Neuroprotective role of Tinospora cordifolia extract in streptozotocin induced neuropathic pain

Abstract Diabetic Neuropathy (DN) is one of the prevailing micro vascular complications of diabetes which can be characterized by neuropathic pain. Streptozotocin (STZ) induced diabetes in the rat has been increasingly used as a model of painful diabetic neuropathy. STZ injection leads to neurotoxicity of peripheral nerves that leads to development of Peripheral Diabetic Neuropathy in rat model. The present study was aimed at exploring the protective role of Tinospora cordifolia extract in STZ induced neurotoxicity and evaluating mechanisms responsible for attenuating neuropathic pain. Neuropathic pain markers like hyperalgesia, allodynia and motor deficits were assessed before STZ injection and after the treatment with 250 mg/kg and 500 mg/kg dose of Tinospora cordifolia. Oxidative stress markers, NGF expression in sciatic nerve were observed after seven weeks treatment. Our results demonstrated that seven weeks treatment with Tinospora cordifolia leaf extract significantly relieved thermal hyperalgesia and allodynia by increasing the antioxidant enzyme levels, decreasing the lipid peroxidation and by increasing the Nerve growth factor (NGF) expression in diabetic rat sciatic nerves. Our findings highlighted the beneficial effects of oral administration of Tinospora cordifolia extract in attenuating diabetic neuropathic pain, possibly through a strong antioxidant activity and by inducing NGF m RNA in sciatic nerves.

Effect of electroacupuncture at "Zusanli" (ST 36) on duodenal mast cells, NGF and NTRK1 in rats with functional dyspepsia / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) on duodenal mast cells, nerve growth factor (NGF) and neurotrophic tyrosine kinase receptor type 1 (NTRK1), and to explore the mechanism of electroacupuncture at Zusanli (ST 36) on functional dyspepsia (FD).@*METHODS@#Sixty SPF-grade 10-day-old SD rats were randomly divided into a normal group, a model group, a ketotifen group and an EA group, 15 rats in each group. The FD model was prepared by iodoacetamide combined with rat tail clamping method in the model group, the ketotifen group and the EA group. The rats in the ketotifen group were injected intraperitoneally with ketotifen (1 mg•kg-1•d-1) for 7 days; the rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), with disperse-dense wave, frequency of 2 Hz/50 Hz and intensity of 0.5 mA, 20 min each time, once a day for 14 days. The gastric emptying rate and small intestinal propulsion rate in each group were observed; the morphology of duodenal mucosa was observed by HE staining; the toluidine blue staining was used to observe the number and degranulation of mast cells in duodenal mucosa; the protein and mRNA expressions of NGF, NTRK1 in duodenum were detected by Western blot and real-time PCR; the level of interleukin-1β (IL-1β) in duodenum was measured by ELISA.@*RESULTS@#Compared with the normal group, the gastric emptying rate and small intestinal propulsion rate in the model group were decreased (P<0.01); compared with the model group, the gastric emptying rate and small intestinal propulsion rate in the ketotifen group and the EA group were increased (P<0.01); the small intestinal propulsion rate in the EA group was higher than that in the ketotifen group (P<0.01). In the model group, local defects in duodenal mucosa were observed with a small amount of inflammatory cell infiltration; no obvious abnormality was found in duodenal mucosa of the other groups. Compared with the normal group, the mast cells of duodenal mucosa in the model group were increased significantly with significant degranulation; compared with the model group, the mast cells of duodenal mucosa in the ketotifen group and the EA group were decreased significantly, and the degranulation was not obvious. Compared with the normal group, the protein and mRNA expressions of NGF, NTRK1 as well as the level of IL-1β in duodenum in the model group were increased (P<0.01); compared with the model group, the protein and mRNA expressions of NGF, NTRK1 as well as the levels of IL-1β in duodenum in the ketotifen group and the EA group were decreased (P<0.01, P<0.05); compared with the ketotifen group, the mRNA expression of NGF, as well as the protein and mRNA expressions of NTRK1 in duodenum in the EA group were decreased (P<0.05, P<0.01).@*CONCLUSION@#EA at "Zusanli" (ST 36) could inhibit the activation of duodenal mast cells and regulate the expressions of NGF and its receptor to improve the low-grade inflammatory response of duodenum, resulting in treatment effect on FD.

