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OBJECTIVE@#To explore the mechanism of electroacupuncture (EA) in promoting recovery of the facial function with the involvement of autophagy, glial cell line-derived neurotrophic factor (GDNF), and phosphatidylinositol-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway.@*METHODS@#Seventy-two male Sprague-Dawley rats were randomly allocated into the control, sham-operated, facial nerve injury (FNI), EA, EA+3-methyladenine (3-MA), and EA+GDNF antagonist groups using a random number table, with 12 rats in each group. An FNI rat model was established with facial nerve crushing method. EA intervention was conducted at Dicang (ST 4), Jiache (ST 6), Yifeng (SJ 17), and Hegu (LI 4) acupoints for 2 weeks. The Simone's 10-Point Scale was utilized to monitor the recovery of facial function. The histopathological evaluation of facial nerves was performed using hematoxylin-eosin (HE) staining. The levels of Beclin-1, light chain 3 (LC3), and P62 were detected by immunohistochemistry (IHC), immunofluorescence, and reverse transcription-polymerase chain reaction, respectively. Additionally, IHC was also used to detect the levels of GDNF, Rai, PI3K, and mTOR.@*RESULTS@#The facial functional scores were significantly increased in the EA group than the FNI group (P<0.05 or P<0.01). HE staining showed nerve axons and myelin sheaths, which were destroyed immediately after the injury, were recovered with EA treatment. The expressions of Beclin-1 and LC3 were significantly elevated and the expression of P62 was markedly reduced in FNI rats (P<0.01); however, EA treatment reversed these abnormal changes (P<0.01). Meanwhile, EA stimulation significantly increased the levels of GDNF, Rai, PI3K, and mTOR (P<0.01). After exogenous administration with autophagy inhibitor 3-MA or GDNF antagonist, the repair effect of EA on facial function was attenuated (P<0.05 or P<0.01).@*CONCLUSIONS@#EA could promote the recovery of facial function and repair the facial nerve damages in a rat model of FNI. EA may exert this neuroreparative effect through mediating the release of GDNF, activating the PI3K/mTOR signaling pathway, and further regulating the autophagy of facial nerves.
Subject(s)
Rats , Male , Animals , Rats, Sprague-Dawley , Electroacupuncture , Phosphatidylinositol 3-Kinase/metabolism , Facial Nerve Injuries/therapy , Phosphatidylinositol 3-Kinases/metabolism , Beclin-1 , Glial Cell Line-Derived Neurotrophic Factor , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Autophagy , Mammals/metabolismABSTRACT
ObjectiveTo investigate the expression of glial cell line-derived neurotrophic factor （GDNF） and androgen receptor （AR） in testicular peritubular cells （TPCs） of cryptorchidism mouse models and explore the theoretical significance of cryptorchidism-induced spermatogenesis dysfunction. MethodsA total of 30 five-week-old male ICR rats were divided randomly by using random number table method into 6 groups. Cryptorchidism was surgically induced in 3 randomly selected groups and the other 3 groups underwent sham surgery as the control groups. On days 4， 7 and 14 after surgery， we harvested the mice testes of the 3 groups and their corresponding control groups， then measured the testicular volumes， analyzed the testicular histopathology and detected the mRNA and protein expression levels of AR and GDNF in TPCs by immunofluorescence， real-time PCR and Western blot. ResultsIn normal control groups， on days 4， 7 and 14 after surgery， the testicular volumes were （125.58±19.22） mm3，（123.45±20.12） mm3， （140.09±13.62） mm3 ， respectively. Clear layers of spermatogenic cells were well arranged and abundant sperm cells were found. Peritubular cells were morphologically homogeneous， with slim-spindle appearance and normal cell thickness. The mRNA expression levels of AR were 1.00±0.05， 1.06±0.07 and 1.19±0.13； GDNF mRNA 1.00±0.04， 1.09±0.05， and 1.10±0.07. The protein expression levels of AR were 1.01±0.01， 0.79±0.02 and 1.01±0.04； GDNF protein （18.68±0.43） pg/mL， （14.39±0.36） pg/mL and （16.88±0.37） pg/mL. In cryptorchidism groups， on days 4， 7 and 14 after surgery， the testicular volumes were （115.64±3.91） mm3， （69.51±14.97） mm3 and （44.86±5.56） mm3， respectively. Spermatogenic cells were disorganized， seminiferous tubules were disrupted， peritubular cells shrank， bent and fractured. The mRNA expression levels of AR were 0.76±0.06， 0.53±0.04， and 0.29±0.02； GDNF mRNA 0.72±0.05， 0.42±0.02 and 0.30±0.03. The protein expression levels of AR were 0.54±0.02， 0.98±0.04 and 0.31±0.01； GDNF protein （8.50±0.34） pg/mL， （17.44±0.32） pg/mL and （6.83±0.34） pg/mL. Statistically significant differences （P < 0.05） were found in 7-day and 14-day testicular volumes between control and cryptorchidism groups but not in the 4-day testicular volume （P > 0.05）. Testicular volumes， AR and GDNF mRNA and protein expression in control groups had no statistically significant difference （P > 0.05）， while those in cryptorchidism groups showed a trend of gradual decline in the amount and the differences between groups were statistically significant （P < 0.05）. ConclusionsIn surgery-induced cryptorchidism mice， after the induction， the expression of AR and GDNF in TPCs showed a gradual decrease over time. AR and GDNF play a major role in mediating the TPCs damage in cryptorchidism. This study provides a theoretical basis for mechanism researches of cryptorchidism-induced spermatogenesis dysfunction.
