AIM:To investigate the colocalization,density and profile of neuronal areas of enteric neurons in the ileum of male obese mice.METHODS:The small intestinal samples of male mice in an obese group(OG)(C57BL/6J ob/ob)and...AIM:To investigate the colocalization,density and profile of neuronal areas of enteric neurons in the ileum of male obese mice.METHODS:The small intestinal samples of male mice in an obese group(OG)(C57BL/6J ob/ob)and a control group(CG)(+/+)were used.The tissues were analyzed using a double immunostaining technique for immunoreactivity(ir)of the P2X2 receptor,nitric oxide synthase(NOS),choline acetyl transferase(ChAT)and calretinin(Calr).Also,we investigated the density and profile of neuronal areas of the NOS-,ChAT-and Calrir neurons in the myenteric plexus.Myenteric neurons were labeled using an NADH-diaphorase histochemical staining method.RESULTS:The analysis demonstrated that the P2X2receptor was expressed in the cytoplasm and in the nuclear and cytoplasmic membranes only in the CG.Neuronal density values(neuron/cm2)decreased 31%(CG:6579±837;OG:4556±407)and 16.5%(CG:7796±528;OG:6513±610)in the NOS-ir and calretininir neurons in the OG,respectively(P<0.05).Density of ChAT-ir(CG:6200±310;OG:8125±749)neurons significantly increased 31%in the OG(P<0.05).Neuron size studies demonstrated that NOS,ChAT,and Calr-ir neurons did not differ significantly between the CG and OG groups.The examination of NADH-diaphorase-positive myenteric neurons revealed an overall similarity between the OG and CG.CONCLUSION:Obesity may exert its effects by promoting a decrease in P2X2 receptor expression and modifications in the density of the NOS-ir,ChAT-ir and CalR-ir myenteric neurons.展开更多
AIM:To examine whether the ob/ob mouse model of obesity is accompanied by enteric nervous system ab-normalities such as altered motility METHODS:The study examined the distribution of the P2X 2 receptor (P2X 2 R) in m...AIM:To examine whether the ob/ob mouse model of obesity is accompanied by enteric nervous system ab-normalities such as altered motility METHODS:The study examined the distribution of the P2X 2 receptor (P2X 2 R) in myenteric neurons of female ob/ob mice. Specifically, we used immunohistochemistry to analyze the co-expression of the P2X 2 R with neuronal nitric oxide synthase (nNOS), choline acetyltrans-ferase (ChAT), and calretinin (CalR) in neurons of the small intestine myenteric plexus in ob/ob and control female mice In these sections, we used scanning confocal microscopy to analyze the co-localization of these markers as well as the neuronal density (cm 2 ) and area profile (μm2) of P2X 2 R-positive neurons In addition, enteric neurons were labeled using the nicotinamide adenine dinucleotide (NA H) diaphorase method and analyzed with light microscopy as an alternate means by which to analyze neuronal density and areaRESULTS:In the present study, we observed a 29 6% increase in the body weight of the ob/ob animals (OG) compared to the control group (CG) In addition, the average small intestine area was increased by approxi-mately 29 6% in the OG compared to the CG Immu-noreactivity (IR) for the P2X 2 R, nNOS, ChAT and CalR was detectable in the myenteric plexus, as well as in the smooth muscle, in both groups This IR appeared to be mainly cytoplasmic and was also associated with the cell membrane of the myenteric plexus neurons, where it outlined the neuronal cell bodies and their processes P2X 2 R-IR was observed to co-localize 100% with that for nNOS, ChAT and CalR in neurons of both groups In