脑源性神经生长因子抗体对小鼠坐骨神经损伤后脊髓与背根节内Synaptophysin表达的影响

目的探查小鼠坐骨神经压榨损伤后内源性脑源性神经营养因子(brainderivedneuroliophicfactor,BDNF)对Synaptophysin(SYN)在相应脊髓前角运动细胞与背根节神经元内表达的影响。方法在小鼠一侧坐骨神经压榨损伤后腹腔注射BDNF抗体,中和内源性BDNF,然后用免疫组织化学方法观察SYN在与坐骨神经相连的脊髓前角运动细胞与背根节神经元的表达。结果实验组SYN免疫反应阳性神经元的数目较对照组明显减少,阳性细胞的平均光密度也显著下降(P<0.01)。结论小鼠坐骨神经压榨损伤后内源性BDNF可能参与脊髓前角运动细胞与背根节神经元内SYN的表达。

猴垂体前叶Synaptophysin免疫反应性的光镜观察

目的：为神经纤维对哺乳动物尤其是灵长类动物垂体前叶腺细胞的直接调节作用提供形态学证据．方法：用抗Ｓｙｎａｐｔｏｐｈｙｓｉｎ（ＳＹＮ）单克隆抗体，对ＭａｃａｃａＭｕｌａｔａ猴垂体前叶进行了免疫组化染色．结果：猴垂体前叶ＳＹＮ的免疫反应性主要有膨体型和纤维型两种形式．膨体型表现为大小不等、深染的点状结构．纤维型表现为细、短浅染的ＳＹＮ免疫反应神经纤维，其上分布着大小不等、深染的膨体．ＳＹＮ免疫反应膨体和纤维均分布在腺细胞之间，或沿着细胞团边界走行，与腺细胞密切接触．结论：猴垂体前叶ＳＹＮ免疫反应膨体反映了神经末梢中突触小泡的存在，为“哺乳动物垂体前叶神经体液双重调节的假说”

Acupuncture accelerates neural regeneration and synaptophysin production after neural stem cells transplantation in mice

BACKGROUND Synaptophysin plays a key role in synaptic development and plasticity of neurons and is closely related to the cognitive process of Alzheimer’s disease(AD)patients.Exogenous neural stem cells(NSCs)improve the damaged nerve function.The effects of Sanjiao acupuncture on cognitive impairment may be related to the regulation of the NSC microenvironment.AIM To explore the anti-dementia mechanism of acupuncture by regulating the NSC microenvironment.METHODS NSCs were isolated from pregnant senescence-accelerated mouse resistant 1(SAMR1)mice,labeled with BrdU,and injected into the hippocampus of senescence-accelerated mouse prone 8(SAMP8)mice.Eight-month-old senescence-accelerated mice(SAM)were randomly divided into six groups:SAMR1(RC),SAMP8(PC),sham transplantation(PS),NSC transplantation(PT),NSC transplantation with acupuncture(PTA),and NSC transplantation with nonacupoint acupuncture(PTN).Morris water maze test was used to study the learning and memory ability of mice after NSC transplantation.Hematoxylin-eosin staining and immunofluorescence were used to observe the histopathological changes and NSC proliferation in mice.A co-culture model of hippocampal slices and NSCs was established in vitro,and the synaptophysin expression in the hippocampal microenvironment of mice was observed by flow cytometry after acupuncture treatment.RESULTS Morris water maze test showed significant cognitive impairment of learning and memory in 8-mo-old SAMP8,which improved in all the NSC transplantation groups.The behavioral change in the PTA group was stronger than those in the other two groups(P<0.05).Histopathologically,the hippocampal structure was clear,the cell arrangement was dense and orderly,and the necrosis of cells in CA1 and CA3 areas was significantly reduced in the PTA group when compared with the PC group.The BrdU-positive proliferating cells were found in NSC hippocampal transplantation groups,and the number increased significantly in the PTA group than in the PT and PTN groups(P<0.05).Flow cytometry showed that after co-culture of NSCs with hippocampal slices in vitro,the synaptophysin expression in the PC group decreased in comparison to the RC group,that in PT,PTA,and PTN groups increased as compared to the PC group,and that in the PTA group increased significantly as compared to the PTN group with acupointrelated specificity(P<0.05).CONCLUSION Acupuncture may promote nerve regeneration and synaptogenesis in SAMP8 mice by regulating the microenvironment of NSC transplantation to improve the nerve activity and promote the recovery of AD-damaged cells.

