目的探讨电针足三里穴促胃动力作用机制。方法采用电生理学方法同步观察电针不同穴位后胃电的变化;采用免疫组化荧光双重标记胃电起搏区Cajal间质细胞(interstitial cells of cajal,ICCs)与缝隙连接蛋白43(CX43)、ICCs与ERK的方法 ,观...目的探讨电针足三里穴促胃动力作用机制。方法采用电生理学方法同步观察电针不同穴位后胃电的变化;采用免疫组化荧光双重标记胃电起搏区Cajal间质细胞(interstitial cells of cajal,ICCs)与缝隙连接蛋白43(CX43)、ICCs与ERK的方法 ,观察针刺足三里穴引起胃电起搏区ICCs的变化、与ICCs信息传递相关的CX43的变化、丝裂原活化蛋白激酶(MAPK)家族中的ERK的变化情况。结果电针足三里穴对胃电有明显影响。电针足三里穴可使胃电频率及波幅增高;电针足三里穴可显著激活胃ICCs表达及与ICCs信息传递密切相关的CX43的表达;电针足三里穴能促进胃ICCs调节胃运动的细胞信号转导的MAPK途径中ERK的表达。结论电针足三里穴对大鼠胃电具有明显的促进作用;电针足三里穴促进胃运动的机制可能通过激活ICCs而产生显著电生理活动,通过ICCs及SMC之间的缝隙连接蛋白传递达到平滑肌进而调节胃运动。这一作用的完成,可能是通过ICCs信号转导MAPK途径中ERK通路实现的。展开更多
Scutellarin, a bioactive flavone isolated from Scutellaria baicalensis, has anti-inflammatory, anti-neurotoxic, anti-apoptotic and anti-oxida- tive effects and has been used to treat cardiovascular and cerebrovascular...Scutellarin, a bioactive flavone isolated from Scutellaria baicalensis, has anti-inflammatory, anti-neurotoxic, anti-apoptotic and anti-oxida- tive effects and has been used to treat cardiovascular and cerebrovascular diseases in China. However, the mechanisms by which scutellarin mediates neuroprotection in cerebral ischemia remain unclear. The interaction between scutellarin and nicotinamide adenine dinucleotide phosphate oxidase 2 （NOX2） was assessed by molecular docking study, which showed that scutellarin selectively binds to NOX2 with high affinity. Cultures of primary astrocytes isolated from the cerebral cortex of neonatal Sprague-Dawley rats were pretreated with 2, 10 or 50 μM scutellarin for 30 minutes. The astrocytes were then subjected to oxygen/glucose deprivation by incubation for 2 hours in glucose-free Dulbecco＇s modified Eagle＇s medium in a 95% N2/5% CO2 incubator, followed by simulated reperfusion for 22 hours. Cell viability was assessed by cell counting kit-8 assay. Expression levels of NOX2, connexin 43 and caspase-3 were assessed by western blot assay. Reactive oxygen species were measured spectrophotometrically. Pretreatment with 10 or 50 μM scutellarin substantially increased viability, reduced the expression of NOX2 and caspase-3, increased the expression of connexin 43, and diminished the levels of reactive oxygen, species in astrocytes subjected to ischemia-＇reperfusion. We also assessed the effects of scutellarin in vivo in the rat transient middle cerebral artery occlusion model of cerebral ischemia-reperfusion injury. Rats were given intraperitoneal injection of 100 mg/kg scutellarin 2 hours before surgery. The Bederson scale was used to assess neurological deficit, and 2,3,5-triphenyltetrazolium chloride staining was used to measure infarct size. Western blot assay was used to assess expression of NOX2 and connexin 43 in brain tissue. Enzyme-linked immunosorbent assay was used to detect 8-hydroxydeoxyguanosine （8-OHdG）, 4-hydroxy-2-nonenal （4-HNE） and 3-nitrotyr展开更多
