Background: Cytokines are essential cellular modulators of various physiological and pathological activities, including peripheral nerve repair and regeneration. However, the molecular changes of these cellular mediat...Background: Cytokines are essential cellular modulators of various physiological and pathological activities, including peripheral nerve repair and regeneration. However, the molecular changes of these cellular mediators after peripheral nerve injury are still unclear. This study aimed to identify cytokines critical for the regenerative process of injured peripheral nerves.Methods: The sequencing data of the injured nerve stumps and the dorsal root ganglia(DRG) of Sprague-Dawley(SD) rats subjected to sciatic nerve(SN) crush injury were analyzed to determine the expression patterns of genes coding for cytokines. PCR was used to validate the accuracy of the sequencing data.Results: A total of 46, 52, and 54 upstream cytokines were differentially expressed in the SN at 1 day, 4 days, and 7 days after nerve injury. A total of 25, 28, and 34 upstream cytokines were differentially expressed in the DRG at these time points. The expression patterns of some essential upstream cytokines are displayed in a heatmap and were validated by PCR. Bioinformatic analysis of these differentially expressed upstream cytokines after nerve injury demonstrated that inflammatory and immune responses were significantly involved.Conclusions: In summary, these findings provide an overview of the dynamic changes in cytokines in the SN and DRG at different time points after nerve crush injury in rats, elucidate the biological processes of differentially expressed cytokines, especially the important roles in inflammatory and immune responses after peripheral nerve injury, and thus might contribute to the identification of potential treatments for peripheral nerve repair and regeneration.展开更多
Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and foun...Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and found that expression of platelet factor 4 was markedly up-regulated after sciatic nerve injury.Platelet factor is an important molecule in cell apoptosis,diffe rentiation,survival,and proliferation.Further,polymerase chain reaction and immunohistochemical staining confirmed the change in platelet factor 4 in the sciatic nerve at different time points after injury.Enzyme-linked immunosorbent assay confirmed that platelet factor 4 was secreted by Schwann cells.We also found that silencing platelet factor 4 decreased the proliferation and migration of primary cultured Schwann cells,while exogenously applied platelet factor 4 stimulated Schwann cell prolife ration and migration and neuronal axon growth.Furthermore,knocking out platelet factor 4 inhibited the prolife ration of Schwann cells in injured rat sciatic nerve.These findings suggest that Schwann cell-secreted platelet factor 4 may facilitate peripheral nerve repair and regeneration by regulating Schwann cell activation and axon growth.Thus,platelet factor 4 may be a potential therapeutic target for traumatic peripheral nerve injury.展开更多
Cellular senescence and proliferation are essential for wound healing and tissue remodeling.However,senescence-proliferation cell fate after peripheral nerve injury has not been clearly revealed.Here,post-injury gene ...Cellular senescence and proliferation are essential for wound healing and tissue remodeling.However,senescence-proliferation cell fate after peripheral nerve injury has not been clearly revealed.Here,post-injury gene expression patterns in rat sciatic nerve stumps(SRP113121)and L4–5 dorsal root ganglia(SRP200823)obtained from the National Center for Biotechnology Information were analyzed to decipher cellular senescence and proliferation-associated genetic changes.We first constructed a rat sciatic nerve crush model.Then,β-galactosidase activities were determined to indicate the existence of cellular senescence in the injured sciatic nerve.Ki67 and EdU immunostaining was performed to indicate cellular proliferation in the injured sciatic nerve.Both cellular senescence and proliferation were less vigorous in the dorsal root ganglia than in sciatic nerve stumps.These results reveal the dynamic changes of injury-induced cellular senescence and proliferation from both genetic and morphological aspects,and thus extend our understanding of the biological processes following peripheral nerve injury.The study was approved by the Animal Ethics Committee of Nantong University,China(approval No.20190226-001)on February 26,2019.展开更多
