Inflammation plays an important role in nerve defects caused by intracerebral hemorrhage. Repairing brain damage by inhibiting the macrophage-inducible C-type lectin/spleen tyrosine kinase （Mincle/Syk） signaling pat...Inflammation plays an important role in nerve defects caused by intracerebral hemorrhage. Repairing brain damage by inhibiting the macrophage-inducible C-type lectin/spleen tyrosine kinase （Mincle/Syk） signaling pathway is a potential new target for treating cerebral hemorrhage. In this study, we aimed to determine whether acupuncture through Baihui （DU20） to Qubin （GBT） is an effective treatment for intracerebral hemorrhage through the Mincle/Syk signaling pathway. An intracerebral hemorrhage rat model was established by autol- ogous blood infusion into the caudate nucleus. Acupuncture through Baihui to Qubin was performed for 30 minutes, once every 12 hours, for a total of three times. Piceatannol （34.62 mg/kg）, a Syk inhibitor, was intraperitoneally injected as a control. Modified neurological severity score was used to assess neurological function. Brain water content was measured. Immunohistochemistry and western blot assay were used to detect immunoreactivity and protein expression levels of Minde, Syk, and CARD9. Real-time polymerase chain reaction was used to determine interleukin-1[~ mRNA levels. Hematoxylin-eosin staining was performed to observe histopathological changes. Our re- suits showed that acupuncture through Baihui to Qubin remarkably improved neurological function and brain water content, and inhibited immunoreactivity and expression of Mincle, Syk, CARDg, and interkeukin-1β Moreover, this effect was similar to piceatannol. These find- ings suggest that acupuncture through Baihui to Qubin can improve neurological impairment after cerebral hemorrhage by inhibiting the Mincle/Syk signaling pathway.展开更多
目的 观察针刺百会、曲鬓穴对急性期脑出血(ICH)大鼠白细胞分化抗原36(CD36)、Toll样受体4(TLR4)表达的影响,探讨针刺治疗脑出血的作用机制。方法 选择144只Wistar雄性大鼠,采用随机数字表法分为假手术组、模型组、针刺组、抑制剂组4组...目的 观察针刺百会、曲鬓穴对急性期脑出血(ICH)大鼠白细胞分化抗原36(CD36)、Toll样受体4(TLR4)表达的影响,探讨针刺治疗脑出血的作用机制。方法 选择144只Wistar雄性大鼠,采用随机数字表法分为假手术组、模型组、针刺组、抑制剂组4组,每组36只,每组按1、3、7 d时间点再分为3个亚组,每组12只。采用立体定位自体血注入法建立ICH大鼠模型。模型组仅接受ICH模型制备,不进行任何治疗;假手术组接受类似模型组各项手术操作,但不进行注血制作;抑制剂组造模后6 h,腹腔注射TLR4抑制剂TAK242,3 mg/kg,1次/d,连续5 d;造模12 h后,针刺组各亚组开始接受针刺治疗,穴位选择百会穴(顶骨正中)、右侧曲鬓穴,采用透刺方法,进针深度20 mm,以100 r/min小幅度捻转,持续捻转2 min,每间隔5 min捻针1次,共留针30 min,期间捻转3次,1次/d,针刺组各亚组分别治疗1、3、7 d。分别于治疗后第1、3、7天,采用改良神经功能缺损评分(mNSS)评估大鼠神经功能;检测脑组织血肿体积;采用Western blot法检测脑组织CD36、TLR4蛋白表达水平;采用免疫荧光法观察CD36、TLR4在星形胶质细胞中的表达。结果 (1) mNSS评分:与假手术组同一时间点比较,模型组、针刺组、抑制剂组治疗后1、3、7 d mNSS评分均明显升高(P<0.05);与模型组同一时间点比较,针刺组、抑制剂组治疗后1、3、7 d mNSS评分均明显降低(P<0.05);与针刺组同一时间点比较,抑制剂组治疗后1 d mNSS评分明显降低(P<0.05)。(2)血肿体积:与模型组同一时间点比较,针刺组、抑制剂组治疗后3、7 d脑血肿体积均明显降低,抑制剂组治疗后1、3、7 d脑血肿体积明显降低(P<0.05);与针刺组同一时间点比较,抑制剂组治疗后1、3、7 d脑血肿体积明显降低(P<0.05)。