Gastrodin injection alleviates lung injury caused by focal cerebral ischemia in rats through NGF/TrkA pathway-mediated activation of the anti-inflammatory pathway / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic mechanism of gastrodin injection for alleviating lung injury caused by focal cerebral ischemia in rats and the role of the NGF-TrkA pathway in mediating this effect.@*METHODS@#Forty SD rats were equally randomized into normal group, sham-operated group, model group and gastrodin group, and in the latter two groups, rat models of focal cerebral ischemia were established by embolization of the right middle cerebral artery. After successful modeling, the rats were treated with intraperitoneal injection of gastrodin injection at the daily dose of 10 mg/kg for 14 days. After the treatment, the wet/dry weight ratio of the lung tissue was determined, the pathological changes in the lung tissue were observed using HE staining, and the levels of IL-10 and TNF-α in the arterial blood were detected with ELISA. The expressions of NF-κB p65 and TNF-α in the lung tissue were detected with Western blotting, and the expressions of NGF and TrkA were detected using immunohistochemical staining and Western blotting.@*RESULTS@#Compared with the normal control and sham-operated groups, the rats in the model group showed obvious inflammatory lung injury, significantly increased wet/ dry weight ratio of the lungs (P < 0.01), increased TNF-α level in arterial blood (P < 0.01), and significantly up-regulated protein expressions of NF-κB p65 (P < 0.01), TNF-α (P < 0.01), NGF (P < 0.05) and TrkA(P < 0.05) in the lung tissue. Treatment with gastrodin injection obviously alleviated lung inflammation, decreased the wet/dry weight ratio of the lungs (P < 0.05), and significantly lowered TNF-α level (P < 0.01) and increased IL-10 level in the arterial blood in the rat models (P < 0.01); gastrodin injection also significantly decreased the protein expressions of NF-κB p65 and TNF-α (P < 0.05) and up-regulated the expressions of NGF and TrkA in the lung tissue of the rats (P < 0.05).@*CONCLUSION@#The NGF/TrkA pathway may participate in cerebral ischemia-induced inflammatory lung injury, which can be obviously alleviated by gastrodin through the activation of the anti-inflammatory pathway mediated by the NGF/TrkA pathway.

The neuroprotective effect of rhubarb on MCAO model rats / 中国药理学通报

Aim To study the neuroprotective effect of rhubarb extract on MCAO model rats and explore its mechanism of action. Methods Forty-five SPF male Sprague-Dawley rats were randomly divided into sham group, MCAO group, and MCAO + rhubarb group. MCAO model was prepared by silk plug method, and rhubarb extract was administered at a concentration of 200 mg · kg

Effect of Tongma Qining Prescription on Sciatic Nerve Injury in Rats / 中国药学杂志

OBJECTIVE: To study the effect and mechanism of Tongma Qining prescription(TQP) on improving sciatic nerve injury (SNI) in rats. METHODS: The SNI model rats were replicated by using hemostatic forceps. The model rats were randomly divided into the model group, the mecobalamine group (mecobalamine 6×10-4 g•kg-1), the high-dose group (30 g •kg-1), the medium-dose group (15 g•kg-1) and the low-dose group (7.5 g•kg-1), and the sham operation group was set(10 mice per group). After continuous intragastric administration for 6 weeks, sciatic nerve index (SFI), sensory conduction velocity (SNCV) and hemodynamic indexes were detected in each group. HE staining were employed to detect the pathological alterations of sciatic nerve tissues. Then the expression of BDNF, NGF and VEGF in sciatic nerve were detected by IHC-P, the protein expression of NGF and VEGF were detected by Western blot. RESULTS: Tongma Qining prescription was able to alleviate sciatic nerve tissue injury in SNI rats, significantly improved SFI and SNCV (P<0.01), observably reduced the hematocrit, plasma viscosity and value of whole blood viscosity (high shearing and low sheating)(P<0.05), and markedly upregulated the expression level of BDNF, NGF, VEGF and the protein expression level of NGF, VEGF (P<0.01). CONCLUSION: The Tongma Qining prescription can significantly improve SNI in rats. Its action mechanism may be related to improving blood rheology, promoting angiogenesis and protecting nerve cells.

