BDNF and tested the binding ability and neurotrophic activity. The results demonstrated that FBD-BDNF had special binding ability of fibrin and significant neurotrophic activity for DRG cells. FBD-BDNF could have a promising application prospect in nerve tissue engineering.
Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Fibrin ; metabolism ; Genetic Vectors ; genetics ; Humans ; Protein Binding ; Recombinant Fusion Proteins ; biosynthesis ; genetics

Based on the CX3C chemokine ligand 1(CX3CL1)-CX3C chemokine receptor 1(CX3CR1) axis, this study explored the potential mechanism by which Zuogui Jiangtang Jieyu Formula(ZGJTJY) improved neuroinflammation and enhanced neuroprotective effect in a rat model of diabetes mellitus complicated with depression(DD). The DD rat model was established by feeding a high-fat diet combined with streptozotocin(STZ) intraperitoneal injection for four weeks and chronic unpredictable mild stress(CUMS) combined with isolated cage rearing for five weeks. The rats were divided into a control group, a model group, a positive control group, an inhibitor group, and a ZGJTJY group. The open field test and forced swimming test were used to assess the depression-like behaviors of the rats. Enzyme-linked immunosorbent assay(ELISA) was performed to measure the expression levels of the pro-inflammatory cytokines interleukin-1β(IL-1β) and tumor necrosis factor-α(TNF-α) in plasma. Immunofluorescence staining was used to detect the expression of ionized calcium-binding adapter molecule 1(Iba1), postsynaptic density protein-95(PSD95), and synapsin-1(SYN1) in the hippocampus. Hematoxylin-eosin(HE) staining, Nissl staining, and TdT-mediated dUTP nick end labeling(TUNEL) fluorescence staining were performed to assess hippocampal neuronal damage. Western blot was used to measure the expression levels of CX3CL1, CX3CR1, A2A adenosine receptor(A2AR), glutamate receptor 2A(NR2A), glutamate receptor 2B(NR2B), and brain-derived neurotrophic factor(BDNF) in the hippocampus. Compared with the model group, the ZGJTJY group showed improved depression-like behaviors in DD rats, enhanced neuroprotective effect, increased expression of PSD95, SYN1, and BDNF(P<0.01), and decreased expression of Iba1, IL-1β, and TNF-α(P<0.01), as well as the expression of CX3CL1, CX3CR1, A2AR, NR2A, and NR2B(P<0.01). These results suggest that ZGJTJY may exert its neuroprotective effect by inhibiting the CX3CL1-CX3CR1 axis and activation of hippocampal microglia, thereby improving neuroinflammation and abnormal activation of N-methyl-D-aspartate receptor(NMDAR) subunits, and ultimately enhancing the expression of synaptic-related proteins PSD95, SYN1, and BDNF in the hippocampus.
Rats ; Animals ; Depression/drug therapy* ; Brain-Derived Neurotrophic Factor ; Neuroprotective Agents ; Tumor Necrosis Factor-alpha/metabolism* ; Neuroinflammatory Diseases ; Diabetes Mellitus ; Receptors, Glutamate ; CX3C Chemokine Receptor 1/genetics*

