针刺与运动不同干预次序组合对胆红素脑病大鼠大脑皮层BDNF表达的影响
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摘要
目的:探讨针刺与运动不同干预次序组合治疗胆红素脑病大鼠模型对大鼠大脑皮层BDNF表达的影响,为治疗胆红素脑病提供实验室依据。
方法:选择20只SD大鼠幼鼠,随机分为4组:正常组,模型对照组,先针刺后运动组(AE),先运动后针刺组(EA)。其中正常组以生理盐水行腹腔注射,其余三组均采用胆红素造模法制胆红素脑病模型。模型对照组不进行治疗。治疗组在造模成功后的次日进行治疗:针刺选取双侧顶颞前斜线,运功疗法采用滚筒训练法,每次治疗40分钟,针刺和运动各20分钟,1次/天,疗程1周。疗程结束后,按照原位取材法,取全脑组织,进行组织修块,取中间部分大脑皮层区,经前固定→漂洗→脱水→浸透→包埋、聚合→切片、免疫组化等制样过程,在万能电子显微镜下观察,借助Motic ImagesAdvanced3.0摄片,Motic Images Advanced6.0医学图像分析系统测量免疫组化阳性细胞,并分别统计出各组大脑皮层BDNF的阳性细胞数。比较各组大脑皮层BDNF的阳性细胞数,做统计分析。
结果:(1)正常组幼鼠未出现反应迟钝、俯卧、抽搐等现象;模型组幼鼠均出现反应迟钝、俯卧、抽搐等。大脑皮层镜下观察可见免疫组化染成黄色的BDNF阳性细胞,模型对照组及治疗组可见到胆红素沉积。(2)①模型组(180.40±6.88)较正常组(89.95±6.52)BDNF阳性细胞数有非常显著性差异(P<0.01);②先针刺后运动组(127.81±3.13)与正常组(89.95±6.52)比较有非常显著性差异(P<0.01);③先针刺后运动组(127.81±3.13)与模型组(1.07±0.03)比较有非常显著性差异(P<0.01);④先运动后针刺组(136.59±10.52)与正常组(89.95±6.52)比较有非常显著性差异P<0.01);⑤先运动后针刺组(136.59±10.52)与模型组(180.40±6.88)比较有非常显著性差异(P<0.01)⑥先针刺后运动组(127.81±3.13)与先运动后针刺组(136.59±10.52)比较有正常显著性差异(P<0.01)。
结论:治疗组经过治疗后大鼠反应较前好转,活动灵活,脑源性神经营养因子(BDNF)阳性细胞较模型组降低,但仍高于正常组。针刺及运动治疗能促进幼鼠的恢复,改善脑功能,从而使BDNF的阳性表达下降,提示治疗有效。两组不同的干预次序组合疗效对比有显著性差异(P<0.01),先针刺后运动组BDNF的阳性表达少于先运动后针刺组,提示AE组疗效更优于EA组。
Purpose: To explore the effects of Acupuncture and exercise in the different order of combination treatment interventions rat model of bilirubin encephalopathy in rat cerebral cortex BDNF expression, for the treatment of bilirubin encephalopathy in accordance with the provision of laboratory.
Method: Select 20 young SD rats were randomly divided into 4 groups: normal group, model control group, exercise after the first acupuncture group (AE), acupuncture group after the first exercise (EA). To one of normal saline by intraperitoneal injection line, and the remaining three groups are the rule of law model bilirubin bilirubin encephalopathy model. Model untreated control group. Model in the treatment group the day after the success of treatment: Acupuncture select Top bilateral temporal pre-slash, exercise therapy training method used, each 40-minute treatment, acupuncture and exercise the 20 minutes, 1 second / day , 1 week course of treatment. After the treatment, in accordance with the in situ method based on taking the whole brain tissue, the block repair organizations, take part of the cerebral cortex between areas, by the former fixed dehydration→rinsing→immersion embedding, polymerization→biopsy, Immunohistochemistry, such as sample preparation process at universal electron microscopy, using Motic Images Advanced 3.0 X-ray film, Motic Images Advanced6.0 medical image analysis system for measurement of immunohistochemical-positive cells, and the statistics of each group separately in cerebral cortex of BDNF-positive cells. Comparison of the cerebral cortex in each group the number of BDNF positive cells, to do statistical analysis.
Result: (1) normal rats does not appear insensitive, prone, convulsions and other phenomena; model group rats showed a bit too slow, prone position, such as convulsions. Cerebral cortex were observed dye visible by immunohistochemistry of BDNF positive cells in yellow, model control group and treatment group bilirubin deposition can be seen. (2)①model group (180.40±6.88) than the normal group (89.95±6.52) BDNF-positive cells have a very significant difference (P <0.01);②first acupuncture group after exercise (127.81±3.13) and normal group ( 89.95±6.52) compared have very significant difference (P <0.01);③first acupuncture group after exercise (127.81±3.13) with the model group (1.07±0.03) compared have very significant difference (P <0.01);④first acupuncture group after exercise (136.59±10.52) and normal group (89.95±6.52) more significant difference P <0.01);⑤first acupuncture group after exercise (136.59±10.52) with the model group (180.40±6.88) compared there is a very significant difference (P <0.01)⑥first acupuncture group after exercise (127.81±3.13) after exercise with the first acupuncture group (136.59±10.52) compared there is a very significant difference (P <0.01).
Conclusion: After the treatment group after treatment of rats improved over the previous response, a flexible activity, brain-derived neurotrophic factor (BDNF)-positive cells lower than the model group, but still higher than the normal group. Acupuncture and exercise therapy can promote the recovery of young rats to improve brain function, so that the positive expression of BDNF declined, suggesting that effective treatment. The order of two different combinations of intervention contrast effects were significantly different (P <0.01), exercise group after the first acupuncture positive expression of BDNF is less than the first acupuncture group after exercise, suggesting that more is better than AE Group EA group efficacy.