Effect of electroacupuncture on expressions of Gas7 and NGF in arcular nucleus of rats with focal cerebral ischemia / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) on the expressions of growth arrest-specific protein 7 (Gas7) and nerve growth factor (NGF) in arcuate nucleus (ARC) of rats with focal cerebral ischemia and explore the potential action mechanism of EA in treatment of focal cerebral ischemia.@*METHODS@#A total of 50 SD rats were randomized into 4 groups, named a normal group ( =12), a sham-operation group ( =12), a model group ( =14) and an EA group ( =12). In the model group and the EA group, the thread embolization method was adopted to duplicate the model of the right middle cerebral arterial embolism. In the sham-operation group, the skin of the neck was opened and sutured without any other intervention. In the EA group, EA was applied to "Baihui" (CV 20) and "Zusanli" (ST 36) on the left side, once a day, 30 min each time, consecutively for 21 days, while there was no any intervention in the normal group, the sham-operation group and the model group. Using the immunohistochemistry (IHC) method and Western blot method, the expressions of Gas7 and NFG of ARC on the ischemic side were determined. Using Nissle staining, the morphological changes in ARC neurons were observed.@*RESULTS@#The results of Nissle staining showed that there was no significant change in the morphology of ARC neurons in the normal group and the sham-operation group. In the model group, the volume of neuron cells was atrophied obviously and the cells were arranged irregularly. In the EA group, the morphology of ARC neuron was similar to the normal group. The results of IHC and Western blot indicated that the expressions of immunoreactive neurons and protein of Gas7 and NGF in ARC of the rats in the model group were increased obviously as compared with the normal group and the sham-operation group and the expressions in the EA group were further enhanced as compared with the model group (all <0.05).@*CONCLUSION@#Gas7 and NGF may be participated in the compensatory process of partial protection of the body in the patients with focal cerebral ischemia. EA up-regulates the expressions of Gas7 and NGF in ARC, which may be one of the neuroprotective mechanisms of EA in treatment of cerebral ischemia.

Hypericum perforatum chronic treatment affects cognitive parameters and brain neurotrophic factor levels

Objective: To evaluate the effects of Hypericum perforatum (hypericum) on cognitive behavior and neurotrophic factor levels in the brain of male and female rats. Methods: Male and female Wistar rats were treated with hypericum or water during 28 days by gavage. The animals were then subjected to the open-field test, novel object recognition and step-down inhibitory avoidance test. Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and glial cell-line derived neurotrophic factor (GDNF) levels were evaluated in the hippocampus and frontal cortex. Results: Hypericum impaired the acquisition of short- and long-term aversive memory in male rats, evaluated in the inhibitory avoidance test. Female rats had no immediate memory acquisition and decreased short-term memory acquisition in the inhibitory avoidance test. Hypericum also decreased the recognition index of male rats in the object recognition test. Female rats did not recognize the new object in either the short-term or the long-term memory tasks. Hypericum decreased BDNF in the hippocampus of male and female rats. Hypericum also decreased NGF in the hippocampus of female rats. Conclusions: The long-term administration of hypericum appears to cause significant cognitive impairment in rats, possibly through a reduction in the levels of neurotrophic factors. This effect was more expressive in females than in males.

Depressive symptoms and neurotrophin levels in ostomy patients / Sintomas depressivos e níveis de neurotrofina em pacientes com ostomia

ABSTRACT Objective: The aim of the present study was to investigate the depressive symptoms and changes in neurotrophins (BDNF, NGF, NT-3), and cortisol levels in serum of peripheral blood from ostomy patients compared to healthy control group. Methods: We evaluated ostomy (n = 29) and healthy control (n = 30) patients. The neurotrophin (BDNF, NGF, NT-3), and cortisol levels were assessed by ELISA in serum of peripheral blood. Depressive symptoms were defined based on the Hamilton Depression Rating Scale (HDRS), and major depression disorder was based on clinical interviews and was confirmed with the Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I). Results: The results showed a significant decrease in BDNF levels and, a significant increase in NT-3 levels in serum of peripheral blood from ostomy patients when compared to healthy controls. The levels of NGF and cortisol showed no significant differences between groups. The depressive symptom evaluations by HDRS demonstrated a significant increase in ostomy patients when compared to healthy controls. The major depression disorder diagnosis by SCID-I showed no significant difference between groups. Conclusion: Our results suggest ostomy triggers significant depressive symptoms and alterations in neurotrophins levels in serum of peripheral blood samples collected from these patients.

