P2X4 and P2X7 receptors play an important role in neuropathic pain after spinal cord injury. Regulation of P2X4 and P2X7 receptors can obviously reduce pain hypersensitivity after injury. To investigate the role of ne...P2X4 and P2X7 receptors play an important role in neuropathic pain after spinal cord injury. Regulation of P2X4 and P2X7 receptors can obviously reduce pain hypersensitivity after injury. To investigate the role of neural stem cell transplantation on P2X receptor-mediated neuropathic pain and explore related mechanisms, a rat model of spinal cord injury was prepared using the free-falling heavy body method with spinal cord segment 10 as the center. Neural stem cells were injected into the injured spinal cord segment using a micro-syringe. Expression levels of P2X4 and P2X7 receptors, neurofilament protein, and glial fibrillary acidic protein were determined by immunohistochemistry and western blot assay. In addition, sensory function was quantitatively assessed by current perception threshold. The Basso-Beattie-Bresnahan locomotor rating scale was used to assess neuropathological pain. The results showed that 4 weeks after neural stem cell transplantation, expression of neurofilament protein in the injured segment was markedly increased, while expression of glial fibrillary acidic protein and P2X4 and P2X7 receptors was decreased. At this time point, motor and sensory functions of rats were obviously improved, and neuropathic pain was alleviated. These findings demonstrated that neural stem cell transplantation reduced overexpression of P2X4 and P2X7 receptors, activated locomotor and sensory function reconstruction, and played an important role in neuropathic pain regulation after spinal cord injury. Therefore, neural stem cell transplantation is one potential option for relieving neuropathic pain mediated by P2X receptors.展开更多
目的探讨黄芩苷对糖尿病大鼠视网膜小胶质细胞的激活作用及其机制。方法大鼠腹腔注射链脲佐菌素建立糖尿病模型,随机分为模型组(MG)和黄芩苷组(BG),另将健康大鼠设对照组(CG),每组12只。BG组大鼠给予黄芩苷水溶液(50 mg/kg)每日灌胃,CG...目的探讨黄芩苷对糖尿病大鼠视网膜小胶质细胞的激活作用及其机制。方法大鼠腹腔注射链脲佐菌素建立糖尿病模型,随机分为模型组(MG)和黄芩苷组(BG),另将健康大鼠设对照组(CG),每组12只。BG组大鼠给予黄芩苷水溶液(50 mg/kg)每日灌胃,CG、MG组大鼠给予同体积的蒸馏水灌胃,连续给药12周。12周后测量大鼠血糖,然后麻醉取右眼眼球,苏木精-伊红(HE)染色观察视网膜形态,免疫组织化学法检测视网膜小胶质细胞簇分化抗原68(CD68)蛋白表达,免疫荧光法检测信号转导和转录激活因子3(STAT3)蛋白表达,实时荧光定量PCR(RT-q PCR)检测白细胞介素-6(IL-6)、STAT3 m RNA的相对表达量。结果(1)视网膜形态:与CG组相比,MG组大鼠视网膜神经节细胞(RGC)丢失明显增多,而BG组RGC数量增加;MG组视网膜厚度较CG组薄,而BG组较MG组厚。(2)小胶质细胞:CG组小胶质细胞局限在内核层;MG组小胶质细胞数量增多,且分布在视网膜的每一层中;BG组小胶质细胞数量较MG组少,且小胶质细胞迁移受限。MG组大鼠视网膜小胶质细胞CD68表达高于CG组(t=7.348,P=0.000),BG组CD68表达低于MG组(t=2.449,P=0.019),差异均有统计学意义。(3)STAT3、IL-6:CG组视网膜STAT3表达主要分布在内丛状层,而MG组视网膜各层均有大量STAT3阳性表达,BG组视网膜各层中STAT3表达明显弱于MG组。MG组大鼠视网膜STAT3 m RNA、IL-6 m RNA表达高于CG组(t_(STAT3)=9.935、t_(IL-6)=17.501,均P=0.000),BG组视网膜STAT3 m RNA、IL-6 m RNA表达低于MG组(t_(STAT3)=8.432、t_(IL-6)=9.944,均P=0.000),差异均有统计学意义。结论黄芩苷能抑制糖尿病大鼠视网膜小胶质细胞激活,其机制可能与IL-6/STAT3信号通路有关。展开更多