Ca^(2+)诱导的synaptophysin Ⅰ蛋白的脂筏分布

文章研究了Ca2+对synaptophysin Ⅰ(Syp Ⅰ)蛋白的脂筏分布的影响。研究结果证明,Syp Ⅰ蛋白的脂筏分布明显受到Ca2+的特异性调控。在无Ca2+的条件下,Syp Ⅰ为典型的非脂筏蛋白;而在低浓度Ca2+的条件下,Syp Ⅰ可以转变为脂筏结合蛋白。文章还研究了SypⅠ在Ca2+的诱导下进入脂筏膜微区的分子机制。研究结果表明,Syp Ⅰ在Ca2+的诱导下进入脂筏这一现象依赖于其C末端胞质区,确定了Syp Ⅰ的胞质区在这种调节中的重要性。

Effects of ephedrine on expression of Nogo-A and synaptophysin in neonatal rats following hypoxic-ischemic brain damage

BACKGROUND: Central nervous system axons regenerate poorly following neonatal hypoxic-ischemic brain damage (HIBD), partly due to inhibitors, such as Nogo-A. Very few studies have addressed the regulation of Nogo-A in neonatal rats following HIBD. However, numerous studies have shown that ephedrine accelerates neuronal remodeling and promotes recovery of neural function in neonatal rats following HIBD. OBJECTIVE: To investigate the effects of ephedrine on expression of Nogo-A and synaptophysin in brain tissues of neonatal rats following HIBD. DESIGN, TIME AND SETTING: A completely randomized, controlled study was performed at the Immunohistochemistry Laboratory of the Research Institute of Pediatrics, Children's Hospital of Chongqing Medical University from August 2008 to March 2009. MATERIALS: Ephedrine hydrochloride (Chifeng Pharmaceutical Group, China), rabbit anti-Nogo-A polyclonal antibody (Abcam, UK), and rabbit antisynaptophysin polyclonal antibody (Lab Vision, USA) were used in this study. METHODS: A total of 96 healthy, neonatal, Sprague Dawley rats were randomly assigned to three groups (n = 32): sham operation, HIBD, and ephedrine. The HIBD model was established by permanent occlusion of the left common carotid artery, followed by 2 hours of hypoxia (8% oxygen and 92% nitrogen). In the sham operation group, the left common carotid artery was exposed, but was not ligated or subjected to hypoxia. Rats in the ephedrine group were intraperitoneally injected with ephedrine immediately following HIBD, with 1.5 mg/kg each time. Rats in the sham operation and HIBD groups were injected with an equal volume of saline. All neonatal rats were treated once daily for 7 days. MAIN OUTCOME MEASURES: Histopathological damage to the cortex and hippocampus was determined by hematoxylin-eosin staining. Expression of Nogo-A and synaptophysin was detected using immunohistochemical staining. RESULTS: Neuronal degeneration and edema were observed in the hypoxic-ischemic cortex and hippocampus by hematoxylin-eosin staining. Compared with the sham operation group, the levels of Nogo-A significantly increased in the HIBD group at various time points (P < 0.01). Nogo-A expression was significantly reduced in the ephedrine group compared with the HIBD group (P < 0.01). Synaptophysin expression was significantly decreased in the hypoxic-ischemic cortex, compared with the sham operation group (P < 0.01). Synaptophysin levels were significantly increased in the ephedrine group, compared with the HIBD group (P < 0.01). CONCLUSION: Altered Nogo-A expression was associated with inversely altered synaptophysin expression. The use of ephedrine normalized expression levels of Nogo-A and synaptophysin following HIBD.