RESUMO Objetivo: O objetivo do presente estudo foi investigar os sintomas depressivos e alterações nos níveis de neurotrofinas (BDNF, NGF, NT-3) e cortisol em soro de sangue periférico de pacientes ostomizados em comparação com grupo controle saudável. Métodos: Foram avaliados pacientes ostomizados (n = 29) e controles saudáveis (n = 30). Os níveis de neurotrofinas (BDNF, NGF, NT-3) e cortisol foram avaliados por kit ELISA em soro de sangue periférico. Os sintomas depressivos foram definidos com base na Hamilton Depression Rating Scale (HDRS), e o transtorno depressivo maior foi baseado em entrevistas clínicas e confirmado pela Structured Clinical Interview for DSM-IV Axis I Disorders (SCID-I). Resultados: Os resultados mostraram diminuição significativa nos níveis de BDNF e aumento significativo nos níveis de NT-3 no soro de sangue periférico de pacientes ostomizados quando comparados com controles saudáveis. Os níveis de NGF e cortisol não apresentaram diferenças significativas entre os grupos. As avaliações dos sintomas depressivos pela HDRS demonstraram aumento significativo em pacientes ostomizados quando comparados com controles saudáveis. O diagnóstico de transtorno depressivo maior pela SCID-I não mostrou diferença significativa entre os grupos. Conclusão: Nossos resultados sugerem que a ostomia desencadeia sintomas depressivos significativos e alterações nos níveis de neurotrofinas no soro de sangue periférico coletadas desses pacientes

Application of Brachial Plexus Block Guided by B Ultrasound in Operation of Ulna and Radius Fracture in Children / 医学研究杂志

Objective To investigate the effect of B-guided brachial plexus block anesthesia on postoperative cognitive function and serum neurotrophic factor (NGF) and transforming growth factor-β (TGF-β) levels in children with ulnar and radius fractures.Methods From February 2014 to January 2017,we selected 78 children with ulnar radius fractures in our hospital randomly and divided them into observation group and control group,each group 39 cases.The control group was given brachial plexus block anesthesia under the position of nerve stimulation,and the observation group was subjected to B-guided brachial plexus block anesthesia.Anesthesia-related indicators,anesthetic effects and complications were observed and analyzed between the two groups,and hemodynamics parameters of brachial plexus block after 0.5h,cognitive function and serum NGF and TGF-β levels before and after operation were compared.Results Compared with the control group,anesthesia onset,recovery room stay time in the observation group were shorter and the duration of analgesia was longer (P ＜ 0.05).The excellent and good rate of anesthesia in the observation group 97.44％ (38/39) was higher than that in the control group (76.92％,30/39).The incidence of complications in the observation group 5.12％ (2/39) was much lower than that of the control group (23.08％,9/39,P ＜ 0.05).There was no significant difference in systolic peak velocity between the two groups (P ＞ 0.05).Compared with the control group,the end of the diastolic velocity and blood flow of brachial plexus block after 0.5h were higher,resistance and pulsatility index were lower (P ＜ 0.05).There was no significant difference in preoperative cognitive function,serum NGF and TGF-β levels between the two groups before operation (P ＞ 0.05).After operation,compared with the control group,the cognitive function,serum NGF and TGF-β levels were higher in the observation group (P ＜ 0.05).Conclusion Anesthesia effect of B-guided brachial plexus block anesthesia is significant in children with ulnar radius fractures.The incidence of complications is low,which can significantly improve the ulnar arterial hemodynamics,significantly increased serum NGF,TGF-β levels,contribute to postoperative fracture healing,and will not affect the cognitive function of children after surgery.