Magnesium（Mg） wire has been shown to be biodegradable and have anti-inflammatory properties. It can induce Schwann cells to secrete nerve growth factor and promote the regeneration of nerve axons after central nervo...Magnesium（Mg） wire has been shown to be biodegradable and have anti-inflammatory properties. It can induce Schwann cells to secrete nerve growth factor and promote the regeneration of nerve axons after central nervous system injury. We hypothesized that biodegradable Mg wire may enhance compressed peripheral nerve regeneration. A rat acute sciatic nerve compression model was made, and AZ31 Mg wire（3 mm diameter; 8 mm length） bridged at both ends of the nerve. Our results demonstrate that sciatic functional index, nerve growth factor, p75 neurotrophin receptor, and tyrosine receptor kinase A m RNA expression are increased by Mg wire in Mg model. The numbers of cross section nerve fibers and regenerating axons were also increased. Sciatic nerve function was improved and the myelinated axon number was increased in injured sciatic nerve following Mg treatment. Immunofluorescence histopathology showed that there were increased vigorous axonal regeneration and myelin sheath coverage in injured sciatic nerve after Mg treatment. Our findings confirm that biodegradable Mg wire can promote the regeneration of acute compressed sciatic nerves.展开更多
The repair of peripheral nerve injuries with autologous nerve remains the gold standard （Wang et al., 2005; Yao et al., 2010; Deal et al., 2012; Kriebel et al., 2014; Liu et al., 2014; Tamaki et al., 2014; Yu et al.,...The repair of peripheral nerve injuries with autologous nerve remains the gold standard （Wang et al., 2005; Yao et al., 2010; Deal et al., 2012; Kriebel et al., 2014; Liu et al., 2014; Tamaki et al., 2014; Yu et al., 2014; Zhu and Lou, 2014）. With advances in tissue engineering and biomaterials, tissue-engineered nerve conduits with various biomaterials and structures, such as collagen and chitosan nerve conduits, have already been used in the clinic as alternatives to autologous nerve in the repair of peripheral nerve injury （Wang et al., 2012; Svizenska et al., 2013; Eppenberger et al., 2014; Gu et al., 2014; Koudehi et al., 2014; MoyaDiaz et al., 2014; Novajra et al., 2014; Okamoto et al., 2014; Shea et al., 2014; Singh et al., 2014; Tamaki et al., 2014; Yu et al., 2014）. Therefore, new simple and effective methods展开更多
BACKGROUND： Ceramide galactosyltransferase （CGT） protein and mRNA expression defect can cause the abnormal morphology and slowing conduction velocity of peripheral nerve. Morphologic change and functional disorder ...BACKGROUND： Ceramide galactosyltransferase （CGT） protein and mRNA expression defect can cause the abnormal morphology and slowing conduction velocity of peripheral nerve. Morphologic change and functional disorder of myelin sheath and axon appear when diabetic peripheral neuropathy （DPN） occurs. Whether electroacupuncture at Zusanfi（ST 36） and Shenshu（BL 32） points can enhance the expression of CGT protein and mRNA in the DPN tissue？ OBJECTIVE： To observe the effect of electroacupuncture at Zusanfi and Shenshu points on motor, sensory conduction velocity and CGT mRNA and its protein expression of sciatic nerve in rats with DPN. DESIGN： A randomized and controlled animal experiment SETTING ： Department of Neurology and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine, Shanghai University of Traditional Chinese Medicine. MATERIALS ： Totally 60 healthy male Wistar rats of clean grade, aged 4 month, with body mass of 200 to 220 g, were enrolled in this study. Streptozotocin （STZ, Sigma Company of USA, Batch No. S-0130）. METHODS： This study was carried out in the Animal Experimental Center and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine during February 2005 to March 2006. （1） Fifteen rats were randomly chosen,serving as normal group.AU the other rats were intraperitoneally injected once with STZ to develop experimental diabetic rat models. If fasting blood glucose was ≥ 15 mmol/L,sensory nerve and motor nerve conduction velocity of sciatic nerve was obviously slowed, tail-swaying temperature threshold was increased and myelinated nerve fiber of sciatic nerve changed, DPN models were successful. The successful model rats were randomly assigned into 3 groups： model group, control group（electroacupuncture at non-meridian-non-acupoint）and electroacupuncture group [electroacupuncture