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Glial cell line-derived neurotrophic factor (GDNF) is an important functional protein derived from enteric glial cells. It plays an important role in the enteric nervous system, such as nutritional neurons, promoting synaptic remodeling and anti-inflammation. The role of GDNF in the progression of gastrointestinal diseases has received more attention gradually. This article reviewed GDNF and its ligands, related signaling pathway, correlation with intestinal homeostasis and clinical application.
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ObjectiveTo investigate whether Xiayuxue decoction exerts an anti-liver fibrosis effect by inhibiting glial cell line-derived neurotrophic factor (GDNF). MethodsA total of 24 C57BL/6 mice were randomly divided into control group, model group, and Xiayuxue decoction group. The mice in the model group and the Xiayuxue decoction group were given intraperitoneal injection of 10% CCl4, and those in the Xiayuxue decoction group were given 0.4678 g/kg Xiayuxue decoction by gavage since week 4. The liver function parameters alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured, and liver histopathology was observed. Immunohistochemistry was used to measure the protein expression of alpha-smooth muscle actin (α-SMA) and GDNF. GFP-Col-HSC and human primary hepatic stellate cells (HSCs) were treated with GDNF (10 ng/ml), and HSC activation was measured. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the control group, the model group had significant increases in the levels of ALT and AST, and compared with the model group, the Xiayuxue decoction group had significant reductions in the levels of ALT and AST (all P＜0.01). Liver histopathology showed that the model group had marked inflammatory cell infiltration and formation of fibrous septa by proliferated collagen fibers, and the Xiayuxue decoction group had loose fibrous septa and alleviated inflammatory cell infiltration. Immunohistochemistry showed that compared with the control group, the model group had significant increases in the expression of α-SMA and GDNF (both P＜0.01), which were observed in fibrous septa, and compared with the model group, the Xiayuxue decoction group had significant reductions in the expression of α-SMA and GDNF (both P＜0.05). Western blotting showed that the control group had relatively low expression of GDNF in liver tissue, the formation of liver fibrosis at week 6 of CCl4 modeling, and an around 10-fold increase in the expression of GDNF, and the Xiayuxue decoction group had significantly inhibited protein expression of GDNF (P＜0.01); there were significant increases in the expression of α-SMA and collagen type I α1 (Col1) in mice with liver fibrosis, with significant reductions in α-SMA and Col1 after treatment with Xiayuxue decoction (all P＜0.01). The in vitro experiment showed that GDNF induced the significant increases in the protein expression of α-SMA and Col1 in HSCs, which was significantly inhibited by Xiayuxue decoction (all P＜0.01). ConclusionThe expression of GDNF is significantly upregulated in the formation of liver fibrosis. GDNF can induce HSC activation, and Xiayuxue decoction can exert an anti-liver fibrosis effect by inhibiting GDNF.