Effects of “Nourishing Liver and Kidney” Acupuncture Therapy on Expression of Brain Derived Neurotrophic Factor and Synaptophysin after Cerebral Ischemia Reperfusion in Rats

The aim of the present study was to investigate the effect of "nourishing liver and kidney" acupuncture therapy on motor and cognitive deficits,and the underlying mechanism following cerebral ischemia-reperfusion(I/R) via increasing the expression of brain derived neurotrophic factor(BDNF) and synaptophysin(SYN) in the hippocampus.Healthy adult male SD rats were randomly divided into sham operation group(n=51),model group(n=51),acupuncture group(n=51) and acupuncture control group(n=51).The middle cerebral I/R model was established.Acupunctures were performed in the acupuncture group and acupuncture control group at acupoints of Taixi(K103),Taichong(ST09) of both sides,for 30 min once daily every morning.The animals in the sham operation group and model group were conventionally fed in the cage,without any intervention therapy.The rats of each group were assessed with modified neurological severity scores(m NSS).The expression of BDNF and SYN in the hippocampus was detected by immunohistochemical SP method and the synaptic structure in hippocampus area was assessed morphologically and quantitatively at the 3rd,7th and 14 th day.The Morris water Maze(MWM) test was used to evaluate the rats' learning and memory abilities on the 15 th day after acupuncture.The animals in the acupuncture control group and sham operation group presented no neurological deficit.In the acupuncture group,the nerve functional recovery was significantly better than that in the model group at the 7th and 14 th day after modeling.The average MWM escape latency in the acupuncture group was shorter than that in the model group at the 3rd,4th and 5th day.The number of crossings of the platform quadrant in the acupuncture group was significantly more than that in the model group.At the each time point,the expression levels of BDNF and SYN in the hippocampal regions increased significantly in the model group as compared with the sham operation group and the acupuncture control group.In the acupuncture group,the expression levels of BDNF at the 7th and 14 th day increased more significantly than those in the model group.In the acupuncture group,the expression levels of SYN at the each time point increased more significantly than those in the model group.The post-synaptic density(PSD) was significantly increased and the synapse cleft width was narrowed in the acupuncture group as compared with other groups.The synaptic curvatures were improved obviously in the acupuncture group in contrast to the model group.It was concluded that the "nourishing liver and kidney" acupuncture therapy has positive effects on behavioral recovery,as well as learning and memory abilities,probably by promoting the expression of BDNF and SYN,and synaptic structure reconstruction in the ipsilateral hippocampus after I/R in rats.The "nourishing liver and kidney" acupuncture therapy can promote the functional recovery in rats after cerebral ischemia injury.

Changes in Synapses and Axons Demonstrated by Synaptophysin Immunohistochemistry Following Spinal Cord Compression Trauma in the Rat and Mouse

Objective and methods To evaluate synaptic changes using synaptophysin immunohistochemstry in rat and mouse, which spinal cords were subjected to graded compression trauma at the level of Th8-9. Results Normal animals showed numerous fine dots of synaptophysin immunoreactivity in the gray matter. An increase in synaptophysin immunoreactivity was observed in the neuropil and synapses at the surface of motor neurons of the anterior horns in the ThS-9 segments lost immunoreactivity at 4-hour point after trauma. The immunoreactive synapses reappeared around motor neurons at 9-day point. Unexpected accumulation of synaptophysin immunoreactivity occurred in injured axons of the white matter of the compressed spinal cord. Conclusion Synaptic changes were important components of secondary injuries in spinal cord trauma. Loss of synapses on motor neurons may be one of the factors causing motor dysfunction of hind limbs and formation of new synapses may play an import,ant role in recovery of motor function. Synaptophysin immunohistochemistry is also a good tool for studies of axonal swellings in spinal cord injuries.