at Zusan/i and Shenshu points], with 15 rats each. The rats in the normal group and model group were untouched. In the electroacupuncture group （electroacupuncture at Zusanfi and Shenshu points）, Shenshu point （double） and Zusanfi point （double） were chosen referencing to The Atlas of the Rat＇s Acupoints. G6805- Ⅱ electric acupuncture apparatus was used, and current intensity was controlled at 20 min/time, once every other day, 12 times within 24 days. In the control group, the tip of rat-tail was stimulated, and the concrete procedures were the same as in the electroacupuncture at Zusanfi and Shenshu points. （2） Motor nerve conduction velocity and sensory nerve conduction velocity of rats were detected with neuroelectrophysiology detector in the end of the treatment, and the expressions of CGT protein and mRNA of sciatic nerve were detected with immunohistochemical method and fluorescent quantitative PCR technique. MAIN OUTCOME MEASURES： （1） Motor and sensory nerve conduction velocity. （2） The expression of CGT protein and its mRNA. RESULTS： All the 60 rats entered the stage of result analysis. （1） Comparison of motor and sensory nerve conduction velocity of rats after electroacupuncture： Motor nerve conduction velocity of rats in the model group[（31.37±3.69） m/s], control group [（32.74±5.42） m/s] and electroacupuncture group [（41.30 ±1.15） m/s] was significantly lower than that in the normal group [（41.30±1.15） m/s, P 〈 0.01]; The sensory nerve conduction velocity of rats in the model group[（18.17±9.54） m/s], control group [（21.39±5.61） m/s]and electroacupuncture group [（35.81 ±4.59） m/s] was significantly lower than that in the normal group [（46.38± 6.32） m/s,P 〈 0.01]; The motor and sensory nerve conduction velocity of electroacupuncture group was significantly higher than that in the model group [（38.04±2.01） m/s vs. （32.74±5.42） m/s,（35.81±4.59） m/s vs. （21.39±5.61） m/s,P 〈 0.01]. （2） Comparison of the expression of CGT protein of sciatic nerve of rats： The number of CGT positive cells of sciatic nerve in model group, control group or electroacupuncture group was significantly smaller than that in normal group [（9 770.33±1 461.73）, （10 588.13±1119.52）, （27 518.27± 9 078.29）, （37 769.67±4 021.81）/μm^2,P 〈 0.01]; The number of CGT positive cells of the sciatic nerve in the electroacupuncture group was significantly larger than that in the model group and control group （P 〈 0.01）. The number of CGT positive cells of sciatic nerve was close between control group and electroacupuncture group （P 〉 0.05）. （3） Comparison of CGT mRNA expression of sciatic nerve of rats： Ct value of CGT mRNA of sciatic nerve of rats in the model group,control group and electroacupuncture group was significantly higher than that in the normal group （13.75±2.60,14.81±2.80,11.67±1.75,9.30±0.98, P 〈 0.01 ）; Ct value of CGT mRNA of sciatic nerve of rats in the electroacupuncture group was significantly lower than that in the model group and control group （P 〈 0.01）, and that was close between electroacupuncture group and control group （P 〉 0.05）. CONCLUSION ： Electroacupuncture at Zusanfi and Shenshu points can increase motor and sensory nerve conduction velocity of rats with DPN. It might be associated with up-regulating the expression of CGT mRNA and its protein.展开更多
BACKGROUND: Sodium valproate (VPA) is used to be an effective anti-epileptic drug. VPA possesses the characteristics of penetrating rapidly through the blood-brain barrier (BBB) and increasing levels of Bcl-2 and grow...BACKGROUND: Sodium valproate (VPA) is used to be an effective anti-epileptic drug. VPA possesses the characteristics of penetrating rapidly through the blood-brain barrier (BBB) and increasing levels of Bcl-2 and growth cone-associated protein (GAP) 43 in spinal cord. OBJECTIVE: To observe the effect of VPA on Bcl-2 expression and motor neuronal apoptosis in spinal cord of rats following sciatic nerve transection. DESIGN: Randomized controlled experiment. SETTING: Department of Hand Surgery and Microsurgery, Wuhan Puai Hospital. MATERIALS: A total of 30 male healthy SD rats of clean grade and with the body mass of 180-220 g were provided by Experimental Animal Center of Medical College of Wuhan University. Sodium Valproate Tablets were purchases from Hengrui Pharmaceutical Factory, Jiangsu. METHODS: The experiment was performed in the Central Laboratory of Wuhan Puai Hospital and Medical College of Wuhan University from February to May 2006. Totally 30 rats were randomly divided into two groups: treatment group (n =15) and model group (n =15). Longitudinal incision along backside of right hind limbs of rats was made to expose sciatic nerves, which were sharply transected 1 cm distal to the inferior margin of piriform muscle after nerve liberation under operation microscope to establish sciatic nerve injury rat models. Sodium Valproate Tablets were pulverized and diluted into 50 g/L suspension with saline. On the day of operation, the rats in the treatment group received 6 mL/kg VPA suspension by gastric perfusion, once a day, whereas model group received 10 mL/kg saline by gastric perfusion, once a day. L4-6 spinal cords were obtained at days 1, 4, 7, 14 and 28 after operation, respectively. Terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) technique and immunohistochemical method (SP method) were used to detect absorbance (A) of neurons with positive Bcl-2 expression. Apoptotic rate of cells (number of apoptotic cells/total number of cells×100%) was calculated. MAIN OUTCOME MEASURES: A value of neurons with positive Bcl-2 expression and apoptotic rate in spinal cord of rats in the two groups. RESULTS: A total of 30 SD rats were involved in the result analysis. ①expression of positive Bcl-2 neurons: A value of positive Bcl-2 neurons were 0.71±0.02, 0.86±0.04, 1.02±0.06 at days 4, 7 and 14, respectively after operation in the treatment group, which were obviously higher than those in the model group (0.62±0.03, 0.71±0.05, 0.89±0.04, t = 3.10-4.50, P < 0.05). ②apoptotic result of motor neurons: Apoptotic rate of motor neurons in spinal cord was (6.91±0.89)% and (15.12±2.34)% at days 7 and 14 in the treatment group, which was significantly lower than those in the model group [(9.45±1.61)%, (19.35±0.92)%, t = 2.39, 3.03. P < 0.05]. CONCLUSION: VPA can increase expression of Bcl-2 in spinal cord and reduce neuronal apoptosis in rats following sciatic nerve injury, and has protective effect on motor neuron in spinal cord of rats.展开更多
The regenerative capacity of peripheral nerves is limited after nerve injury.A number of growth factors modulate many cellular behaviors,such as proliferation and migration,and may contribute to nerve repair and regen...The regenerative capacity of peripheral nerves is limited after nerve injury.A number of growth factors modulate many cellular behaviors,such as proliferation and migration,and may contribute to nerve repair and regeneration.Our previous study observed the dynamic changes of genes in L4–6 dorsal root ganglion after rat sciatic nerve crush using transcriptome sequencing.Our current study focused on upstream growth factors and found that a total of 19 upstream growth factors were dysregulated in dorsal root ganglions at 3,9 hours,1,4,or 7 days after nerve crush,compared with the 0 hour control.Thirty-six rat models of sciatic nerve crush injury were prepared as described previously.Then,they were divided into six groups to measure the expression changes of representative genes at 0,3,9 hours,1,4 or 7 days post crush.Our current study measured the expression levels of representative upstream growth factors,including nerve growth factor,brain-derived neurotrophic factor,fibroblast growth factor 2 and amphiregulin genes,and explored critical signaling pathways and biological process through bioinformatic analysis.Our data revealed that many of these dysregulated upstream growth factors,including nerve growth factor,brain-derived neurotrophic factor,fibroblast growth factor 2 and amphiregulin,participated in tissue remodeling and axon growth-related biological processes Therefore,the experiment described the expression pattern of upstream growth factors in the dorsal root ganglia after peripheral nerve injury.Bioinformatic analysis revealed growth factors that may promote repair and regeneration of damaged peripheral nerves.All animal surgery procedures were performed in accordance with Institutional Animal Care Guidelines of Nantong University and ethically approved by the Administration Committee of Experimental Animals,China(approval No.20170302-017)on March 2,2017.展开更多
基金supported by the Postgraduate Research&Practice Innovation Program of Jiangsu Province (KYCX19_2064)the Nantong University Undergraduate Innovation Program (201910304032Z)the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)。