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BACKGROUND: Glial cell line derived neurotrophic factor (GDNF) plays an important role in inducing differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro and promoting neurological function recovery in rats with spinal cord injury. OBJECTIVE: To observe potential molecular mechanisms of differentiation of BMSCs overexpressing GDNF gene and promoting neurological function recovery after spinal cord injury in rats. METHODS: (1) BMSCs transfected with recombinant target gene adenovirus were divided into Ad-GDNF-GFP transfection group, Ad-GFP transfection group and non-transfection group. Microtubule-associated protein 2 and neuron-specific enolase expression levels were detected by immunofluorescence in each group. Western blot assay was used to detect the expression of GDNF, Wnt3a and Wnt7a protein in each group. (2) The rat spinal cord injury model was prepared by modified Allen method. The 45 Sprague-Dawley rat models were randomly divided into three groups. GDNF-BMSCs, BMSCs and PBS were transplanted into the site of spinal cord injury. The motor function recovery of rats was evaluated 4 weeks after operation. The morphological changes of spinal cord were observed by hematoxylin-eosin staining. The local neuron-specific enolase, Synapsin I and glial fibrillary acidic protein were analyzed with immunohistochemistry. The expression levels of Bcl-2 and tumor necrosis factor-α protein were detected by western blot assay. RESULTS AND CONCLUSION: (1) BMSCs overexpressing GDNF gene could differentiate into neuron-like cells and express neuron-specific enolase and microtubule-associated protein 2 in vitro in the Ad-GDNF-GFP transfection group. The expression of Wnt3a and Wnt7a protein was significantly higher in the Ad-GDNF-GFP transfection group than in the Ad-GFP transfection group and non-transfection group. (2) The Basso, Beattie and Bresnahan score in GDNF-BMSCs group was significantly increased and the stenosis area was significantly reduced at 4 weeks after transplantation. The expression of glial fibrillary acidic protein and tumor necrosis factor-α in GDNF-BMSCs group was significantly lower than that in BMSCs and PBS transplantation groups, but the expression levels of neuron-specific enolase, Synapsin I and Bcl-2 were significantly higher than those in the BMSCs and PBS transplantation groups. (3) Wnt signaling pathway participates in the procession of differentiating into mature neurons derived from BMSCs overexpressing GDNF gene. After transplantation, the effects of BMSCs transplantation on spinal cord injury were improved by decreasing local inflammatory reaction, apoptosis and glial scar formation and promoting axonal regeneration.
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While hallmarks of rodent spermatogonia stem cell biomarkers' heterogeneity have recently been identified, their stage and subset distributions remain unclear. Furthermore, it is currently difficult to accurately identify subset-specific SSC marker distributions due to the poor nuclear morphological characteristics associated with fixation in 4% paraformaldehyde. In the present study, testicular cross-sections and whole-mount samples were Bouin fixed to optimize nuclear resolution and visualized by immunohistochemistry (IHC) and immunofluorescence (IF). The results identified an expression pattern of PLZFhighc-KITpos in A1 spermatogonia, while A2-A4-differentiating spermatogonia were PLZFlowc-KITpos. Additionally, this procedure was used to examine asymmetrically expressing GFRA1 and PLZF clones, asymmetric Apr and false clones were distinguished based on the presence or absence of TEX14, a molecular maker of intercellular bridges, despite having identical nuclear morphology and intercellular distances that were <25 μm. In conclusion, this optimized Bouin fixation procedure facilitates the accurate identification of spermatogonium subsets based on their molecular profiles and is capable of distinguishing asymmetric and false clones. Therefore, the findings presented herein will facilitate further morphological and functional analysis studies and provide further insight into spermatogonium subtypes.
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Spinal cord injury (SCI) causes axonal damage and demyelination, neural cell death, and comprehensive tissue loss, resulting in devastating neurological dysfunction. Neural stem/progenitor cell (NSPCs) transplantation provides therapeutic benefits for neural repair in SCI, and glial cell line-derived neurotrophic factor (GDNF) has been uncovered to have capability of stimulating axonal regeneration and remyelination after SCI. In this study, to evaluate whether GDNF would augment therapeutic effects of NSPCs for SCI, GDNF-encoding or mock adenoviral vector-transduced human NSPCs (GDNF-or Mock-hNSPCs) were transplanted into the injured thoracic spinal cords of rats at 7 days after SCI. Grafted GDNF-hNSPCs showed robust engraftment, long-term survival, an extensive distribution, and increased differentiation into neurons and oligodendroglial cells. Compared with Mock-hNSPC- and vehicle-injected groups, transplantation of GDNF-hNSPCs significantly reduced lesion volume and glial scar formation, promoted neurite outgrowth, axonal regeneration and myelination, increased Schwann cell migration that contributed to the myelin repair, and improved locomotor recovery. In addition, tract tracing demonstrated that transplantation of GDNF-hNSPCs reduced significantly axonal dieback of the dorsal corticospinal tract (dCST), and increased the levels of dCST collaterals, propriospinal neurons (PSNs), and contacts between dCST collaterals and PSNs in the cervical enlargement over that of the controls. Finally grafted GDNF-hNSPCs substantially reversed the increased expression of voltage-gated sodium channels and neuropeptide Y, and elevated expression of GABA in the injured spinal cord, which are involved in the attenuation of neuropathic pain after SCI. These findings suggest that implantation of GDNF-hNSPCs enhances therapeutic efficiency of hNSPCs-based cell therapy for SCI.