文摘目的探讨电针足三里穴促胃动力作用机制。方法采用电生理学方法同步观察电针不同穴位后胃电的变化;采用免疫组化荧光双重标记胃电起搏区Cajal间质细胞(interstitial cells of cajal,ICCs)与缝隙连接蛋白43(CX43)、ICCs与ERK的方法 ,观察针刺足三里穴引起胃电起搏区ICCs的变化、与ICCs信息传递相关的CX43的变化、丝裂原活化蛋白激酶(MAPK)家族中的ERK的变化情况。结果电针足三里穴对胃电有明显影响。电针足三里穴可使胃电频率及波幅增高;电针足三里穴可显著激活胃ICCs表达及与ICCs信息传递密切相关的CX43的表达;电针足三里穴能促进胃ICCs调节胃运动的细胞信号转导的MAPK途径中ERK的表达。结论电针足三里穴对大鼠胃电具有明显的促进作用;电针足三里穴促进胃运动的机制可能通过激活ICCs而产生显著电生理活动,通过ICCs及SMC之间的缝隙连接蛋白传递达到平滑肌进而调节胃运动。这一作用的完成,可能是通过ICCs信号转导MAPK途径中ERK通路实现的。
基金financially supported by the National Natural Science Foundation of China,No.81303115,81774042,81771353the Natural Science Foundation of Guangdong Province of China,No.S2013040016915+3 种基金the Science and Technology Program of Guangzhou City of China,No.201508020050,201604020003the Pearl River S&T Nova Program of Guangzhou,the Postdoctoral Foundation of China,No.BBK42913K09,201003345,BBH429151701a grant from the Hong Kong Scholar Program,Guangzhou University of TCM 2017 High Level University Construction Program,No.A1-AFD018171Z11096a grant from the Specialty Program of Guangdong Province Hospital of Traditional Chinese Medicine of China,No.YN2016MJ07,YN2015QN16,YN2015B2025
文摘Scutellarin, a bioactive flavone isolated from Scutellaria baicalensis, has anti-inflammatory, anti-neurotoxic, anti-apoptotic and anti-oxida- tive effects and has been used to treat cardiovascular and cerebrovascular diseases in China. However, the mechanisms by which scutellarin mediates neuroprotection in cerebral ischemia remain unclear. The interaction between scutellarin and nicotinamide adenine dinucleotide phosphate oxidase 2 （NOX2） was assessed by molecular docking study, which showed that scutellarin selectively binds to NOX2 with high affinity. Cultures of primary astrocytes isolated from the cerebral cortex of neonatal Sprague-Dawley rats were pretreated with 2, 10 or 50 μM scutellarin for 30 minutes. The astrocytes were then subjected to oxygen/glucose deprivation by incubation for 2 hours in glucose-free Dulbecco＇s modified Eagle＇s medium in a 95% N2/5% CO2 incubator, followed by simulated reperfusion for 22 hours. Cell viability was assessed by cell counting kit-8 assay. Expression levels of NOX2, connexin 43 and caspase-3 were assessed by western blot assay. Reactive oxygen species were measured spectrophotometrically. Pretreatment with 10 or 50 μM scutellarin substantially increased viability, reduced the expression of NOX2 and caspase-3, increased the expression of connexin 43, and diminished the levels of reactive oxygen, species in astrocytes subjected to ischemia-＇reperfusion. We also assessed the effects of scutellarin in vivo in the rat transient middle cerebral artery occlusion model of cerebral ischemia-reperfusion injury. Rats were given intraperitoneal injection of 100 mg/kg scutellarin 2 hours before surgery. The Bederson scale was used to assess neurological deficit, and 2,3,5-triphenyltetrazolium chloride staining was used to measure infarct size. Western blot assay was used to assess expression of NOX2 and connexin 43 in brain tissue. Enzyme-linked immunosorbent assay was used to detect 8-hydroxydeoxyguanosine （8-OHdG）, 4-hydroxy-2-nonenal （4-HNE） and 3-nitrotyr