Influence of acupuncture with exercise training on learning and memory functions, as well as microtubule-associated protein-2 and synaptophysin expression in the hippocampal CA3 region, in a rat model of cerebral infarction

The present study was designed to determine microtubule-associated protein-2 and synaptophysin expression in the hippocampal CA3 region in a rat model of middle cerebral artery occlusion. The rats were treated with acupuncture at Baihui (GV 20), Qubin (GB 7), and Qianding (GV 21) points, in addition to exercise training. Results were compared with rats undergoing exercise training only. The Y-maze method and immunohistochemistry revealed decreased error frequency of passing through Y-maze, as well as significantly increased microtubule-associated protein-2 and synaptophysin expression, in the acupuncture with exercise training group compared with the model and exercise training groups after 5 weeks. Microtubule-associated protein-2 and synaptophysin expressions negatively correlated with error frequency of passing through the Y-maze. These results suggested that acupuncture combined with exercise training improved learning and memory functions in a rat model of cerebral infarction. The mechanisms of action were hypothesized to be associated with dendritic or synaptic plasticity in the ipsilateral hippocampal CA3 region.

Combined acupuncture and HuangDiSan treatment affects behavior and synaptophysin levels in the hippocampus of senescence-accelerated mouse prone 8 after neural stem cell transplantation

Sanjiao acupuncture and Huang Di San can promote the proliferation, migration and differentiation of exogenous neural stem cells in senescence-accelerated mouse prone 8(SAMP8) mice and can improve learning and memory impairment and behavioral function in dementia-model mice. Thus, we sought to determine whether Sanjiao acupuncture and Huang Di San can elevate the effect of neural stem cell transplantation in Alzheimer's disease model mice. Sanjiao acupuncture was used to stimulate Danzhong(CV17), Zhongwan(CV12), Qihai(CV6), bilateral Xuehai(SP10) and bilateral Zusanli(ST36) 15 days before and after implantation of neural stem cells(5 × 105) into the hippocampal dentate gyrus of SAMP8 mice. Simultaneously, 0.2 m L Huang Di San, containing Rehmannia Root and Chinese Angelica, was intragastrically administered. Our results demonstrated that compared with mice undergoing neural stem cell transplantation alone, learning ability was significantly improved and synaptophysin m RNA and protein levels were greatly increased in the hippocampus of mice undergoing both Sanjiao acupuncture and intragastric administration of Huang Di San. We conclude that the combination of Sanjiao acupuncture and Huang Di San can effectively improve dementia symptoms in mice, and the mechanism of this action might be related to the regulation of synaptophysin expression.

Effect of Panaxtriol Saponins on synaptophysin and postsynaptic density-95 expression at different periods of cerebral infarction