Subject(s)
Animals , Humans , Rats , Axons , Cell Death , Cell Movement , Cell- and Tissue-Based Therapy , Cicatrix , Demyelinating Diseases , gamma-Aminobutyric Acid , Glial Cell Line-Derived Neurotrophic Factor , Hyperalgesia , Myelin Sheath , Neuralgia , Neurites , Neuroglia , Neurons , Neuropeptide Y , Paraplegia , Pyramidal Tracts , Regeneration , Spinal Cord Injuries , Spinal Cord , Therapeutic Uses , Transplants , Voltage-Gated Sodium ChannelsABSTRACT
Objective@#To determine the effect of a motor-specific neurotrophic factor, glial-derived neurotrophic factor (GDNF) on motor nerve regeneration.@*Methods@#We used a nerve conduit filled with a fibrin-based delivery system that provided controlled release of GDNF during nerve regeneration. The motor branch of the rat femoral nerve was used to assess motor nerve regeneration across a 5-mm gap. Four experimental groups (n=5) were evaluated. These included GDNF with the fibrin-based delivery system (GDNF-DS group), fibrin alone(fibrin group), empty conduit (negative control group), and nerve isograft (positive control group). Nerves were harvested at 5 weeks for analysis by histomorphometry and electron microscopy.@*Results@#At 5 mm distal to the conduit or isografts, the GDNF-DS group was not significantly different from the nerve isograft group in the following histomorphometric measures: total nerve fibers, percentage of neural tissue, and nerve density. The number of nerve fibers (respectively: 1 744±274 , 1 481±288) and the percentage of nerve tissue [(14.2±3.9)%, (11.0±2.2)%] in theisograft group and the GDNF-DS group were significantly higher than that in the fibrin group and the empty conduit group [(respectively: 538±93, 535±96) and the percentage of nerve tissue respectively: (4.3±1.6)%, (3.7±0.9)%]. There were no differences in fiber width among all groups. By electron microscopy, the GDNF-DS and isograft groups also demonstrated more organized nerve architecture than the fibrin alone and empty conduit groups.@*Conclusions@#The delivery of GDNF from the fibrin-based delivery system promotes motor nerve regeneration at a level similar to an isograft in the femoral motor nerve model. This study gives insight into the potential beneficial role of GDNF in the treatment of motor nerve injuries.
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Objective To determine the effect of a motor-specific neurotrophic factor,glial-derived neurotrophic factor (GDNF) on motor nerve regeneration.Methods We used a nerve conduit filled with a fibrin-based delivery system that provided controlled release of GDNF during nerve regeneration.The motor branch of the rat femoral nerve was used to assess motor nerve regeneration across a 5-mm gap.Four experimental groups (n =5) were evaluated.These included GDNF with the fibrin-based delivery system (GDNFDS group),fibrin alone(fibrin group),empty conduit (negative control group),and nerve isograft (positive control group).Nerves were harvested at 5 weeks for analysis by histomorphometry and electron microscopy.Results At 5 mm distal to the conduit or isografts,the GDNF-DS group was not significantly different from the nerve isograft group in the following histomorphometric measures:total nerve fibers,percentage of neural tissue,and nerve density.The number of nerve fibers (respectively:1 744 ± 274,1 481 ± 288)and the percentage of nerve tissue [(14.2 ± 3.9) ％,(11.0 ± 2.2) ％] in theisograft group and the GDNFDS group were significantly higher than that in the fibrin group and the empty conduit group [(respectively:538 ± 93,535 ± 96) and the percentage of nerve tissue respectively:(4.3 ± 1.6) ％,(3.7 ± 0.9) ％].There were no differences in fiber width among all groups.By electron microscopy,the GDNF-DS and isograft groups also demonstrated more organized nerve architecture than the fibrin alone and empty conduit groups.Conclusions The delivery of GDNF from the fibrin-based delivery system promotes motor nerve regeneration at a level similar to an isograft in the femoral motor nerve model.This study gives insight into the potential beneficial role of GDNF in the treatment of motor nerve injuries.
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While hallmarks of rodent spermatogonia stem cell biomarkers' heterogeneity have recently been identified, their stage and subset distributions remain unclear. Furthermore, it is currently difficult to accurately identify subset-specific SSC marker distributions due to the poor nuclear morphological characteristics associated with fixation in 4% paraformaldehyde. In the present study, testicular cross-sections and whole-mount samples were Bouin fixed to optimize nuclear resolution and visualized by immunohistochemistry (IHC) and immunofluorescence (IF). The results identified an expression pattern of PLZFhighc-KITpos in A1 spermatogonia, while A2-A4-differentiating spermatogonia were PLZFlowc-KITpos. Additionally, this procedure was used to examine asymmetrically expressing GFRA1 and PLZF clones, asymmetric Apr and false clones were distinguished based on the presence or absence of TEX14, a molecular maker of intercellular bridges, despite having identical nuclear morphology and intercellular distances that were <25 μm. In conclusion, this optimized Bouin fixation procedure facilitates the accurate identification of spermatogonium subsets based on their molecular profiles and is capable of distinguishing asymmetric and false clones. Therefore, the findings presented herein will facilitate further morphological and functional analysis studies and provide further insight into spermatogonium subtypes.