(3) CD36、TLR4蛋白表达水平:与假手术组同一时间点比较,模型组、针刺组、抑制剂组治疗后1、3、7 d CD36、TLR4蛋白表达水平均明显升高(P<0.05);与模型组同一时间点比较,针刺组、抑制剂组治疗后1、3、7 d CD36蛋白表达水平均明显升高(P<0.05),针刺组、抑制剂组治疗后3、7 d TLR4蛋白表达水平均明显降低(P<0.05);与针刺组比较,抑制剂组治疗后1、3、7 d CD36蛋白表达水平均明显升高,TLR4蛋白表达水平均明显降低(P<0.05)。(4) CD36、TLR4在GFAP中的表达水平:假手术组大鼠脑组织内可见少量CD36、TLR4表达。与假手术组同一时间点比较,模型组治疗后1、3、7 d CD36、TLR4在GFAP表达均明显增加(P<0.05);与模型组同一时间点比较,抑制剂组、针刺组治疗后1、3、7 d CD36在GFAP表达明显增加,TLR4在GFAP表达明显降低(P<0.05)。结论 针刺百会、曲鬓穴可以改善急性期ICH大鼠神经功能,减轻脑出血血肿体积,可能与促进CD36蛋白表达、抑制TLR4蛋白表达有关。展开更多
目的:观察脑出血大鼠血肿周围脑组织神经元特异性烯醇化酶(neuron specific enolase,NSE)的表达,探讨"百会"透"曲鬓"头针疗法治疗脑出血的可能作用机制。方法:选取220只健康雄性W istar大鼠,将其中200只大鼠随机分...目的:观察脑出血大鼠血肿周围脑组织神经元特异性烯醇化酶(neuron specific enolase,NSE)的表达,探讨"百会"透"曲鬓"头针疗法治疗脑出血的可能作用机制。方法:选取220只健康雄性W istar大鼠,将其中200只大鼠随机分为两组:模型组和针刺组,每组100只,每组再随机分为6h、1d、2d、3d、7d五个亚组,每个亚组20只;而剩余20只作为空白组。采用自体血注入法制备脑出血模型;针刺组针刺病灶侧"百会"透"曲鬓"穴。应用神经行为学指标观察各组大鼠神经功能缺损程度;在光镜下观察各组大鼠脑组织病理形态学改变;运用免疫组化方法测定不同时间点各组大鼠脑组织中NSE的表达。结果:①空白组的大鼠表现正常;而各造模组大鼠术后均出现不同程度的神经功能缺损,以3d时最为严重。针刺组大鼠神经功能缺损征较模型组有明显恢复,与模型组比较,2d、3d,7d评分,差异统计学意义(P<0.01)。②光镜下可见模型组血肿周围脑组织坏死、水肿、炎性细胞浸润,神经细胞核固缩、空泡化,毛细血管扩张、充血,3天时最严重。针刺组脑组织逐渐修复,水肿程度较轻,炎性细胞减少。③空白组大鼠脑组织未见NSE阳性细胞表达;而模型组和针刺组大鼠术后均出现不同程度的NSE阳性细胞表达增多现象。模型组大鼠术后6h即出现大量的NSE阳性细胞表达;至3d时达高峰,7d时有所下降。针刺组NSE阳性细胞表达低于模型组,与模型组在相同时间点比较均差异明显(P<0.01)。结论:①脑出血后大鼠脑组织内NSE表达上调,且其表达水平的变化与脑组织病理形态的改变、神经功能缺损程度相关;②"百会"透"曲鬓"针刺法可能是通过降低NSE在脑出血大鼠急性期脑组织中的表达、抑制脑水肿的形成,从而保护受损的神经元细胞、促进缺血半暗带的神经元细胞的存活,以达到脑保护的作用。展开更多
Objective:To explore the effect of acupuncture on the expression of heme oxygenase-1 in rats with acute cerebral hemorrhage.Methods:108 Wistar male rats were randomly divided into sham operation group,model group,and ...Objective:To explore the effect of acupuncture on the expression of heme oxygenase-1 in rats with acute cerebral hemorrhage.Methods:108 Wistar male rats were randomly divided into sham operation group,model group,and acupuncture combined with model group(referred to as acupuncture group).Each group was divided into 3 subgroups according to 1d,3d and 7d,with 6 rats in each subgroup.The rat model of cerebral hemorrhage was established by autologous blood injection.Acupuncture was given at Baihui(GU20)and Qubin(GB7).Separately,at the 1st,3rd and 7th day,modified neurological severity score was used to evaluate the neurological function of rats,HE staining was used to measure the volume of cerebral hematoma and western blot was used to detect the expression of heme oxygenase-1 protein in cerebral hematoma tissue.Results:Compared with the model group,at each time point,the modified neurological severity score of the acupuncture group was significantly reduced(P<0.01);at the two time points of 3rd and 7th day,the cerebral hematoma volume of the acupuncture group was significantly reduced(P<0.05)and the expression of heme oxygenase-1 protein in brain tissue was significantly increased(P<0.05).Conclusion:Acupuncture may promote the expression of heme oxygenase-1 protein,reduce the volume of hematoma and the score of neurological deficit in rats with intracerebral hemorrhage,improve the performance of neurological deficit,and play a role in brain protection.展开更多