BACKGROUND:The change in expression of synaptophysin (Syp) and postsynaptic density-95 (PSD-95) alters after cerebral infarction,and the plasticity of synapses contributes greatly to nerve function recovery. Chinese medicinal substances may play an important role in the expression of Syp and PSD-95. OBJECTIVE: To observe the effect of Panaxtriol Saponins (PTS),an active component in Sanqi tongshu capsules,on the expression of Syp and PSD-95 after cerebral infarction at different time points in rats,so as to examine the cerebral function remodeling mechanism. DESIGN,TIME AND SETTING: A randomized and controlled observation which was performed in Dongzhimen Hospital,Beijing University of Traditional Chinese Medicine from January to March,2007. MATERIALS: Twenty-six healthy male Sprague Dawley rats were used to establish middle cerebral artery occlusion based on the Longa method. Sanqi tongshu capsules (containing 100 mg PTS per tablet) were provided by the Chengdu Huashen Group and nimodipine tablets (30 mg) by Tianjin Zhongyang Pharmaceutical Co.,Ltd. METHODS: Twenty-six rats were randomly divided into an operation group (n=21) and a control group (n=5). The operation group underwent the EZ Longa procedure to make the middle cerebral artery occlusion model. After surgery rats were randomly divided into a model group,a PTS group and a nimodipine group,with seven rats in each group. Rats were intragastrically administrated with saline (2 mL/d) in the model group,with Sanqi tongshu capsule (5.4 mg/100 g/d) in the PTS group,and with nimodipine (1.73 mg/100 g/d) in the nimodipine group. Rats in the control group did not undergo model establishment and drug administration. MAIN OUTCOME MEASURES: The expressions of Syp and PSD-95 were measured by immunohistochemical and image analysis at days 3,7 and 28 after the operation. RESULTS: The expression of Syp and PSD-95 in the operation group was significantly lower than in the control group at days 3,7,28 postoperatively (P<0.05). The expression of Syp and PSD-95 in the PTS group and nimodipine group was significantly higher than in the model group at day 28 postoperatively (P< 0.05-0.01). Additionally,after PTS and nimodipine treatment at different intervals,the expression of Syp and PSD-95 at day 28 postoperatively was significantly higher than those at days 3 and 7 postoperatively,respectively (P<0.01). CONCLUSION: PTS can promote the expression of Syp and PSD-95,i.e. the remodeling process of synapses,after cerebral infarction at different time points in rats,which contributes to cerebral function remodeling.

Effects of genistein and 17 beta-estradiol on hippocampal synaptophysin expression in ovariectomized rats

BACKGROUND: Phytoestrogen,derived from plants,is an estrogen-like element,and is effective and safe for estrogen replacement. OBJECTIVE: To compare the interventional effects of genistein and 17β-estradiol on learning and mem-ory and synaptophysin (SYN) expression in the hippocampus of ovariectomized rats. DESIGN: Randomized controlled animal study. SETTING: Department of Neurology,the Third Affiliated Hospital,Xiangya Medical College,Central South University. MATERIALS: 130 healthy female Sprague Dawley (SD) rats,6 months old and weighing (293.1 ± 10.2) g,were provided by the Second Xiangya Hospital of Central South University. This animal experiment received confirmed consent from the local ethics committee. All rats were randomly divided into 5 groups,including baseline group (n = 10),sham operation group (n = 30),ovariectomized group (n = 30),genistein group (n = 30),and 17β-estradiol group (n = 30). Rats in the latter four groups were observed for 3 weeks (n = 10) and for 15 weeks (n = 20) after model establishment. METHODS: This study was performed at the Department of Endocrinology,the Second Affiliated Hos-pital,Xiangya Medical College,Central South University from August 2005 to January 2006. Animals were not submitted to any treatment in the baseline group,but anesthetized and sacrificed at the 7 months of age. After anesthesia in the ovariectomized,genistein,and 17β-estradiol groups,both ovaries were separated and resected to establish an ovariectomized model. The same volume of fat was resected in the sham operation group. After surgery,rats were intraperitoneally injected with 5 mg/kg genistein in the genistein group,10 μg/kg 17β-estradiol in the 17β-estradiol group,and 0.1 mL/100 g dimethyl sulfoxide (DMSO)/polyethylene glycol (PEG)-200 stock solution in the sham operation and ovariectomized groups once a day until one day before sacrifice. MAIN OUTCOME MEASURES: ① Learning and memory changes of SD rats were detected using water maze behavioral testing 3 and 15 weeks after surgery. ② SYN expression in the hippocampus was meas-ured using immunohistochemistry. RESULTS: A total of 16 out of 130 rats died due to infection,and 114 rats were included in the final analy-sis. ① Comparison of water maze results from the five groups: by 3 and 15 weeks after surgery,escape la-tency was prolonged and platform-crossing times decreased in the ovariectomized group compared to the baseline,genistein,17β-estradiol,and sham operation groups (t = 4.17-14.64,P < 0.05). However,there were no significant differences in escape latency and platform-crossing times among the sham operation,genistein,and 17β-estradiol groups (P > 0.05). ② Distribution and quantity of SYN immunoreactive products in hippocampus: SYN-immunoreactive cells stained darkly in the baseline and sham operation groups,but were lightly stained in the genistein,17β-estradiol,and ovariectomized groups. In particular,SYN-immunoreactive cells stained lightly in the ovariectomized group 15 weeks after surgery. SYN correc-tion gray values in hippocampal sub-regions,especially in the mossy fiber layer of the CA3 region,of the ovariectomized group was lower compared to the baseline,sham operation,17β-estradiol,and genistein groups (t = 12.57-23.92,P < 0.05) 15 weeks after surgery. However,there were no significant differences in SYN correction gray values among the baseline,sham operation,17β-estradiol and genistein groups (P > 0.05). CONCLUSION: Genistein or 17β-estradiol supplemental therapy antagonizes memory deterioration,due to endogenous estrogen deficiency and blocks the decrease of SYN expression in the hippocampus. The ef-fect of genistein is similar to 17β-estradiol.