Subject(s)
Animals , Male , Mice , Cell Differentiation , Fluorescent Antibody Technique , Gene Expression Regulation/genetics , Glial Cell Line-Derived Neurotrophic Factor Receptors/genetics , Immunohistochemistry , Mice, Inbred C57BL , Promyelocytic Leukemia Zinc Finger Protein/genetics , Proto-Oncogene Proteins c-kit/genetics , Seminiferous Tubules/cytology , Spermatogenesis , Spermatogonia/metabolism , Testis/cytology , Tissue Fixation , Transcription Factors/geneticsABSTRACT
Glial cell line-derived neurotrophic factor (GDNF) has been reported to be involved in negatively regulating the effects of addictive disorders. The objective of this study was to investigate alterations in the levels of GDNF in patients with Internet gaming disorder (IGD) and to assess the relationship between GDNF levels and the severity of IGD indices. Nineteen male patients with IGD and 19 sexmatched control subjects were evaluated for alteration of plasma GDNF levels and for relationship between GDNF levels and clinical characteristics of Internet gaming, including the Young's Internet Addiction Test (Y-IAT). The GDNF levels were found to be significantly low in patients with IGD (103.2±62.0 pg/mL) compared with the levels of controls (245.2±101.6 pg/mL, p<0.001). GDNF levels were negatively correlated with Y-IAT scores (Spearman's rho=-0.645, p=<0.001) and this negative correlation remained even after controlling for multiple variables (r=-0.370, p=0.048). These findings support the assumed role of GDNF in the regulation of IGD.
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Humans , Male , Case-Control Studies , Glial Cell Line-Derived Neurotrophic Factor , Immunoglobulin D , Internet , Neuroglia , Pilot Projects , PlasmaABSTRACT
Objective To observe the changes of protein and mRNA expression levels of brain derived neurotrophic factor(BDNF) and glial cell line derived neurotrophic factor (GDNF) in the prefrontal cortex,hippocampus and striatum of the attention deficit hyperactivity disorder(ADHD) model animal,and to explore the relationship between BDNF,GDNF and ADHD.Methods Junior spontsneously hyoertensive rat (SHR) rats were used as ADHD model group and Wistar Kyoto rats(WKY) as control group.The protein and mRNA expression levels of BDNF and GDNF in prefrontal cortex,hippocampus and striatum were detected by Western blot and RT-PCR methods.Results The expression of BDNF and GDNF protein was (0.561±0.095) and (0.507±0.155) in the prefrontal cortex,(0.606±0.054) and (0.457±0.198) in hippocampus and (0.558±0.191) and (0.554±0.267) in striatum of SHR rats.The expression of BDNF and GDNF protein of WKY rats were (0.855±0.211) and (0.891±0.244) in the prefrontal cortex,(0.827±0.163) and (0.898±0.207) in hippocampus and (0.998±0.220) and (0.955±0.146) in striatum.The expression of BDNF and GDNF mRNA was (0.636±0.141) and (0.544±0.161) in the prefrontal cortex,(0.996±0.196) and (0.563±0.302) in hippocampus and (0.664±0.145) and (0.689±0.049) in striatum of SHR rats.The expression of BDNF and GDNF mRNA of WKY rats were (0.861±0.232) and (0.840 ±0.124) in prefrontal cortex,(1.372±0.423) and (1.180±0.441) in hippocampus,and (1.107±0.185)and (0.950±0.247) in striatum.The protein and mRNA expression levels of BDNF and GDNF in prefrontal cortex,hippocampus and striatum of ADHD rats were significantly lower than those in the control group (all P＜0.05).Conclusion The low expression of BDNF and GDNF in the prefrontal cortex,hippocampus and striatum of SHR rats may be related to the pathophysiology of ADHD.