OBJECTIVE: To explore the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus after ovariectomy in mice. METHODS: BDNF levels were detected by immunohistochemistry combined image analysis in hippocampal CA regions and dentate gyrus of ovariectomized mice. RESULTS: The expression of BDNF in hippocampus of mice decreased significantly after the ovariectomy after 4 days. The recovery BDNF expression started 14 days after the ovariectomy and after 28 days, the expression of BDNF in hippocampus recovered to the normal level. CONCLUSION The decrease of estrogen in ovariectomized mice can weaken the expression of BDNF in hippocampus during the early stage.
Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Dentate Gyrus ; metabolism ; Estrogens ; blood ; Female ; Hippocampus ; metabolism ; Mice ; Ovariectomy ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVE: To provide further foundation for the therapy for spine cord injury by transferred mesenchymal stem cells (MSC) expressing the brain-derived neurotrophic factor (BDNF) gene. METHODS: Bone mesenchymal stem cells were isolated from rats and directly cultivated and expanded in vitro. Bone mesenchymal stem cells were divided into 3 groups: a pEGFP-N1- BDNF-transfected group (BDNF group), a pEGFP-N1 transfected group (vector group) and a non-transfected group. Western blot was applied to detect the expression of transgenic bone mesenchymal stem cells. The differentiation of bone mesenchymal stem cells was identified by immunofluorescence. The rates of induction of neuron-like cells among the three cell groups were compared. RESULTS: The cell surface markers of bone mesenchymal stem cells included expression of CD90 (+), CD44 (+), CD34 (-), CD45 (-). The expression of BDNF in the transfected bone mesenchymal stem cells was demonstrated by Western blot. The positive ratio of neuron-like differentiation in the BDNF group was higher than that of both the vector group and the non-transfected group (P<0.05). CONCLUSION BDNF gene plays an important role in promoting bone marrow mesenchymal stem cells to differentiate into neuron-like cells.
Animals ; Bone Marrow Cells ; cytology ; Brain-Derived Neurotrophic Factor ; genetics ; Cell Differentiation ; genetics ; Cells, Cultured ; Female ; Male ; Mesenchymal Stem Cells ; cytology ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVE: To evaluate the role of brain-derived neurotrophic factor (BDNF) in allergic rhinitis of rat. METHOD: Thirty SD rats free of disease were randomly divided into two groups. A model of allergic rhinitis rat was established by using ovalbumin intraperitoneal immunization and intranasal antigen challenge. The nasal mucosa from 18 out of 20 AR models as well as 10 normal controls were studied for expression of BDNF mRNA by reverse transcription-polymerase chain reaction (RT-PCR). RESULT: BDNF/beta-actin ratio in AR models was significantly higher than control (0.49+/-0.07 vs 0.28+/-0.01, P<0.05). CONCLUSION BDNF played an important role in the development of allergic rhinitis of rat.
Animals ; Brain-Derived Neurotrophic Factor ; genetics ; metabolism ; Disease Models, Animal ; Nasal Mucosa ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Rhinitis, Allergic, Perennial ; metabolism

OBJECTIVETo assess the association between brain-derived neurotrophic factor (BDNF) gene Val66Met polymorphism and clinical characteristics of first episode schizophrenia in a Chinese Han population.

METHODSGenotyping of BDNF Val66Met polymorphism was carried out for 135 schizophrenic patients and 483 healthy controls with TaqMan probe technology. The patients' psychotic symptoms were assessed using the positive and negative syndrome scale (PANSS).

RESULTSA significant difference was found in genotype distribution and allelic frequency of the Val66Met polymorphism between the two groups (P< 0.01). In patients, Met homozygotes had a significantly higher score in anxiety/depression factor, cognitive factor and total score of PANSS than Val carriers.

CONCLUSIONBDNF gene Val66Met polymorphism is associated with the pathogenesis of schizophrenia. The Met/Met genotype of BDNF Val66Met variant may be a risk factor for symptoms in first episode schizophrenia patients.

Adolescent ; Adult ; Brain-Derived Neurotrophic Factor ; genetics ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Schizophrenia ; genetics ; Young Adult

AIMTo study the protective effect of adenovirus mediated brain derived neurotrophic factors on nerve injury.

METHODSSH-SY5Y cells were infected by the recombinant Ad-BDNF. In the serum-free condition, the morphological changes were observed, and MTT method was used to examine the trophic effects of different dosages of Ad-BDNF. The whole cells DNA of infected and control groups were extracted to detect the DNA ladder--the marker of apoptosis.

RESULTS AND CONCLUSIONThe adenoviruses mediated BDNF gene promoted the cell survival and differentiation. It also inhibited the serum-deprived induced cell apoptosis.

Adenoviridae ; genetics ; Apoptosis ; Brain-Derived Neurotrophic Factor ; genetics ; Cell Differentiation ; Cell Line, Tumor ; Cell Survival ; Culture Media, Serum-Free ; Gene Transfer Techniques ; Humans ; Neurons