the ob/ob group, however, we observed that the neuronal density (neuron/cm 2 ) of P2X 2 R-IR cells was in-creased by 62% compared to CG, while that of NOS-IR and ChAT-IR neurons was reduced by 49% and 57%, respectively, compared to control mice The neuronal density of CalR-IR neurons was not different between the groups Morphometric studies further demonstrated that the cell body profile area (μm2) of nNOS-IR, ChAT-IR and CalR-IR neurons was increased by 34%, 20% and 55%, respectively, in the OG compared to controls Staining for NA H diaphorase activity is widely used to detect alterations in the enteric nervous system; however, our qualitative examination of NA H-diaphorase positive neurons in the myenteric ganglia revealed an overall similarity between the two groups CONCLUSION:We demonstrate increases in P2X2R expression and alterations in nNOS, ChAT and CalR IR in ileal myenteric neurons of female ob/ob mice compared to wild-type controls.展开更多
目的通过研究匹罗卡品致癫痫大鼠海马组织中P2X2受体(嘌呤受体)的表达变化,探讨其在颞叶癫痫发病中的作用机制。方法应用氯化锂-匹罗卡品腹腔注射建立大鼠颞叶癫痫模型,并予以P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)腹腔注射,应用We...目的通过研究匹罗卡品致癫痫大鼠海马组织中P2X2受体(嘌呤受体)的表达变化,探讨其在颞叶癫痫发病中的作用机制。方法应用氯化锂-匹罗卡品腹腔注射建立大鼠颞叶癫痫模型,并予以P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)腹腔注射,应用Western blot及Real-time PCR技术检测大鼠海马组织P2X2受体的表达,免疫组化技术检测各组大鼠海马组织中谷氨酸(glutamate,GLU)表达水平。结果 Western blot及Real-time PCR结果显示:慢性自发性癫痫发作组中P2X2表达明显增高(P<0.05),BBG干预组表达减低P<0.05)。免疫组化检测显示:GLU在BBG干预组较慢性自发性发作组中释放减少(P<0.05),且与BBG剂量呈负相关性。结论 P2X2受体通路可能参与颞叶癫痫的发病过程,并有望成为新一代治疗颞叶癫痫的药物靶点。展开更多
目的通过研究P2X2受体(嘌呤受体)蛋白在颞叶癫痫大鼠脑组织中的表达水平及星形胶质细胞的增殖水平,以及应用P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)后对颞叶癫痫的影响,探讨P2X2受体在颞叶癫痫发病机制中的作用。方法应用氯化锂-匹...目的通过研究P2X2受体(嘌呤受体)蛋白在颞叶癫痫大鼠脑组织中的表达水平及星形胶质细胞的增殖水平,以及应用P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)后对颞叶癫痫的影响,探讨P2X2受体在颞叶癫痫发病机制中的作用。方法应用氯化锂-匹罗卡品腹腔注射建立SD大鼠颞叶癫痫模型,将大鼠分为正常盐水对照组、急性发作组、慢性自发性发作组;其中慢性自发性发作组又分为空白对照组(生理盐水)和亮蓝G干预组,应用免疫组化检测各组大鼠海马区P2X2受体及胶质纤维酸性蛋白(GFAP)的表达水平,GFAP反映星形胶质细胞的增殖水平。结果免疫组化检测发现,P2X2受体主要表达于大鼠海马组织的CA1、CA3、齿状回、门区,并且在丘脑也有一定的表达;P2X2受体及GFAP在慢性自发性发作空白对照组中的表达明显高于正常盐水对照组和急性发作组(P<0.05);而亮蓝G干预组与慢性自发性发作空白对照组相比,P2X2受体及GFAP表达水平明显降低(P<0.05)。结论 P2X2受体可能在颞叶癫痫的发病机制中有重要作用,阻滞P2X2受体可能是治疗颞叶癫痫的潜在靶点。展开更多
Neural stem/progenitor cells：Radial glial cells constitute multipotent cells in the ventricular zone,lining the wall of the lateral ventricle of the embryonic brain.They have the capacity to give rise to cells belong...Neural stem/progenitor cells：Radial glial cells constitute multipotent cells in the ventricular zone,lining the wall of the lateral ventricle of the embryonic brain.They have the capacity to give rise to cells belonging to all three major linages（neurons,astrocytes and oligodendrocytes）of the nervous system（Tang and Illes,2017）.展开更多
Electroacupuncture at Shangjuxu （ST37） and Tianshu （ST25） can improve visceral hypersensitivity in rats. Colorectal distension was used to establish a rat model of chronic visceral hypersensitivity. Immunohistoche...Electroacupuncture at Shangjuxu （ST37） and Tianshu （ST25） can improve visceral hypersensitivity in rats. Colorectal distension was used to establish a rat model of chronic visceral hypersensitivity. Immunohistochemistry was used to detect P2X2 and P2X3 receptor expression in dorsal root ganglia from rats with chronic visceral hypersensitivity. Results demonstrated that abdominal withdrawal reflex scores obviously increased following establishment of the model, indicating visceral hypersensitivity. Simultaneously, P2X2 and P2X3 receptor expression increased in dorsal root ganglia. After bilateral electroacupuncture at Shangjuxu and Tianshu, abdominal withdrawal reflex scores and P2X2 and P2X3 receptor expression decreased in rats with visceral hypersensitivity. These results indicated that electroacupuncture treatment improved visceral hypersensitivity in rats with irritable bowel syndrome by reducing P2X2 and P2X3 receptor expression in dorsal root ganglia.展开更多