基金supported by the National Natural Science Foundation of China,No.81473764,81273824the Key Project of Natural Science Foundation of Heilongjiang Province of China,No.ZD201204the Doctoral Fund Program of Ministry of Education of China,No.20102327110003
文摘Inflammation plays an important role in nerve defects caused by intracerebral hemorrhage. Repairing brain damage by inhibiting the macrophage-inducible C-type lectin/spleen tyrosine kinase （Mincle/Syk） signaling pathway is a potential new target for treating cerebral hemorrhage. In this study, we aimed to determine whether acupuncture through Baihui （DU20） to Qubin （GBT） is an effective treatment for intracerebral hemorrhage through the Mincle/Syk signaling pathway. An intracerebral hemorrhage rat model was established by autol- ogous blood infusion into the caudate nucleus. Acupuncture through Baihui to Qubin was performed for 30 minutes, once every 12 hours, for a total of three times. Piceatannol （34.62 mg/kg）, a Syk inhibitor, was intraperitoneally injected as a control. Modified neurological severity score was used to assess neurological function. Brain water content was measured. Immunohistochemistry and western blot assay were used to detect immunoreactivity and protein expression levels of Minde, Syk, and CARD9. Real-time polymerase chain reaction was used to determine interleukin-1[~ mRNA levels. Hematoxylin-eosin staining was performed to observe histopathological changes. Our re- suits showed that acupuncture through Baihui to Qubin remarkably improved neurological function and brain water content, and inhibited immunoreactivity and expression of Mincle, Syk, CARDg, and interkeukin-1β Moreover, this effect was similar to piceatannol. These find- ings suggest that acupuncture through Baihui to Qubin can improve neurological impairment after cerebral hemorrhage by inhibiting the Mincle/Syk signaling pathway.
文摘目的 观察针刺百会、曲鬓穴对急性期脑出血(ICH)大鼠白细胞分化抗原36(CD36)、Toll样受体4(TLR4)表达的影响,探讨针刺治疗脑出血的作用机制。方法 选择144只Wistar雄性大鼠,采用随机数字表法分为假手术组、模型组、针刺组、抑制剂组4组,每组36只,每组按1、3、7 d时间点再分为3个亚组,每组12只。采用立体定位自体血注入法建立ICH大鼠模型。模型组仅接受ICH模型制备,不进行任何治疗;假手术组接受类似模型组各项手术操作,但不进行注血制作;抑制剂组造模后6 h,腹腔注射TLR4抑制剂TAK242,3 mg/kg,1次/d,连续5 d;造模12 h后,针刺组各亚组开始接受针刺治疗,穴位选择百会穴(顶骨正中)、右侧曲鬓穴,采用透刺方法,进针深度20 mm,以100 r/min小幅度捻转,持续捻转2 min,每间隔5 min捻针1次,共留针30 min,期间捻转3次,1次/d,针刺组各亚组分别治疗1、3、7 d。分别于治疗后第1、3、7天,采用改良神经功能缺损评分(mNSS)评估大鼠神经功能;检测脑组织血肿体积;采用Western blot法检测脑组织CD36、TLR4蛋白表达水平;采用免疫荧光法观察CD36、TLR4在星形胶质细胞中的表达。结果 (1) mNSS评分:与假手术组同一时间点比较,模型组、针刺组、抑制剂组治疗后1、3、7 d mNSS评分均明显升高(P<0.05);与模型组同一时间点比较,针刺组、抑制剂组治疗后1、3、7 d mNSS评分均明显降低(P<0.05);与针刺组同一时间点比较,抑制剂组治疗后1 d mNSS评分明显降低(P<0.05)。