Effects of electroacupuncture versus nimodipine on long-term potentiation and synaptophysin expression in a rat model of vascular dementia

The present study stimulated Baihui (DU 20) and Dazhui (DU 14) acupoints in a rat model of vascular dementia with electroacupuncture to investigate changes in long-term potentiation and synaptophysin expression in the hippocampus.The results revealed that synaptophysin expression in brain tissues was increased after electroacupuncture.After high-frequency stimulation,the population spike latency was shortened and the excitatory postsynaptic potential slope and population spike amplitude were increased.In addition,cognitive function was enhanced,similar to the effects of intragastric perfusion of nimodipine.The results indicated that electroacupuncture at Baihui and Dazhui acupoints can improve learning and memory functions of a rat model of vascular dementia by promoting synaptophysin expression,enhancing hippocampal synaptic plasticity and accelerating synaptic transmission.

Perlecan and synaptophysin changes in denervated skeletal muscle

The present study observed sciatic nerve and gastrocnemius muscle changes in denervated rats using morphology methods,and assessed expression of perlecan,an extracellular matrix com-ponent,which is located at the skeletal muscle cell surface as acetylcholine esterase,as well as synaptophysin,a synaptic marker.Results showed degeneration and inflammation following transection of the sciatic nerve.In addition,the sciatic nerve-dominated skeletal muscle degen-erated with mild inflammation,indicating that skeletal muscle atrophy primarily contributed to denervation-induced nutritional disturbances.With prolonged injury time(1-4 weeks post-injury),perlecan expression gradually decreased and reached the lowest level at 4 weeks,but synap-tophysin expression remained unchanged after denervation.Results suggested that perlecan expression was more sensitive to denervation and reflected regional extracellular matrix changes following denervation.

人参皂苷Rh2对大鼠C6胶质瘤细胞Siah-1、Synaptophysin、MMP9及VEGF表达的影响

目的:探讨人参皂苷Rh2对大鼠C6胶质瘤细胞Siah-1、突触素(Synaptophysin)、基质金属蛋白酶-9(MMP-9)血管内皮生长因子(VEGF)表达的影响。方法:将大鼠C6胶质瘤细胞分为对照组、人参皂苷Rh2低剂量组(16μg/m L)、人参皂苷Rh2中剂量组(32μg/m L)、人参皂苷Rh2高剂量组(48μg/m L),CCK-8法和平板克隆实验检测细胞增殖;流式细胞术检测细胞凋亡;Transwell检测细胞侵袭;实时荧光定量-聚合酶链式反应(qRT-PCR)检测C6胶质瘤细胞中VEGF、Siah-1、Synaptophysin、MMP-9 m RNA表达;蛋白质印迹法(Western blot)检测C6胶质瘤细胞中VEGF、Siah-1、Synaptophysin、MMP-9蛋白表达。结果:与对照组比较,人参皂苷Rh2低剂量组、人参皂苷Rh2中剂量组、人参皂苷Rh2高剂量组大鼠C6胶质瘤细胞OD_(450)值(24 h、48 h)、克隆形成率、细胞侵袭数、VEGF、Synaptophysin、MMP-9 m RNA及蛋白表达降低,细胞凋亡率、Siah-1 m RNA及蛋白表达升高,且呈剂量依赖性(P<0.05)。结论:人参皂苷Rh2可能通过上调Siah-1,下调VEGF、Synaptophysin、MMP-9表达来抑制大鼠C6胶质瘤细胞增殖与侵袭,促进细胞凋亡。