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Objective: To discuss the protective effect of velvet antler polypeptides (VAP) combined with Schwann cells (SCs) modified by glial cell line derived neurotrophic factor (GDNF) gene on the apoptosis of spinal cord neurons induced by beta-amyloid 25-35 (Aβ25-35). Methods: The spinal cord cells of fetal mice were prepared and the spinal cord neurons in logarithmic growth phase were taken; the apoptosis of spinal cord neurons was induced by Aβ25-35. The spinal cord neurons were divided into normal cell group (the normal spinal cord neurons), induced apoptosis group (the apoptotic spinal cord neurons induced by Aβ25-35, SCs group (the apoptotic spinal cord neurons induced by Aβ25-35 + SCs), GDNF group (the apoptotic spinal cord neurons induced by Aβ25-35 + GDNF), SCs + GDNF group (the apoptotic spinal cord neurons induced by Aβ25-35 + GDNF-transfected SCs) and VAP combination group (the apoptotic spinal cord neurons induced by Aβ25-35 + VAP combined with GDNF-transfected SCs). Flow cytometry was used to detect the apoptotic rates of spinal cord neurons in various groups; immunohistochemical staining was used to detect the number of caspase-3 positive cells in the spinal cord neurons in various groups. Results; After suspension inoculation of fetal spinal cord neurons, most of them were round at the initial stage. The flow cytometry results showed that compared with induced apoptosis group, the apoptotic rates of spinal cord neurons in SCs, GDNF, SCs + GDNF, and VAP combination groups were decreased (P<0.05). Compared with SCs + GDNF group, the apoptotic rate of spinal cord neurons in VAP combination group was decreased (P<0. 05). The immunohistochemistry results showed that the expression of caspase-3 in spinal cord neurons in various groups could be found. There were no significant differences in the number of caspase-3 positive cells and cell staining between SCs group, GDNF group and SCs + GDNF group; but the number of caspase-3 positive cells in SCs group, GDNF group and SCs + GDNF group were significantly higher than that in induced apoptosis group (P<0. 05). Conclusion: VAP combined with GDNF-transfected SCs has the protective effect on the apoptosis of spinal cord cells by reducing the expression of caspase-3 in spinal cord neurons.
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Objective To evaluate the changes in the expression of artemin in skin around the inci-sion during remifentanil-induced hyperalgesia in the rats with incisional pain. Methods Thirty-two healthy male Sprague-Dawley rats, aged 10-12 weeks, weighing 250-280 g, were divided into 4 groups (n = 8 each) using a random number table: control group (group C), incisional pain group (group I), remifen-tanil group (group R) and incisional pain plus remifentanil group (group I+R). Remifentanil was intrave-nously infused for 60 min at a rate of 1 μg·kg-1 ·min-1 in group R. In group I, the model of incisional pain was established, and the equal volume of normal saline was infused for 60 min via the tail vein at the same time. In group I+R, the model of incisional pain was established, and remifentanil was infused for 60 min via the tail vein at a rate of 1 μg·kg-1 ·min-1 at the same time. The equal volume of normal saline was infused for 60 min via the tail vein in group C. Mechanical paw withdrawal threshold (MWT) and ther-mal paw withdrawal latency (TWL) were measured at 24 h before infusion of remifentanil or normal saline and 2, 6, 24 and 48 h after the end of infusion (T0-4 ). Rats were sacrificed following the last measurement of pain threshold, and ipsilateral plantar skin was removed for detection of the expression of artemin protein and mRNA (by fluorescent quantitative real-time polymerase chain reaction or Western blot). Results Compared with group C, MWT was significantly decreased and TWL was shorten at T1-4 , and the expression of artemin protein and mRNA in plantar skin was up-regulated in R, I and I+R groups (P<0. 01). Compared with R and I groups, MWT was significantly decreased and TWL was shorten at T1-4 , and the ex-pression of artemin protein and mRNA in plantar skin was up-regulated in group I+R (P<0. 01). Conclu-sion The peripheral mechanism by which remifentanil induces hyperalgesia may be related to up-regulated expression of artemin in skin around the incision in the rats with incisional pain.
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OBJECTIVE: Some clinical studies have found alterations in the levels of serum brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) after applying antidepressant treatment in patients with major depressive disorder (MDD). We evaluated the serum BDNF and GDNF levels before and after 12 weeks of antidepressant treatment in MDD outpatients. METHODS: Serum BDNF and GDNF levels were measured in 23 female MDD outpatients at baseline and after 12 weeks of treatment. The severity of depression was measured with the Hamilton Depression Rating Scale-17 (HAMD-17). Remission of MDD to the treatment was defined as a posttreatment HAMD-17 score of <7. RESULTS: Among MDD patients, 19 (82.6%) subjects were in mild to moderate depression. The whole MDD patients had significantly higher serum BDNF and GDNF levels at baseline than those after 12 weeks of antidepressant treatment. The baseline serum BDNF and GDNF levels did not significantly between the remission and nonremission groups. The significant alteration in both BDNF and GDNF levels after antidepressant treatment were observed in patients with remission. CONCLUSION: The present study suggests that the baseline serum BDNF and GDNF levels are higher than the posttreatment levels in some mild-to-moderate MDD outpatients and the significant alteration in BDNF and GDNF level after treatment were observed in patients with remission.