(2)血肿体积:与模型组同一时间点比较,针刺组、抑制剂组治疗后3、7 d脑血肿体积均明显降低,抑制剂组治疗后1、3、7 d脑血肿体积明显降低(P<0.05);与针刺组同一时间点比较,抑制剂组治疗后1、3、7 d脑血肿体积明显降低(P<0.05)。(3) CD36、TLR4蛋白表达水平:与假手术组同一时间点比较,模型组、针刺组、抑制剂组治疗后1、3、7 d CD36、TLR4蛋白表达水平均明显升高(P<0.05);与模型组同一时间点比较,针刺组、抑制剂组治疗后1、3、7 d CD36蛋白表达水平均明显升高(P<0.05),针刺组、抑制剂组治疗后3、7 d TLR4蛋白表达水平均明显降低(P<0.05);与针刺组比较,抑制剂组治疗后1、3、7 d CD36蛋白表达水平均明显升高,TLR4蛋白表达水平均明显降低(P<0.05)。(4) CD36、TLR4在GFAP中的表达水平:假手术组大鼠脑组织内可见少量CD36、TLR4表达。与假手术组同一时间点比较,模型组治疗后1、3、7 d CD36、TLR4在GFAP表达均明显增加(P<0.05);与模型组同一时间点比较,抑制剂组、针刺组治疗后1、3、7 d CD36在GFAP表达明显增加,TLR4在GFAP表达明显降低(P<0.05)。结论 针刺百会、曲鬓穴可以改善急性期ICH大鼠神经功能,减轻脑出血血肿体积,可能与促进CD36蛋白表达、抑制TLR4蛋白表达有关。
文摘目的:观察脑出血大鼠血肿周围脑组织神经元特异性烯醇化酶(neuron specific enolase,NSE)的表达,探讨"百会"透"曲鬓"头针疗法治疗脑出血的可能作用机制。方法:选取220只健康雄性W istar大鼠,将其中200只大鼠随机分为两组:模型组和针刺组,每组100只,每组再随机分为6h、1d、2d、3d、7d五个亚组,每个亚组20只;而剩余20只作为空白组。采用自体血注入法制备脑出血模型;针刺组针刺病灶侧"百会"透"曲鬓"穴。应用神经行为学指标观察各组大鼠神经功能缺损程度;在光镜下观察各组大鼠脑组织病理形态学改变;运用免疫组化方法测定不同时间点各组大鼠脑组织中NSE的表达。结果:①空白组的大鼠表现正常;而各造模组大鼠术后均出现不同程度的神经功能缺损,以3d时最为严重。针刺组大鼠神经功能缺损征较模型组有明显恢复,与模型组比较,2d、3d,7d评分,差异统计学意义(P<0.01)。②光镜下可见模型组血肿周围脑组织坏死、水肿、炎性细胞浸润,神经细胞核固缩、空泡化,毛细血管扩张、充血,3天时最严重。针刺组脑组织逐渐修复,水肿程度较轻,炎性细胞减少。③空白组大鼠脑组织未见NSE阳性细胞表达;而模型组和针刺组大鼠术后均出现不同程度的NSE阳性细胞表达增多现象。模型组大鼠术后6h即出现大量的NSE阳性细胞表达;至3d时达高峰,7d时有所下降。针刺组NSE阳性细胞表达低于模型组,与模型组在相同时间点比较均差异明显(P<0.01)。结论:①脑出血后大鼠脑组织内NSE表达上调,且其表达水平的变化与脑组织病理形态的改变、神经功能缺损程度相关;②"百会"透"曲鬓"针刺法可能是通过降低NSE在脑出血大鼠急性期脑组织中的表达、抑制脑水肿的形成,从而保护受损的神经元细胞、促进缺血半暗带的神经元细胞的存活,以达到脑保护的作用。
文摘Objective:To explore the effect of acupuncture on the expression of heme oxygenase-1 in rats with acute cerebral hemorrhage.Methods:108 Wistar male rats were randomly divided into sham operation group,model group,and acupuncture combined with model group(referred to as acupuncture group).Each group was divided into 3 subgroups according to 1d,3d and 7d,with 6 rats in each subgroup.The rat model of cerebral hemorrhage was established by autologous blood injection.Acupuncture was given at Baihui(GU20)and Qubin(GB7).Separately,at the 1st,3rd and 7th day,modified neurological severity score was used to evaluate the neurological function of rats,HE staining was used to measure the volume of cerebral hematoma and western blot was used to detect the expression of heme oxygenase-1 protein in cerebral hematoma tissue.Results:Compared with the model group,at each time point,the modified neurological severity score of the acupuncture group was significantly reduced(P<0.01);at the two time points of 3rd and 7th day,the cerebral hematoma volume of the acupuncture group was significantly reduced(P<0.05)and the expression of heme oxygenase-1 protein in brain tissue was significantly increased(P<0.05).Conclusion:Acupuncture may promote the expression of heme oxygenase-1 protein,reduce the volume of hematoma and the score of neurological deficit in rats with intracerebral hemorrhage,improve the performance of neurological deficit,and play a role in brain protection.