Effects of aripiprazole on clinical symptoms and serum neurotrophic factor levels in patients with schizophrenia / 中国神经精神疾病杂志

Objective To explore the effects of aripiprazole on clinical symptoms and neurotrophic factor levels in patients with schizophrenia. Methods Forty patients with schizophrenia and 40 normal controls were included in the study. The clinical symptoms of patients receiving aripiprazole only for 12 weeks were evaluated by using the Positive and Negative Syndrome Scale (PANSS). Stroop Color-Word Test (SCWT), Continuous Performance Test, Digit-Symbol Coding Test and Trail Making Test-A were used to evaluate the cognitive function both in patients and controls. Serum levels of Nerve Growth Factor (NGF), Brain Derived Neurotrophic Factor (BDNF) and Neurotrophin 3 (NT-3) were measured using enzyme linked immunosorbent assay. Results The clinical scores, cognitive function and levels of neurotrophic factors were different before and after treatment (P<0.01). And those were significantly lower in patients than in control group (P<0.05). Before treatment, BDNF was negatively correlated with PANSS negative symptom score (r=-0.362, P=0.022);NGF was related to the total score of PANSS (r=0.332, P=0.037) and positive symptoms (r=0.401, P=0.010); NT-3 was associated with negative symptom scores (r=-0.376, P=0.017) and SCWT-color words (r=0.332, P=0.037) in patient group. After treatment, the increase in BDNF was correlated with the reduction in PANSS total score (r=0.371, P=0.018), negative symptom score (r=0.345, P=0.029) and general pathology score (r=0.342, P=0.031). There was a correlation of the increase of NGF with the decrease of PANSS total scores (r=0.437, P=0.005) and with positive symptom scores (r=0.357, P=0.024). Conclusion Treatment with Aripiprazole can improve the clinical symptoms and cognitive functiona impairments in patients with schizophrenia, which may be related to the increase in serum levels of BDNF, NGF and NT-3.

Whole-tissue 3D imaging reveals intra-adipose sympathetic plasticity regulated by NGF-TrkA signal in cold-induced beiging

Sympathetic arborizations act as the essential efferent signals in regulating the metabolism of peripheral organs including white adipose tissues (WAT). However, whether these local neural structures would be of plastic nature, and how such plasticity might participate in specific metabolic events of WAT, remains largely uncharacterized. In this study, we exploit the new volume fluorescence-imaging technique to observe the significant, and also reversible, plasticity of intra-adipose sympathetic arborizations in mouse inguinal WAT in response to cold challenge. We demonstrate that this sympathetic plasticity depends on the cold-elicited signal of nerve growth factor (NGF) and TrkA receptor. Blockage of NGF or TrkA signaling suppresses intra-adipose sympathetic plasticity, and moreover, the cold-induced beiging process of WAT. Furthermore, we show that NGF expression in WAT depends on the catecholamine signal in cold challenge. We therefore reveal the key physiological relevance, together with the regulatory mechanism, of intra-adipose sympathetic plasticity in the WAT metabolism.

Study of acupuncture combined with rat nerve growth factor on neurobehavioral ability of cerebral palsy infant rats and its brain tissue growth and metabolism associated proteins / 中国针灸

<p><b>OBJECTIVE</b>To study the effects of acupuncture combined with rat nerve growth factor (NGF) on the cerebral palsy infant rats and the proteins which associated with growth, apoptosis and metabolism.</p><p><b>METHODS</b>Seventy infant rats were selected, Fifty infant rats of which were made the cerebral palsy infant model by the ligation of unilateral carotid artery for cerebral ischemia and oxygen-deficient environment, then the 30 model rats were randomly divided into a model group, a NGF group and a combined group, 10 rats in each group. Twenty infant rats were used in the sham-operated group and the blank control group, 10 rats in each group. The treatment was not given in the blank control group. The rats in the sham-operated group were cut the neck skin and separated the left carotid artery, and then sutured and disinfected the wound. The intraperitoneal injection of NGF (2000 U•kg•d) was used in the NGF group. Based on the injection in the NGF group, acupuncture was used in the combined group, once a day, and the acupoints were "Baihui" (GV 20), left "nieⅠ" (extra), "Dazhui" (GV 14), "Jizhong" (extra), "Quchi" (LI 11), "Yongquan" (KI 1), "Hegu" (LI 4), "Zhoujie" (extra) and "Xiqianxue" (extra). The same volume of saline was intraperitoneally injected in the model group for continuous 14 days. Neurobehavioral ability score was evaluated after treatment. TUNEL were conducted to detect the brain cell apoptosis rate and the expressions of apoptosis associated gene Bax, Bcl-2 and Casp3 were detected by PCR. The level of nerve growth associated protein (GAP-43) and energy metabolism-related protein monocarboxylate transporter protien 1(MCT 1) were detected by Western blot.</p><p><b>RESULTS</b>After intervention, the neurobehavioral ability of baby rats in the blank control and sham-operated group was normal, but there was various degrees of abnormity in the model group, NGF group and combined group. The scores of neurobehavioral ability of the combined group and NGF group were better than those of the model control (all <0.05), and the scores in the combined group was better than those in the NGF group (all <0.05). The left brain cell apoptosis rate, expressions of Bax and Casp3 in the combined group and NGF group were lower and the expressions of Bcl-2 were higher than those of the model group (all <0.05), with more obvious results of Bax and Gasp3 in the combined group than those in the NGF group (all <0.05). The protein levels of GAP-43 and MCT 1 in the combined group and NGF group were higher than those in the model group (all <0.05), with higher expressions in the combined group compared with those in the NGF group (both <0.05).</p><p><b>CONCLUSION</b>Acupuncture combined with NGF could improve the neurobehavioral ability of cerebral palsy infant rats, inhibit the nerve cell apoptosis and improve the brain tissue injure and energy metabolism by up-regulating the expressions of GAP-43 and MCT 1.</p>