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Female , Humans , Brain-Derived Neurotrophic Factor , Depression , Depressive Disorder, Major , Glial Cell Line-Derived Neurotrophic Factor , OutpatientsABSTRACT
Aim To explore the effects of prenatal caf-feine exposure (PCE) on fetal renal growth retardation and corticosterone on the gene expression of metanephric mesenchyme stem cells.Methods Pregnant Wistar rats were administered with caffeine (30,120 mg ·kg-1) from gestational day 9 to 20.Female fetal kidney samples were collected for morphological observation and gene expression examination.The metanephric mesenchyme stem cells were harvested for cell culture,and renal related genes were detected after the treatment of corticosterone with different concentrations (250,500,1 000 μg · L-1) for 24 hours.Results Compared with the control group,the fetal kidneys in the PCE group displayed an enlarged Bowman's space and a shrunken glomerular tuft,accompanied with the repression of the gene expression of glial-cell-line-derived neurotrophic factor/tyrosine kinase receptor (GDNF/c-Ret) signaling pathway.The GDNF/c-Ret signaling pathway and angiotensin Ⅱ receptor type 1 (AT1R)/AT2R expression of metanephric mesenchyme stem cells also decreased in corticosterone groups.Conclusions PCE may induce dysplasia of female fetal kidneys.The potential mechanism is related to the repression of the gene expression of AT1R/AT2R and GDNF/c-Ret signaling pathway by PCE mediated by corticosterone in utero.
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Objective To study the relationship between executive function and glial cell line-de-rived neurotrophic factor(GDNF) in patients with obsessive-compulsive disorder(OCD). Methods Totally 64 patients with OCD and 61 healthy controls were enrolled. The levels of serum GDNF were measured by en-zyme linked immunosorbent assay (ELISA). Wisconsin card sorting test (WCST) was used to assess the ex-ecutive function of the subjects. Yale-Brown obsessive compulsive scale ( Y-BOCS) was used to assess ob-sessive-compulsive symptoms. Results The patients with OCD(62. 67±8. 48)showed significantly poorer performance than healthy controls on the correct score of WCST(71. 16±7. 24)(P<0. 05),but the errors and non-persistent errors scores(52. 81±8. 39,31. 05±8. 46)were significantly higher than that in healthy con-trols (44. 79±7. 69,26. 57±7. 76)(P<0. 05). The level of serum GDNF in OCD group ((5. 64±1. 01) pg/ml)was significantly lower than that in control group ((6. 99±0. 94) pg/ml). There was a negative cor-relation between the number of non-persistent errors and the level of GDNF in OCD group( r=-0. 304,P=0. 015). The correct number and classification of WCST were negatively correlated with the scores of Y-BOCS(t=-0. 546,-0. 758,P<0. 05),the error of WCST were positively correlated to the scores of Y-BOCS(t=0. 616,P<0. 05). Conclusion These findings suggest that patients with OCD have executive dysfunction. The level of GDNF may be involved in the pathogenesis of OCD,which may be associated with the executive dysfunction in OCD patients.
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Objective To investigate gender differences of plasma glial cell line-derived neurotro-phic factor (GDNF) levels in patients with major depressive disorder (MDD). Methods MDD subjects (male 20,female 36) and healthy controls (HCs) (male 35,female 45) were divided into four groups by gender. Plasma levels of GDNF were measured and compared in different gender groups. The clinical symp-tom severity of MDD patients was evaluated by 17-item Hamilton Depression Scale (HAMD-17) and Hamil-ton Anxiety Scale (HAMA-17). Results (1)The plasma GDNF level in male patients with major depres-sive disorder (( 1. 55 ± 0. 43 ) pg/ml ) was significantly lower than that in healthy controls (( 1. 86 ± 0. 50)pg/ml,F=4. 64,P=0. 036). There was no significant difference in GDNF level between female de-pression patients((1.62±0.46)pg/ml)) and female healthy control((1. 64±0. 48)pg/ml,F=0. 18,P=0. 672). In HCs,the GDNF level of male was significantly higher than that of female((1. 86±0. 50)pg/ml, (1. 64±0. 48)pg/ml,F=2. 04,P=0. 045). There was no significant difference in GDNF level between male and female patients(P>0. 05). (2) GDNF level in male patients with major depressive disorder was nega-tively correlated with HAMA score(r=-0. 388,P=0. 034). Conclusion The expression of GDNF is affect-ed by sex factors,which may be related to the different pathogenesis of MDD.