文摘Background: Cytokines are essential cellular modulators of various physiological and pathological activities, including peripheral nerve repair and regeneration. However, the molecular changes of these cellular mediators after peripheral nerve injury are still unclear. This study aimed to identify cytokines critical for the regenerative process of injured peripheral nerves.Methods: The sequencing data of the injured nerve stumps and the dorsal root ganglia(DRG) of Sprague-Dawley(SD) rats subjected to sciatic nerve(SN) crush injury were analyzed to determine the expression patterns of genes coding for cytokines. PCR was used to validate the accuracy of the sequencing data.Results: A total of 46, 52, and 54 upstream cytokines were differentially expressed in the SN at 1 day, 4 days, and 7 days after nerve injury. A total of 25, 28, and 34 upstream cytokines were differentially expressed in the DRG at these time points. The expression patterns of some essential upstream cytokines are displayed in a heatmap and were validated by PCR. Bioinformatic analysis of these differentially expressed upstream cytokines after nerve injury demonstrated that inflammatory and immune responses were significantly involved.Conclusions: In summary, these findings provide an overview of the dynamic changes in cytokines in the SN and DRG at different time points after nerve crush injury in rats, elucidate the biological processes of differentially expressed cytokines, especially the important roles in inflammatory and immune responses after peripheral nerve injury, and thus might contribute to the identification of potential treatments for peripheral nerve repair and regeneration.
基金supported by the National Natural Science Foundation of China,Nos.31730031,32130060the National Natural Science Foundation of China,No.31971276(to JH)+1 种基金the Natural Science Foundation of Jiangsu Province,No.BK20202013(to XG)the Natural Science Foundation of Jiangsu Higher Education Institutions of China(Major Program),No.19KJA320005(to JH)。
文摘Schwann cells in peripheral nerves react to traumatic nerve injury by attempting to grow and regenerate.Howeve r,it is unclear what factors play a role in this process.In this study,we searched a GEO database and found that expression of platelet factor 4 was markedly up-regulated after sciatic nerve injury.Platelet factor is an important molecule in cell apoptosis,diffe rentiation,survival,and proliferation.Further,polymerase chain reaction and immunohistochemical staining confirmed the change in platelet factor 4 in the sciatic nerve at different time points after injury.Enzyme-linked immunosorbent assay confirmed that platelet factor 4 was secreted by Schwann cells.We also found that silencing platelet factor 4 decreased the proliferation and migration of primary cultured Schwann cells,while exogenously applied platelet factor 4 stimulated Schwann cell prolife ration and migration and neuronal axon growth.Furthermore,knocking out platelet factor 4 inhibited the prolife ration of Schwann cells in injured rat sciatic nerve.These findings suggest that Schwann cell-secreted platelet factor 4 may facilitate peripheral nerve repair and regeneration by regulating Schwann cell activation and axon growth.Thus,platelet factor 4 may be a potential therapeutic target for traumatic peripheral nerve injury.
基金supported by the National Natural Science Foundation of China,No.31970968(to SYL)Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)。
文摘Cellular senescence and proliferation are essential for wound healing and tissue remodeling.However,senescence-proliferation cell fate after peripheral nerve injury has not been clearly revealed.Here,post-injury gene expression patterns in rat sciatic nerve stumps(SRP113121)and L4–5 dorsal root ganglia(SRP200823)obtained from the National Center for Biotechnology Information were analyzed to decipher cellular senescence and proliferation-associated genetic changes.We first constructed a rat sciatic nerve crush model.Then,β-galactosidase activities were determined to indicate the existence of cellular senescence in the injured sciatic nerve.Ki67 and EdU immunostaining was performed to indicate cellular proliferation in the injured sciatic nerve.Both cellular senescence and proliferation were less vigorous in the dorsal root ganglia than in sciatic nerve stumps.These results reveal the dynamic changes of injury-induced cellular senescence and proliferation from both genetic and morphological aspects,and thus extend our understanding of the biological processes following peripheral nerve injury.The study was approved by the Animal Ethics Committee of Nantong University,China(approval No.20190226-001)on February 26,2019.