Effects of electroacupuncture at "Changqiang" (GV 1) on expression of nerve growth factor and brain derived neurotrophic factor in rats after acute spinal cord injury / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) at"Changqiang"(GV 1) on expression of nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) in rats after acute spinal cord injury (ASCI), and to explore the mechanism of EA at"Changqiang"(GV 1) on ASCI.</p><p><b>METHODS</b>Twenty-four adult female SD rats were randomly divided into an EA group, a model group and a sham operation group, 8 rats in each one. The rats in the sham operation group were treated with laminectomy to expose the spinal cord without any strike. The rats in the model group and EA group were treated with modified Allen's method to establish ASCI model. After model was established, the rats in the EA group were treated with EA at"Changqiang"(GV 1), once a day for continuous 7 days. The rats in the sham operation group and model group were treated with immobilization, once a day, without any other interventions. The basso beattie bresnahan (BBB) was evaluated 1, 3, 5, 7 days after operation. 7 days after operation, the rats were sacrificed with perfusion and the spinal cord was embedded with paraffin. The morphological changes of spinal cord and neuron were observed by Nissl's staining method; the expressions of NGF and BDNF were detected by immune fluorescence method.</p><p><b>RESULTS</b>3 days, 5 days and 7 days after operation, the BBB scores in the EA group were higher than those in the model group (<0.05, <0.01). The Nissl's staining indicated the gray matter of spinal cord was butterfly-shaped with complete structure and clear boundaries between the gray and white matter; the tabby-shaped Nissl bodies were observed in cytoplasm. There were incomplete gray nucleus, big and saturate local stasis plaque. Compared with the model group, the smallerarea of blood stasis, less severity of neuron edema, better morphology of neuron and no vacuole change were observed in the EA group. The immune fluorescence results indicated the expressions of NGF and BDNF in the model group and EA group were higher than those in the sham operation group (all <0.01); the expressions of NGF and BDNF in the EA group were higher than those in the model group (both <0.01).</p><p><b>CONCLUSION</b>EA at"Changqiang"(GV 1) could improve the expression of NGF and BDNF and increase the score of BBB in rats with ASCI, which is beneficial to the repair of ASCI.</p>

Neuroprotective effects and mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats / 药物评价研究

Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.

Neuroprotective effects and mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats / 药物评价研究