Painful diabetic neuropathy(PDN)is a diabetes mellitus complication.Unfortunately,the mechanisms underlying PDN are still poorly understood.Adenosine triphosphate(ATP)-gated P2X7 receptor(P2X7R)plays a pivotal role in...Painful diabetic neuropathy(PDN)is a diabetes mellitus complication.Unfortunately,the mechanisms underlying PDN are still poorly understood.Adenosine triphosphate(ATP)-gated P2X7 receptor(P2X7R)plays a pivotal role in non-diabetic neuropathic pain,but little is known about its effects on streptozotocin(STZ)-induced peripheral neuropathy.Here,we explored whether spinal cord P2X7R was correlated with the generation of mechanical allodynia(MA)in STZ-induced type 1 diabetic neuropathy in mice.MA was assessed by measuring paw withdrawal thresholds and western blotting.Immunohistochemistry was applied to analyze the protein expression levels and localization of P2X7R.STZ-induced mice expressed increased P2X7R in the dorsal horn of the lumbar spinal cord during MA.Mice injected intrathecally with a selective antagonist of P2X7R and P2X7R knockout(KO)mice both presented attenuated progression of MA.Double-immunofluorescent labeling demonstrated that P2X7R-positive cells were mostly co-expressed with Iba1(a microglia marker).Our results suggest that P2X7R plays an important role in the development of MA and could be used as a cellular target for treating PDN.展开更多
Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplante...Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells（OECs） remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.展开更多
基金Supported by Sao Paulo Research Foundation(FAPESP/Sao Paulo Research Foundation/Proc:05/04752-0)and CAPES Fellowship
文摘AIM:To investigate the colocalization,density and profile of neuronal areas of enteric neurons in the ileum of male obese mice.METHODS:The small intestinal samples of male mice in an obese group(OG)(C57BL/6J ob/ob)and a control group(CG)(+/+)were used.The tissues were analyzed using a double immunostaining technique for immunoreactivity(ir)of the P2X2 receptor,nitric oxide synthase(NOS),choline acetyl transferase(ChAT)and calretinin(Calr).Also,we investigated the density and profile of neuronal areas of the NOS-,ChAT-and Calrir neurons in the myenteric plexus.Myenteric neurons were labeled using an NADH-diaphorase histochemical staining method.RESULTS:The analysis demonstrated that the P2X2receptor was expressed in the cytoplasm and in the nuclear and cytoplasmic membranes only in the CG.Neuronal density values(neuron/cm2)decreased 31%(CG:6579±837;OG:4556±407)and 16.5%(CG:7796±528;OG:6513±610)in the NOS-ir and calretininir neurons in the OG,respectively(P<0.05).Density of ChAT-ir(CG:6200±310;OG:8125±749)neurons significantly increased 31%in the OG(P<0.05).Neuron size studies demonstrated that NOS,ChAT,and Calr-ir neurons did not differ significantly between the CG and OG groups.The examination of NADH-diaphorase-positive myenteric neurons revealed an overall similarity between the OG and CG.CONCLUSION:Obesity may exert its effects by promoting a decrease in P2X2 receptor expression and modifications in the density of the NOS-ir,ChAT-ir and CalR-ir myenteric neurons.