基金supported by the National Natural Science Foundation of China,No.81400528the China Postdoctoral Science Foundation,No.20130390827
文摘Magnesium（Mg） wire has been shown to be biodegradable and have anti-inflammatory properties. It can induce Schwann cells to secrete nerve growth factor and promote the regeneration of nerve axons after central nervous system injury. We hypothesized that biodegradable Mg wire may enhance compressed peripheral nerve regeneration. A rat acute sciatic nerve compression model was made, and AZ31 Mg wire（3 mm diameter; 8 mm length） bridged at both ends of the nerve. Our results demonstrate that sciatic functional index, nerve growth factor, p75 neurotrophin receptor, and tyrosine receptor kinase A m RNA expression are increased by Mg wire in Mg model. The numbers of cross section nerve fibers and regenerating axons were also increased. Sciatic nerve function was improved and the myelinated axon number was increased in injured sciatic nerve following Mg treatment. Immunofluorescence histopathology showed that there were increased vigorous axonal regeneration and myelin sheath coverage in injured sciatic nerve after Mg treatment. Our findings confirm that biodegradable Mg wire can promote the regeneration of acute compressed sciatic nerves.
基金supported by the National High Technology Research and Development Program of China,No.2012AA020502the National Natural Science Foundation of China,No.81171457 and 81371687the Priority of Academic Program Development of Jiangsu Higher Education Institutions
文摘The repair of peripheral nerve injuries with autologous nerve remains the gold standard （Wang et al., 2005; Yao et al., 2010; Deal et al., 2012; Kriebel et al., 2014; Liu et al., 2014; Tamaki et al., 2014; Yu et al., 2014; Zhu and Lou, 2014）. With advances in tissue engineering and biomaterials, tissue-engineered nerve conduits with various biomaterials and structures, such as collagen and chitosan nerve conduits, have already been used in the clinic as alternatives to autologous nerve in the repair of peripheral nerve injury （Wang et al., 2012; Svizenska et al., 2013; Eppenberger et al., 2014; Gu et al., 2014; Koudehi et al., 2014; MoyaDiaz et al., 2014; Novajra et al., 2014; Okamoto et al., 2014; Shea et al., 2014; Singh et al., 2014; Tamaki et al., 2014; Yu et al., 2014）. Therefore, new simple and effective methods
基金the National Nat-ural Science Foundation of Chi-na, No. 30472238 the Grantfrom Bureau of Public Health ofShanghai City, No. 05JC14053Shanghai Key Subjects Con-struction Program, No. T0302
文摘BACKGROUND： Ceramide galactosyltransferase （CGT） protein and mRNA expression defect can cause the abnormal morphology and slowing conduction velocity of peripheral nerve. Morphologic change and functional disorder of myelin sheath and axon appear when diabetic peripheral neuropathy （DPN） occurs. Whether electroacupuncture at Zusanfi（ST 36） and Shenshu（BL 32） points can enhance the expression of CGT protein and mRNA in the DPN tissue？ OBJECTIVE： To observe the effect of electroacupuncture at Zusanfi and Shenshu points on motor, sensory conduction velocity and CGT mRNA and its protein expression of sciatic nerve in rats with DPN. DESIGN： A randomized and controlled animal experiment SETTING ： Department of Neurology and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine, Shanghai University of Traditional Chinese Medicine. MATERIALS ： Totally 60 healthy male Wistar rats of clean grade, aged 4 month, with body mass of 200 to 220 g, were enrolled in this study. Streptozotocin （STZ, Sigma Company of USA, Batch No. S-0130）. METHODS： This study was carried out in the Animal Experimental Center and Central Laboratory, Yueyang Hospital of Traditional Chinese ＆ Western Medicine during February 2005 to March 2006. （1） Fifteen rats were randomly chosen,serving as normal group.AU