基金financially supported by the Natural Science Foundation of Shandong Province of China,No.ZR2014HM046(to ZCZ),ZR2015HL113(to XJD),and ZR2014HL101(to XYW)the Science and Technology Development Project of Taian City of China,No.2015NS2183(to XJD)
文摘P2X4 and P2X7 receptors play an important role in neuropathic pain after spinal cord injury. Regulation of P2X4 and P2X7 receptors can obviously reduce pain hypersensitivity after injury. To investigate the role of neural stem cell transplantation on P2X receptor-mediated neuropathic pain and explore related mechanisms, a rat model of spinal cord injury was prepared using the free-falling heavy body method with spinal cord segment 10 as the center. Neural stem cells were injected into the injured spinal cord segment using a micro-syringe. Expression levels of P2X4 and P2X7 receptors, neurofilament protein, and glial fibrillary acidic protein were determined by immunohistochemistry and western blot assay. In addition, sensory function was quantitatively assessed by current perception threshold. The Basso-Beattie-Bresnahan locomotor rating scale was used to assess neuropathological pain. The results showed that 4 weeks after neural stem cell transplantation, expression of neurofilament protein in the injured segment was markedly increased, while expression of glial fibrillary acidic protein and P2X4 and P2X7 receptors was decreased. At this time point, motor and sensory functions of rats were obviously improved, and neuropathic pain was alleviated. These findings demonstrated that neural stem cell transplantation reduced overexpression of P2X4 and P2X7 receptors, activated locomotor and sensory function reconstruction, and played an important role in neuropathic pain regulation after spinal cord injury. Therefore, neural stem cell transplantation is one potential option for relieving neuropathic pain mediated by P2X receptors.
文摘目的探讨黄芩苷对糖尿病大鼠视网膜小胶质细胞的激活作用及其机制。方法大鼠腹腔注射链脲佐菌素建立糖尿病模型,随机分为模型组(MG)和黄芩苷组(BG),另将健康大鼠设对照组(CG),每组12只。BG组大鼠给予黄芩苷水溶液(50 mg/kg)每日灌胃,CG、MG组大鼠给予同体积的蒸馏水灌胃,连续给药12周。12周后测量大鼠血糖,然后麻醉取右眼眼球,苏木精-伊红(HE)染色观察视网膜形态,免疫组织化学法检测视网膜小胶质细胞簇分化抗原68(CD68)蛋白表达,免疫荧光法检测信号转导和转录激活因子3(STAT3)蛋白表达,实时荧光定量PCR(RT-q PCR)检测白细胞介素-6(IL-6)、STAT3 m RNA的相对表达量。结果(1)视网膜形态:与CG组相比,MG组大鼠视网膜神经节细胞(RGC)丢失明显增多,而BG组RGC数量增加;MG组视网膜厚度较CG组薄,而BG组较MG组厚。(2)小胶质细胞:CG组小胶质细胞局限在内核层;MG组小胶质细胞数量增多,且分布在视网膜的每一层中;BG组小胶质细胞数量较MG组少,且小胶质细胞迁移受限。MG组大鼠视网膜小胶质细胞CD68表达高于CG组(t=7.348,P=0.000),BG组CD68表达低于MG组(t=2.449,P=0.019),差异均有统计学意义。(3)STAT3、IL-6:CG组视网膜STAT3表达主要分布在内丛状层,而MG组视网膜各层均有大量STAT3阳性表达,BG组视网膜各层中STAT3表达明显弱于MG组。MG组大鼠视网膜STAT3 m RNA、IL-6 m RNA表达高于CG组(t_(STAT3)=9.935、t_(IL-6)=17.501,均P=0.000),BG组视网膜STAT3 m RNA、IL-6 m RNA表达低于MG组(t_(STAT3)=8.432、t_(IL-6)=9.944,均P=0.000),差异均有统计学意义。结论黄芩苷能抑制糖尿病大鼠视网膜小胶质细胞激活,其机制可能与IL-6/STAT3信号通路有关。