基金Supported by So Paulo Research Foundation (FAPESP), Proc 05/04752-0
文摘AIM:To examine whether the ob/ob mouse model of obesity is accompanied by enteric nervous system ab-normalities such as altered motility METHODS:The study examined the distribution of the P2X 2 receptor (P2X 2 R) in myenteric neurons of female ob/ob mice. Specifically, we used immunohistochemistry to analyze the co-expression of the P2X 2 R with neuronal nitric oxide synthase (nNOS), choline acetyltrans-ferase (ChAT), and calretinin (CalR) in neurons of the small intestine myenteric plexus in ob/ob and control female mice In these sections, we used scanning confocal microscopy to analyze the co-localization of these markers as well as the neuronal density (cm 2 ) and area profile (μm2) of P2X 2 R-positive neurons In addition, enteric neurons were labeled using the nicotinamide adenine dinucleotide (NA H) diaphorase method and analyzed with light microscopy as an alternate means by which to analyze neuronal density and areaRESULTS:In the present study, we observed a 29 6% increase in the body weight of the ob/ob animals (OG) compared to the control group (CG) In addition, the average small intestine area was increased by approxi-mately 29 6% in the OG compared to the CG Immu-noreactivity (IR) for the P2X 2 R, nNOS, ChAT and CalR was detectable in the myenteric plexus, as well as in the smooth muscle, in both groups This IR appeared to be mainly cytoplasmic and was also associated with the cell membrane of the myenteric plexus neurons, where it outlined the neuronal cell bodies and their processes P2X 2 R-IR was observed to co-localize 100% with that for nNOS, ChAT and CalR in neurons of both groups In the ob/ob group, however, we observed that the neuronal density (neuron/cm 2 ) of P2X 2 R-IR cells was in-creased by 62% compared to CG, while that of NOS-IR and ChAT-IR neurons was reduced by 49% and 57%, respectively, compared to control mice The neuronal density of CalR-IR neurons was not different between the groups Morphometric studies further demonstrated that the cell body profile area (μm2) of nNOS-IR, ChAT-IR and CalR-IR neurons was increased by 34%, 20% and 55%, respectively, in the OG compared to controls Staining for NA H diaphorase activity is widely used to detect alterations in the enteric nervous system; however, our qualitative examination of NA H-diaphorase positive neurons in the myenteric ganglia revealed an overall similarity between the two groups CONCLUSION:We demonstrate increases in P2X2R expression and alterations in nNOS, ChAT and CalR IR in ileal myenteric neurons of female ob/ob mice compared to wild-type controls.
文摘目的通过研究匹罗卡品致癫痫大鼠海马组织中P2X2受体(嘌呤受体)的表达变化,探讨其在颞叶癫痫发病中的作用机制。方法应用氯化锂-匹罗卡品腹腔注射建立大鼠颞叶癫痫模型,并予以P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)腹腔注射,应用Western blot及Real-time PCR技术检测大鼠海马组织P2X2受体的表达,免疫组化技术检测各组大鼠海马组织中谷氨酸(glutamate,GLU)表达水平。结果 Western blot及Real-time PCR结果显示:慢性自发性癫痫发作组中P2X2表达明显增高(P<0.05),BBG干预组表达减低P<0.05)。免疫组化检测显示:GLU在BBG干预组较慢性自发性发作组中释放减少(P<0.05),且与BBG剂量呈负相关性。结论 P2X2受体通路可能参与颞叶癫痫的发病过程,并有望成为新一代治疗颞叶癫痫的药物靶点。
文摘目的通过研究P2X2受体(嘌呤受体)蛋白在颞叶癫痫大鼠脑组织中的表达水平及星形胶质细胞的增殖水平,以及应用P2X2受体拮抗剂亮蓝G(Brilliant Blue G,BBG)后对颞叶癫痫的影响,探讨P2X2受体在颞叶癫痫发病机制中的作用。方法应用氯化锂-匹罗卡品腹腔注射建立SD大鼠颞叶癫痫模型,将大鼠分为正常盐水对照组、急性发作组、慢性自发性发作组;其中慢性自发性发作组又分为空白对照组(生理盐水)和亮蓝G干预组,应用免疫组化检测各组大鼠海马区P2X2受体及胶质纤维酸性蛋白(GFAP)的表达水平,GFAP反映星形胶质细胞的增殖水平。结果免疫组化检测发现,P2X2受体主要表达于大鼠海马组织的CA1、CA3、齿状回、门区,并且在丘脑也有一定的表达;P2X2受体及GFAP在慢性自发性发作空白对照组中的表达明显高于正常盐水对照组和急性发作组(P<0.05);而亮蓝G干预组与慢性自发性发作空白对照组相比,P2X2受体及GFAP表达水平明显降低(P<0.05)。结论 P2X2受体可能在颞叶癫痫的发病机制中有重要作用,阻滞P2X2受体可能是治疗颞叶癫痫的潜在靶点。
基金supported by Deutsche Forschungsgemeinschaft(DFGIL 20/21-1)Sino-German Centre(GZ919)
文摘Neural stem/progenitor cells：Radial glial cells constitute multipotent cells in the ventricular zone,lining the wall of the lateral ventricle of the embryonic brain.They have the capacity to give rise to cells belonging to all three major linages（neurons,astrocytes and oligodendrocytes）of the nervous system（Tang and Illes,2017）.