the other rats were intraperitoneally injected once with STZ to develop experimental diabetic rat models. If fasting blood glucose was ≥ 15 mmol/L,sensory nerve and motor nerve conduction velocity of sciatic nerve was obviously slowed, tail-swaying temperature threshold was increased and myelinated nerve fiber of sciatic nerve changed, DPN models were successful. The successful model rats were randomly assigned into 3 groups： model group, control group（electroacupuncture at non-meridian-non-acupoint）and electroacupuncture group [electroacupuncture at Zusan/i and Shenshu points], with 15 rats each. The rats in the normal group and model group were untouched. In the electroacupuncture group （electroacupuncture at Zusanfi and Shenshu points）, Shenshu point （double） and Zusanfi point （double） were chosen referencing to The Atlas of the Rat＇s Acupoints. G6805- Ⅱ electric acupuncture apparatus was used, and current intensity was controlled at 20 min/time, once every other day, 12 times within 24 days. In the control group, the tip of rat-tail was stimulated, and the concrete procedures were the same as in the electroacupuncture at Zusanfi and Shenshu points. （2） Motor nerve conduction velocity and sensory nerve conduction velocity of rats were detected with neuroelectrophysiology detector in the end of the treatment, and the expressions of CGT protein and mRNA of sciatic nerve were detected with immunohistochemical method and fluorescent quantitative PCR technique. MAIN OUTCOME MEASURES： （1） Motor and sensory nerve conduction velocity. （2） The expression of CGT protein and its mRNA. RESULTS： All the 60 rats entered the stage of result analysis. （1） Comparison of motor and sensory nerve conduction velocity of rats after electroacupuncture： Motor nerve conduction velocity of rats in the model group[（31.37±3.69） m/s], control group [（32.74±5.42） m/s] and electroacupuncture group [（41.30 ±1.15） m/s] was significantly lower than that in the normal group [（41.30±1.15） m/s, P 〈 0.01]; The sensory nerve conduction velocity of rats in the model group[（18.17±9.54） m/s], control group [（21.39±5.61） m/s]and electroacupuncture group [（35.81 ±4.59） m/s] was significantly lower than that in the normal group [（46.38± 6.32） m/s,P 〈 0.01]; The motor and sensory nerve conduction velocity of electroacupuncture group was significantly higher than that in the model group [（38.04±2.01） m/s vs. （32.74±5.42） m/s,（35.81±4.59） m/s vs. （21.39±5.61） m/s,P 〈 0.01]. （2） Comparison of the expression of CGT protein of sciatic nerve of rats： The number of CGT positive cells of sciatic nerve in model group, control group or electroacupuncture group was significantly smaller than that in normal group [（9 770.33±1 461.73）, （10 588.13±1119.52）, （27 518.27± 9 078.29）, （37 769.67±4 021.81）/μm^2,P 〈 0.01]; The number of CGT positive cells of the sciatic nerve in the electroacupuncture group was significantly larger than that in the model group and control group （P 〈 0.01）. The number of CGT positive cells of sciatic nerve was close between control group and electroacupuncture group （P 〉 0.05）. （3） Comparison of CGT mRNA expression of sciatic nerve of rats： Ct value of CGT mRNA of sciatic nerve of rats in the model group,control group and electroacupuncture group was significantly higher than that in the normal group （13.75±2.60,14.81±2.80,11.67±1.75,9.30±0.98, P 〈 0.01 ）; Ct value of CGT mRNA of sciatic nerve of rats in the electroacupuncture group was significantly lower than that in the model group and control group （P 〈 0.01）, and that was close between electroacupuncture group and control group （P 〉 0.05）. CONCLUSION ： Electroacupuncture at Zusanfi and Shenshu points can increase motor and sensory nerve conduction velocity of rats with DPN. It might be associated with up-regulating the expression of CGT mRNA and its protein.