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Objective:To investigate the effect of pilose antler polypeptide combined with Schwann cells modified by glial cell line-derived neurotrophic factor (GDNF)gene on the proliferation of human bone marrow mesenchymal stem cells (BMSCs)in vitro .Methods:According to the conventional method,the bone marrow (10 mL)was extracted from the healthy volunteers and was inoculated into the culture flask.The primary cultured cells were completely fused.The BMSCs were harvested at the 3rd generation and the cells were adjusted to 5 × 106 mL-1 . 4 μL GDNF gene modified Schwann cells was added into GDNF group,4 μL (10 mg· L-1 )PAP combined with GDNF gene modified Schwann cells was added into combination group,and only same amount of medium was added into control group.The proliferative activities,cell nuclear antigen (PCNA)levels and apoptotic rates of BMSCs in various groups were detected by enzyme-linked immunosorbent assay,ELISA method and Annexin Ⅴ-FIFC/PI cell apoptosis detection kit,respectively.Results:After primary culture for 48 h,most of the cells adhered to the wall,and the morphology of the cells changed into polygonal shape and few of them showed spindle.The passaged cells showed spindle spindle,the cells were confirmed as BMSCs,and all of them were non-hematopoietic stem cells.Compared with control group,the proliferative activities and the PCNA level of the BMSCs in GDNF group and combination group were increased (P <0.05)and the apoptotic rates were decreased (P <0.05);compared with GDNF group,the proliferative activity and the PCNA level of the BMSCs in combination group were increased (P < 0.05 )and the apoptotic rate was decreased (P < 0.05 ).Conclusion:PAP combined with Schwann cells modified by GDNF gene can promote the proliferation of human BMSCs in vitro .
ABSTRACT
Objective To evaluate the role of glial cell line-derived neurotrophic factor family re-ceptor alpha3(GFRα3)in the expression and membrane trafficking of transient receptor potential melasta-tin 8(TRPM8)in the dorsal root ganglion(DRG)during cold hyperalgesia in rats with neuropathic pain (NP). Methods Thirty-two healthy adult male Sprague-Dawley rats, aged 10-12 weeks, weighing 250-280 g, in which intrathecal catheters were successfully implanted, were divided into 4 groups(n=8 each) using a random number table: sham operation plus GFRα3 dsRNA group(Sham+dsRNA group), sham operation plus GFRα3 siRNA group(group Sham+siRNA), NP plus GFRα3 dsRNA group(group NP+dsRNA)and NP plus GFRα3 siRNA group(group NP+siRNA). NP was produced by chronic constriction injury to the sciatic nerve. At 10-30 days after operation, GFRα3 dsRNA 10 μg∕20 μl was intrathecally injected once a day for 4 consecutive days in Sham+dsRNA and NP+dsRNA groups, and 10 μg∕20 μl GFRα3 siRNA, of which the sense strand was modified with 2′-O-methyl and 5′-cholesterol, was intrathe-cally injected once a day for 4 consecutive days in Sham+siRNA and NP+siRNA groups. The number of paw lifts on the cold plate, mechanical paw withdrawal threshold(MWT)and thermal paw withdrawal latency (TWL)were measured on 1 day before operation and 10, 11, 12, 13(before intrathecal injection)and 14 days after operation. The rats were sacrificed after the last behavioral testing, and ipsilateral DRGs of the lumbar segment(L4-6)were dissected for detection of the expression of GFRα3 and TRPM8 in total and membrane proteins by Western blot, and the ratio of TRPM8 expression in the membrane protein to that in the total protein(m∕t ratio)was calculated. Results Compared with group Sham+dsRNA, the number of paw lifts on the cold plate was significantly increased, the MWT was decreased, and TWL was shortened after operation in NP+dsRNA and NP+siRNA groups, the expression of GFRα3 and TRPM8 in total and membrane proteins was significantly up-regulated, and m∕t ratio was increased in group NP+dsRNA, and the expression of GFRα3 in DRGs was significantly down-regulated(P<001), and no significant change was found in the expression of TRPM8 in total and membrane proteins or m∕t ratio in group NP+siRNA(P>005). Compared with group NP+dsRNA, the number of paw lifts on the cold plate was significantly de-creased, the expression of GFRα3 and TRPM8 in total and membrane proteins was down-regulated, m∕t ra-tio was decreased(P<001), and no significant change was found in MWT or TWL in group NP+siRNA (P>005). Conclusion GFRα3 in DRGs can up-regulate the expression of TRPM8 and enhance the membrane trafficking of TRPM8, which may be involved in the maintenance mechanism of cold hyperalgesia in rats with NP.