方法:选择20只SD大鼠幼鼠,随机分为4组:正常组,模型对照组,先针刺后运动组(AE),先运动后针刺组(EA)。其中正常组以生理盐水行腹腔注射,其余三组均采用胆红素造模法制胆红素脑病模型。模型对照组不进行治疗。治疗组在造模成功后的次日进行治疗:针刺选取双侧顶颞前斜线,运功疗法采用滚筒训练法,每次治疗40分钟,针刺和运动各20分钟,1次/天,疗程1周。疗程结束后,按照原位取材法,取全脑组织,进行组织修块,取中间部分大脑皮层区,经前固定→漂洗→脱水→浸透→包埋、聚合→切片、免疫组化等制样过程,在万能电子显微镜下观察,借助Motic ImagesAdvanced3.0摄片,Motic Images Advanced6.0医学图像分析系统测量免疫组化阳性细胞,并分别统计出各组大脑皮层BDNF的阳性细胞数。比较各组大脑皮层BDNF的阳性细胞数,做统计分析。
结果:(1)正常组幼鼠未出现反应迟钝、俯卧、抽搐等现象;模型组幼鼠均出现反应迟钝、俯卧、抽搐等。大脑皮层镜下观察可见免疫组化染成黄色的BDNF阳性细胞,模型对照组及治疗组可见到胆红素沉积。(2)①模型组(180.40±6.88)较正常组(89.95±6.52)BDNF阳性细胞数有非常显著性差异(P<0.01);②先针刺后运动组(127.81±3.13)与正常组(89.95±6.52)比较有非常显著性差异(P<0.01);③先针刺后运动组(127.81±3.13)与模型组(1.07±0.03)比较有非常显著性差异(P<0.01);④先运动后针刺组(136.59±10.52)与正常组(89.95±6.52)比较有非常显著性差异P<0.01);⑤先运动后针刺组(136.59±10.52)与模型组(180.40±6.88)比较有非常显著性差异(P<0.01)⑥先针刺后运动组(127.81±3.13)与先运动后针刺组(136.59±10.52)比较有正常显著性差异(P<0.01)。
结论:治疗组经过治疗后大鼠反应较前好转,活动灵活,脑源性神经营养因子(BDNF)阳性细胞较模型组降低,但仍高于正常组。针刺及运动治疗能促进幼鼠的恢复,改善脑功能,从而使BDNF的阳性表达下降,提示治疗有效。两组不同的干预次序组合疗效对比有显著性差异(P<0.01),先针刺后运动组BDNF的阳性表达少于先运动后针刺组,提示AE组疗效更优于EA组。
Purpose: To explore the effects of Acupuncture and exercise in the different order of combination treatment interventions rat model of bilirubin encephalopathy in rat cerebral cortex BDNF expression, for the treatment of bilirubin encephalopathy in accordance with the provision of laboratory.
Method: Select 20 young SD rats were randomly divided into 4 groups: normal group, model control group, exercise after the first acupuncture group (AE), acupuncture group after the first exercise (EA). To one of normal saline by intraperitoneal injection line, and the remaining three groups are the rule of law model bilirubin bilirubin encephalopathy model. Model untreated control group. Model in the treatment group the day after the success of treatment: Acupuncture select Top bilateral temporal pre-slash, exercise therapy training method used, each 40-minute treatment, acupuncture and exercise the 20 minutes, 1 second / day , 1 week course of treatment. After the treatment, in accordance with the in situ method based on taking the whole brain tissue, the block repair organizations, take part of the cerebral cortex between areas, by the former fixed dehydration→rinsing→immersion embedding, polymerization→biopsy, Immunohistochemistry, such as sample preparation process at universal electron microscopy, using Motic Images Advanced 3.0 X-ray film, Motic Images Advanced6.0 medical image analysis system for measurement of immunohistochemical-positive cells, and the statistics of each group separately in cerebral cortex of BDNF-positive cells. Comparison of the cerebral cortex in each group the number of BDNF positive cells, to do statistical analysis.
Result: (1) normal rats does not appear insensitive, prone, convulsions and other phenomena; model group rats showed a bit too slow, prone position, such as convulsions. Cerebral cortex were observed dye visible by immunohistochemistry of BDNF positive cells in yellow, model control group and treatment group bilirubin deposition can be seen. (2)①model group (180.40±6.88) than the normal group (89.95±6.52) BDNF-positive cells have a very significant difference (P <0.01);②first acupuncture group after exercise (127.81±3.13) and normal group ( 89.95±6.52) compared have very significant difference (P <0.01);③first acupuncture group after exercise (127.81±3.13) with the model group (1.07±0.03) compared have very significant difference (P <0.01);④first acupuncture group after exercise (136.59±10.52) and normal group (89.95±6.52) more significant difference P <0.01);⑤first acupuncture group after exercise (136.59±10.52) with the model group (180.40±6.88) compared there is a very significant difference (P <0.01)⑥first acupuncture group after exercise (127.81±3.13) after exercise with the first acupuncture group (136.59±10.52) compared there is a very significant difference (P <0.01).
Conclusion: After the treatment group after treatment of rats improved over the previous response, a flexible activity, brain-derived neurotrophic factor (BDNF)-positive cells lower than the model group, but still higher than the normal group. Acupuncture and exercise therapy can promote the recovery of young rats to improve brain function, so that the positive expression of BDNF declined, suggesting that effective treatment. The order of two different combinations of intervention contrast effects were significantly different (P <0.01), exercise group after the first acupuncture positive expression of BDNF is less than the first acupuncture group after exercise, suggesting that more is better than AE Group EA group efficacy.
引文
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