文摘BACKGROUND: Sodium valproate (VPA) is used to be an effective anti-epileptic drug. VPA possesses the characteristics of penetrating rapidly through the blood-brain barrier (BBB) and increasing levels of Bcl-2 and growth cone-associated protein (GAP) 43 in spinal cord. OBJECTIVE: To observe the effect of VPA on Bcl-2 expression and motor neuronal apoptosis in spinal cord of rats following sciatic nerve transection. DESIGN: Randomized controlled experiment. SETTING: Department of Hand Surgery and Microsurgery, Wuhan Puai Hospital. MATERIALS: A total of 30 male healthy SD rats of clean grade and with the body mass of 180-220 g were provided by Experimental Animal Center of Medical College of Wuhan University. Sodium Valproate Tablets were purchases from Hengrui Pharmaceutical Factory, Jiangsu. METHODS: The experiment was performed in the Central Laboratory of Wuhan Puai Hospital and Medical College of Wuhan University from February to May 2006. Totally 30 rats were randomly divided into two groups: treatment group (n =15) and model group (n =15). Longitudinal incision along backside of right hind limbs of rats was made to expose sciatic nerves, which were sharply transected 1 cm distal to the inferior margin of piriform muscle after nerve liberation under operation microscope to establish sciatic nerve injury rat models. Sodium Valproate Tablets were pulverized and diluted into 50 g/L suspension with saline. On the day of operation, the rats in the treatment group received 6 mL/kg VPA suspension by gastric perfusion, once a day, whereas model group received 10 mL/kg saline by gastric perfusion, once a day. L4-6 spinal cords were obtained at days 1, 4, 7, 14 and 28 after operation, respectively. Terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP-biotin nick end labeling (TUNEL) technique and immunohistochemical method (SP method) were used to detect absorbance (A) of neurons with positive Bcl-2 expression. Apoptotic rate of cells (number of apoptotic cells/total number of cells×100%) was calculated. MAIN OUTCOME MEASURES: A value of neurons with positive Bcl-2 expression and apoptotic rate in spinal cord of rats in the two groups. RESULTS: A total of 30 SD rats were involved in the result analysis. ①expression of positive Bcl-2 neurons: A value of positive Bcl-2 neurons were 0.71±0.02, 0.86±0.04, 1.02±0.06 at days 4, 7 and 14, respectively after operation in the treatment group, which were obviously higher than those in the model group (0.62±0.03, 0.71±0.05, 0.89±0.04, t = 3.10-4.50, P < 0.05). ②apoptotic result of motor neurons: Apoptotic rate of motor neurons in spinal cord was (6.91±0.89)% and (15.12±2.34)% at days 7 and 14 in the treatment group, which was significantly lower than those in the model group [(9.45±1.61)%, (19.35±0.92)%, t = 2.39, 3.03. P < 0.05]. CONCLUSION: VPA can increase expression of Bcl-2 in spinal cord and reduce neuronal apoptosis in rats following sciatic nerve injury, and has protective effect on motor neuron in spinal cord of rats.
基金supported by the Natural Science Foundation of Jiangsu Higher Education Institutions of China(Major Program),No.16KJA310005(to SYL)the Natural Science Foundation of Nantong City of China,No.JC2018058(to TMQ)the Priority Academic Program Development of Jiangsu Higher Education Institutions of China
文摘The regenerative capacity of peripheral nerves is limited after nerve injury.A number of growth factors modulate many cellular behaviors,such as proliferation and migration,and may contribute to nerve repair and regeneration.Our previous study observed the dynamic changes of genes in L4–6 dorsal root ganglion after rat sciatic nerve crush using transcriptome sequencing.Our current study focused on upstream growth factors and found that a total of 19 upstream growth factors were dysregulated in dorsal root ganglions at 3,9 hours,1,4,or 7 days after nerve crush,compared with the 0 hour control.Thirty-six rat models of sciatic nerve crush injury were prepared as described previously.Then,they were divided into six groups to measure the expression changes of representative genes at 0,3,9 hours,1,4 or 7 days post crush.Our current study measured the expression levels of representative upstream growth factors,including nerve growth factor,brain-derived neurotrophic factor,fibroblast growth factor 2 and amphiregulin genes,and explored critical signaling pathways and biological process through bioinformatic analysis.Our data revealed that many of these dysregulated upstream growth factors,including nerve growth factor,brain-derived neurotrophic factor,fibroblast growth factor 2 and amphiregulin,participated in tissue remodeling and axon growth-related biological processes Therefore,the experiment described the expression pattern of upstream growth factors in the dorsal root ganglia after peripheral nerve injury.Bioinformatic analysis revealed growth factors that may promote repair and regeneration of damaged peripheral nerves.All animal surgery procedures were performed in accordance with Institutional Animal Care Guidelines of Nantong University and ethically approved by the Administration Committee of Experimental Animals,China(approval No.20170302-017)on March 2,2017.