Objective To investigate the neuroprotective mechanisms of Cerebroprotein Hydrolysate for Injection (Ⅰ) on vascular dementia in rats.Method The rat vascular dementia model was prepared using an improved two-vessel occlusion method,and the common carotid artery was only isolated but not blocked in sham group.Rats were randomly divided into sham group,model group,Cerebroprotein Hydrolysate for Injection (Ⅰ) groups with low,medium and high dose (5,10,20 mg/kg) and Cerebroprotein Hydrolysate Injection group (Cerebrolysin,Positive drug,10 mg/kg).The drug was administered by iv injection of rat tail vein once a day for two weeks,while the same volume of saline was administered in sham and model group.At the end of administration,the plasma was collected through abdominal aorta to separate serum,and rat cortex was isolated to prepare homogenate.The levels of nerve growth factor (NGF) and insulin-like growth factor 2 (IGF-2) in serum and level of gamma-aminobutyric acid (GABA) in cortex were detected by ELISA.Level of glutamate (Glu) in cortex of VaD rats was detected by colorimetry.Results Compared with model group,levels of NGF and IGF-2 in the serum of VaD rats and level of GABA in cortex were significantly increased,while level of Glu in cortex was significantly decreased after administration of Cerebroprotein Hydrolysate for Injection (Ⅰ).The increased IGF-2 and GABA levels by Cerebroprotein Hydrolysate for Injection (Ⅰ) were significantly higher than that of Cerebrolysin at same dose.Conclusion The mechanisms underlying the increased leaming and memory ability of VaD rats by Cerebroprotein Hydrolysate for Injection (Ⅰ),are possibly related to the increased levels of NGF and IGF-2 in body and a regulation of the balance between excitatory and inhibitory amino acid neurotransmitters.

The effects of BMP9 combined with NGF on the osteogenic differentiation of C3H10T1/2 mesenchymal stem cells / 实用口腔医学杂志

Objective:To investigate the effects of BMP9 combined with NGF on the osteogenic differentiation of mesenchymal stem cells(MSCs).Methods:Recombinant BMP9 adenovirus was transfected into C3H10T1/2 cells.The cells were treated by GFP,NGF,BMP9 and BMP9 + NGF respectively.The expression level of COL1,RUNX2 was detected by RT-PCR and Western blot,ALP activity was examined by ALP kit 3,12,24,48 hours,3 and 7 days after treatment,respectively.Results:The ALP activity of BMP9 + NGF group was the highest among the 4 groups.The difference in the groups firstly appeared at 3 h after treatment.The highest expression level of RUNX2 and COL1 was detected in BMP9 + NGF group.Conclusion:NGF and BMP9 may synergisticly promote osteogenic differentiation at the early stage of osteogenic induction of C3H10T1/2 cells.

Clinical efficacy on drynaria injection for delayed healing of fracture patients / 中国生化药物杂志

Objective To investigate the clinical study on drynaria injection for delayed healing of fracture patients.Methods 108 patients with fracture delayed union from February 2010 to February 2016 in our hospital were selected and randomly divided into the control group and the experiment group by different treatment,54 cases in each group.The control group were treated by conventional therapy,the experiment group were treated with drynaria rhizome injection on the bases of control group.The total effective rate,serum alkaline phosphatase,NGF and BMP-2 levels were observed and compared after treatment.Results After treatment,the total effective rate of experiment group was higher than control group (P<0.05);After treatment for 7 and 14 days,the serum alkaline phosphatase level of the two groups were significantly increased (P<0.05),and compared with the control group, the serum alkaline phosphatase content of the experimental group was higher (P<0.05);After treatment for 7 and 14 days,the serum NGF and BMP-2 levels of two groups were significantly increased(P<0.05),and compared with the control group,the experimental group was higher than those in the control group.Conclusion Injection of rhizoma drynariae in the treatment of delayed healing of patients with significant effect,increase the levels of serum alkaline phosphatase, serum NGF and BMP-2,promote fracture healing in patients and improve clinical efficacy.

Memantine improves cognitive deficits by activiating NGF/TrkA signaling in APP/PS1 transgenic mice / 中国药理学通报

Aims To study the role of NGF/Trk A sig-naling pathway in Memantine ( MEM) improving APP/PS1 transgenic mice cognitive deficits and to explore its possible mechanisms. Methods Cognitive perform-ance was assessed by Morris water maze( MWM) , pas-sive avoidance test( PAT) and locomotivity test. Aβ1-42 protein levels were determined by immunohistochemis-try. The activities of AChE and ChAT were also exam-ined by ELISA and colorimetry. Western blot was used to detect the expression levels of NGF and its receptor TrkA and the downstream ERK pathway. Results MEM treatment significantly ameliorated the cognitive deficits, dramatically reduced the Aβ1-42 overexpres-sion. MEM increased the activity of choline acetyl-transferase( ChAT) , while decreased that of acetylcho-line esterase( AChE) . Moreover, MEM activiated NGF signaling by increasing the phosphorylation of TrkA fol-lowing the increased phosphorylation of c-Raf, ERK1/2 and downstream effector CREB after MEM treatment. Conclusion MEM treatment may activate the NGF/TrkA signaling in APP/PS1 mice to reduce amyloidosis and cognitive deficits.