The effects of bone marrow mesenchymal stromal cells transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA_3 region in hippocampus of vascular dementia rats

Objective To investigate the effects of bone marrow mesenchymal stromal cells (BMSCs) transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3region in hippocampus of vascular dementia (VD) rats.Methods

Repetitive transcranial magnetic stimulation promotes neurological functional recovery in rats with traumatic brain injury by upregulating synaptic plasticity-related proteins

Studies have shown that repetitive transcra nial magnetic stimulation(rTMS)can enhance synaptic plasticity and improve neurological dysfunction.Howeve r,the mechanism through which rTMS can improve moderate traumatic brain injury remains poorly understood.In this study,we established rat models of moderate traumatic brain injury using Feeney's weight-dropping method and treated them using rTMS.To help determine the mechanism of action,we measured levels of seve ral impo rtant brain activity-related proteins and their mRNA.On the injured side of the brain,we found that rTMS increased the protein levels and mRNA expression of brain-derived neurotrophic factor,tropomyosin receptor kinase B,N-methyl-D-aspartic acid receptor 1,and phosphorylated cAMP response element binding protein,which are closely associated with the occurrence of long-term potentiation.rTMS also partially reve rsed the loss of synaptophysin after injury and promoted the remodeling of synaptic ultrastructure.These findings suggest that upregulation of synaptic plasticity-related protein expression is the mechanism through which rTMS promotes neurological function recovery after moderate traumatic brain injury.

Chronic Kidney Disease Induces Cognitive Impairment in the Early Stage

Objective Previous research indicates a link between cognitive impairment and chronic kidney disease(CKD),but the underlying factors are not fully understood.This study aimed to investigate the progression of CKD-induced cognitive impairment and the involvement of cognition-related proteins by developing early-and late-stage CKD models in Sprague-Dawley rats.Methods The Morris water maze test and the step-down passive avoidance task were performed to evaluate the cognitive abilities of the rats at 24 weeks after surgery.Histopathologic examinations were conducted to examine renal and hippocampal damage.Real-time PCR,Western blotting analysis,and immunohistochemical staining were carried out to determine the hippocampal expression of brain-derived neurotrophic factor(BDNF),choline acetyltransferase(ChAT),and synaptophysin(SYP).Results Compared with the control rats,the rats with early-stage CKD exhibited mild renal damage,while those with late-stage CKD showed significantly increased serum creatinine levels as well as apparent renal and brain damage.The rats with early-stage CKD also demonstrated significantly impaired learning abilities and memory compared with the control rats,with further deterioration observed in the rats with late-stage CKD.Additionally,we observed a significant downregulation of cognition-related proteins in the hippocampus of rats with early-stage CKD,which was further exacerbated with declining renal function as well as worsening brain and renal damage in rats with late-stage CKD.Conclusion These results suggest the importance of early screening to identify CKD-induced cognitive dysfunction promptly.In addition,the downregulation of cognition-related proteins may play a role in the progression of cognitive dysfunction.