基金funded by the National Natural Science Foundation of China,No.30973783the Shanghai Municipal Scientific Committee Project,No.11ZR1434300the Key Subject Program of State Administration of Traditional Chinese Medicine
文摘Electroacupuncture at Shangjuxu （ST37） and Tianshu （ST25） can improve visceral hypersensitivity in rats. Colorectal distension was used to establish a rat model of chronic visceral hypersensitivity. Immunohistochemistry was used to detect P2X2 and P2X3 receptor expression in dorsal root ganglia from rats with chronic visceral hypersensitivity. Results demonstrated that abdominal withdrawal reflex scores obviously increased following establishment of the model, indicating visceral hypersensitivity. Simultaneously, P2X2 and P2X3 receptor expression increased in dorsal root ganglia. After bilateral electroacupuncture at Shangjuxu and Tianshu, abdominal withdrawal reflex scores and P2X2 and P2X3 receptor expression decreased in rats with visceral hypersensitivity. These results indicated that electroacupuncture treatment improved visceral hypersensitivity in rats with irritable bowel syndrome by reducing P2X2 and P2X3 receptor expression in dorsal root ganglia.
基金Project supported by the National Natural Science Foundation of China(Nos.81771208 and 81971043)the Health and Family Planning Commission of Wuxi(No.YGZXM1406)+3 种基金the Wuxi Municipal Bureau on Science and Technology(No.CSE31N1614)the Fundamental Research Fund of Wuxi People’s Hospital(No.RKA201720)the Technology for Social Development Project of Kunshan(No.KS1539)China.
文摘Painful diabetic neuropathy(PDN)is a diabetes mellitus complication.Unfortunately,the mechanisms underlying PDN are still poorly understood.Adenosine triphosphate(ATP)-gated P2X7 receptor(P2X7R)plays a pivotal role in non-diabetic neuropathic pain,but little is known about its effects on streptozotocin(STZ)-induced peripheral neuropathy.Here,we explored whether spinal cord P2X7R was correlated with the generation of mechanical allodynia(MA)in STZ-induced type 1 diabetic neuropathy in mice.MA was assessed by measuring paw withdrawal thresholds and western blotting.Immunohistochemistry was applied to analyze the protein expression levels and localization of P2X7R.STZ-induced mice expressed increased P2X7R in the dorsal horn of the lumbar spinal cord during MA.Mice injected intrathecally with a selective antagonist of P2X7R and P2X7R knockout(KO)mice both presented attenuated progression of MA.Double-immunofluorescent labeling demonstrated that P2X7R-positive cells were mostly co-expressed with Iba1(a microglia marker).Our results suggest that P2X7R plays an important role in the development of MA and could be used as a cellular target for treating PDN.
基金supported by the National Natural Science Foundation of China,No.81260190the Natural Science Foundation of Jiangxi Province of China,No.20132BAB205023+1 种基金a grant from the Science and Technology Research Program of Department of Education of Jiangxi Province in China,No.GJJ13159a grant from the Science and Technology Program of Department of Health of Jiangxi Province,No.20132019
文摘Olfactory bulb tissue transplantation inhibits P2X2/3 receptor-mediated neuropathic pain. However, the olfactory bulb has a complex cellular composition, and the mechanism underlying the action of purified transplanted olfactory ensheathing cells（OECs） remains unclear. In the present study, we microencapsulated OECs in alginic acid, and transplanted free and microencapsulated OECs into the region surrounding the injured sciatic nerve in rat models of chronic constriction injury. We assessed mechanical nociception in the rat models 7 and 14 days after surgery by measuring paw withdrawal threshold, and examined P2X2/3 receptor expression in L4–5 dorsal root ganglia using immunohistochemistry. Rats that received free and microencapsulated OEC transplants showed greater withdrawal thresholds than untreated model rats, and weaker P2X2/3 receptor immunoreactivity in dorsal root ganglia. At 14 days, paw withdrawal threshold was much higher in the microencapsulated OEC-treated animals. Our results confirm that microencapsulated OEC transplantation suppresses P2X2/3 receptor expression in L4–5 dorsal root ganglia in rat models of neuropathic pain and reduces allodynia, and also suggest that transplantation of microencapsulated OECs is more effective than transplantation of free OECs for the treatment of neuropathic pain.