Lentivirus-mediated NGF gene silencing inhibited differentiation of PC12 Cells / 中国药理学通报

Aim To investigate the effects of lentivirus mediated nerve growth factor ( NGF) gene silencing on pheochromocytoma cells ( PC12 ) and the possible mechanisms .Methods The NGF shRNA expression vector was constructed .PC12 cells were randomly divi-ded into five groups (n=3 each) as follows: negative control group ( NC ) , control lentivirus group ( LV CON) , lentivirus NGF shRNA1 group ( LV shNGF1 ) , lentivirus NGF shRNA2 group(LV shNGF2), lentivir-us NGF shRNA3 group(LV shNGF3).The cells in NC group were cultured in DMEM/HG and polybrene me-dium, while others were cultured in DMEM/HG, poly-brene and corresponding lentivirus medium .After the treatment, the infection efficiency was determined by fluorescent microscope .Relative expression of NGF , extracellular signal-regulated kinase ( ERK1/2 ) and p-ERK1/2 were assessed by Western blot .The expres-sion of NGF mRNA was analyzed by quantitative re-verse transcription polymerase chain reaction ( qRT-PCR) .The differentiation degree was valued according to the length of neuritis and max diameter of cells .The cell viability was detected by CCK-8.Results The in-fection efficiency in PC12 cells reached over 90%. Compared with NC group , the relative expression of NGF mRNA and NGF protein was significantly down-regulated ( P<0.05 ) .There was no difference in the expression of ERK1/2 protein and cell viability .The expression of p-ERK1/2 protein was markedly down-regulated in LV shNGF3 group ( P<0.01 ) .The cells morphology was changed , and the length of neuritis and max diameter of cells were strained in LV shNGF 3 group than those in NC group ( P<0.01 ) .Conclusion Lentivirus-mediated NGF gene silencing inhibits the differentiation of PC12 cells through suppressing the activation of ERK1/2.

Efficacy and Influence of Rat NGF on Serum Neuropeptide Y and Neuron-specificenolase Levels in New-born with Neonatal Hypoxic and Ischemic Encephalopathy / 中国药师

To discuss the therapeutic effect and influence of rat NGF on serum neuropeptide Y ( NPY) and neuron-specificenolase (NSE) levels in newborn with neonatal hypoxic and ischemic encephalopathy (HIE). Methods: Totally 70 cases of neonatal HIE patients were selected and divided into the observation group and the control group at random. The children in the two groups were given the routine medical treatment such as oxygen uptake, the control of intracranial pressure, blood pressure and blood sugar, anticonvulsant, balance of water and electrolyte and etc. The children in the observation group were additionally given 20μg rat NGF through intramuscular injection once a day. The children in the control group were additionally given 100 mg citicoline through in-travenous drip once a day. The treatment course was 10-14 days. The changes of serum NPY and NSE levels, clinical curative effect and adverse drug reactions ( ADR) in the two groups were compared, and the occurrence of nervous system sequel after the medical treatment was compared and followed up. Results: After the 2-week medical treatment, the serum NPY and NSE levels in the two groups were declined (P<0. 05 or P<0. 01), and the declining in the observation group was more notable than that in the control group (P<0. 05). The total clinical efficiency in the observation group was 94. 28%, which was much higher than that (68. 57%) in the control group (P<0. 01). No obvious adverse drug reaction appeared in the two groups during the treatment. After the one-year follow up, the occurrence of sequel was 5. 71% in the observation group, which was much lower than that (22. 86%) in the control group(P<0. 05). Conclusion:Rat NGF has reliable curative effect on neonatal HIE with high security, which can improve the repair of impaired neuronal cells, reduce the occurrence of sequel of neuronal cells and enhance the prognosis, and the mechanism may relat-ed with the reduction of serum NPY and NSE levels.