藏药十八味诃子利尿丸对雄性糖尿病大鼠脑组织离子钙结合衔接分子1及突触可塑性相关蛋白的影响

目的探讨藏药十八味诃子利尿丸对雄性糖尿病(Diabetes mellitus,DM)大鼠脑组织中小胶质细胞标记物离子钙结合衔接分子1(Ionized calcium binding adaptor molecule 1,IBA1)及突触可塑性相关蛋白的影响。方法30只雄性SD大鼠适应性喂养72 h后,取10只大鼠为对照组;另取20只制备DM大鼠,以高糖高脂饲料喂养12 w后于尾静脉一次性注射STZ溶液,其中10只为模型组、10只为藏药组(藏药组用十八味诃子利尿丸连续灌胃2 w),实验结束后断头取脑组织。用Western Blot法检测IBA1、IL-6、syn-1、PSD95、HIF-1α,用IHC法检测IBA1、IL-6、HIF-1α。对实验结果进行相关统计学分析。结果与对照组比较,模型组大鼠脑组织中的IBA1、IL-6、HIF-1α蛋白表达升高(P<0.05),syn-1、PSD95蛋白表达降低(P<0.05);与模型组比较,藏药组大鼠脑组织中的IBA1、IL-6、HIF-1α蛋白表达降低(P<0.05),syn-1、PSD95蛋白表达量升高(P<0.05)。结论藏药十八味诃子利尿丸可能通过IBA1靶点降低DM大鼠脑组织中小胶质细胞的活化水平,并改善突触可塑性,从而对DM大鼠脑组织发挥保护作用。

NaAsO_(2)通过Hippo通路影响PC12细胞形态和突触蛋白

砷是一种自然环境中广泛存在的非金属元素[1]。人类在自然环境和工业活动中长期接触受砷污染的水、空气和食物而中毒,造成组织器官损伤、甚至癌变。此外,砷引起神经系统损害也受到人们越来越多的关注[2]。但目前砷引起神经损伤的具体机制仍不是十分明确。Hippo信号通路是一条在进化上高度保守的激酶级联信号通路,主要控制器官大小、组织稳态和组织再生[3]。本课题组已经证实,NaAsO_(2)可通过激活Hippo信号通路,诱导PC12细胞凋亡。突触后致密蛋白95(post synaptic density protein,PSD95)、突触素蛋白(synaptophysin,SYN)作为神经突触功能相关蛋白,其表达和缺失在神经系统疾病中十分重要[4]。因此,本实验在前期研究的基础上探讨Hippo通路是否参与NaAsO_(2)对PC12细胞活性、形态以及PSD95、SYN神经突触相关蛋白表达的影响。

锁阳乙酸乙酯提取物的雌激素样作用研究

雌激素提高学习记忆能力的功能已被广泛承认,前期实验证实锁阳乙酸乙酯提取物(ECS)可表现出雌激素样作用,提高去卵巢大鼠的学习记忆能力及血清E2水平,对Aβ损伤SK-N-SH细胞有一定的保护作用。为了进一步研究ECS的雌激素样作用机制,拟在小鼠神经母细胞瘤细胞(Neuro2A)上探寻ECS对学习记忆相关蛋白突触素蛋白synaptophysin和环磷腺苷效应元件结合蛋白表达影响。采用CCK8法测定ECS对小鼠神经母细胞瘤细胞(Neuro2A)细胞活力的影响。将小鼠神经母细胞瘤细胞分为正常组、DMSO组、ECS组、Tamoxifen组、Tamoxifen+ECS组;正常组、ECS组、ECS+G15组和正常组、DMSO组、ECS组、PD98059组、PD98059+ECS组。采用Western blot法测定synaptophysin,p-creb蛋白表达情况。实验表明锁阳乙酸乙酯提取物可促进synaptophysin,P-CREB表达,ER受体竞争性拮抗剂Tamoxifen可降低synaptophysin表达,对ECS促进synaptophysin表达无显著影响。GPR30受体特异性阻断剂G15可显著降低ECS促进synaptophysin表达。p-CREB上游Erk1/2-MAPK通路的阻断剂PD98059可显著抑制ECS上调p-CREB表达,结合前期实验结果,我们认为ECS可与GPR30结合通过MAPK通路促进学习记忆相关蛋白synaptophysin表达。