针刺对缺血性脑损伤模型大鼠TNF-α、Caspase-3表达的影响
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摘要
目的：观察针刺督脉大椎、百会、人中(穴)对大鼠缺血区脑组织抗损伤与保护作用。观察缺血时脑组织TNF-α和Caspase-3表达的影响,探讨针剌督脉大椎、百会、人中(穴)对脑组织产生的保护作用及产生这种保护作用的可能机制。
方法：SD雄性大鼠32只随机分为模型组、假手术组、对照点组和针刺组四组,每组各8只。建立MCAO大鼠模型,等MCAO大鼠模型生命体征稳定后,采用ZeaLonga评分法神经功能缺损评分以验证模型成功与否,造模成功者六次治疗后再行神经功能缺损评分,采用HE染色观察大鼠脑组织的病理变化、原味末端标记法(TUNEL)法检测神经细胞凋亡,并应用SP免疫组织化学方法检测大鼠脑缺血区大脑组织中的TNF-α和caspase-3的表达。
结果：1.神经功能评分：①与假手术组比较,模型组、对照点组、穴位组神经功能缺损评分明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,对照组及穴位组神经功能缺损评分降低,穴位组差异有非常显著性意义(P<0.01),对照点组比较差异有显著性意义(P<0.05)；③与对照点组比较,穴位组神经功能缺损评分明显降低,差异有显著性意义(P<0.05)
2.①与假手术组比较,模型组、对照点组、穴位组TNF-α阳性细胞数明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,对照点组穴位组TNF-α阳性细胞数降低,差异有显著性意义(P<0.05),穴位组TNF-α阳性细胞数降低,差异有非常显著性意义(P<0.01)；③与对照点组比较,穴位组TNF-α阳性细胞数降低,差异有显著性意义(P<0.05)
3.①与假手术组比较,模型组、对照点组、穴位组Caspase-3阳性细胞数明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,穴位组的Caspase-3阳性细胞数降低,差异有非常显著性意义(P<0.01),对照点组的Caspase-3阳性细胞数降低不明显,差异无意义(P>0.05)；③与对照点组比较,穴位组Caspase-3阳性细胞数明显降低,差异有非常显著性意义(P<0.01)
结论：1.脑缺血损伤引起大鼠明显的神经功能缺损体征,缺血区神经细胞存在细胞凋亡,而针刺督脉“人中”,“百会”,“大椎”穴可以明显改善大鼠神经功能,有效抑制缺血后脑神经细胞凋亡的发生。
2.针刺督脉“人中”,“百会”,“大椎”穴可以通过抑制神经细胞的炎症因子TNF-α以及级联反应执行凋亡蛋白酶Caspase-3,从而延缓了缺血损伤神经元的死亡,发挥对缺血性脑损伤的保护作用。
Objective:
Observe the effect of acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui of Resistance to injury and protection of the ischemic area. Observed ischemic brain tissue TNF-α and Caspase-3expression to explore acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui on the brain to produce the protective effect and possible mechanisms for this protective effect。
Methods:
SD male rats32were randomly divided into model group, sham operation group, DuiZhaoDian group and acupuncture group of four groups and each group of each only8.The establishment of a MCAO rat model, MCAO rat model of vital signs were stable after ZeaLonga score neurological deficit scores to verify the success of the model or not, six treatment of neurological function after the line with HE staining in rat brain tissue pathological changes in the flavor end labeling (TUNEL) assay detection of neuronal apoptosis, and apply the SP immunohistochemistry was detected in rat focal cerebral ischemia in brain tissue TNF-alpha and caspase-3positive cells.
Result:
1.Nerve functional score:(1) group compared with sham operation group, model group, DuiZhaoDian group, acupuncture group of neural function defect scale increased significantly, difference is very significant meaning (P<0.01);(2)group compared with model, DuiZhaoDian group, acupuncture group of neural function defect score lower, and the meridian group significantly different meaning (P<0.01);DuiZhaoDian group is significant difference(P<0.05);(3)group compared with DuiZhaoDian, acupuncture group of neural function defect scale reduced significantly, and significant difference(P<0.05).
2.Epidemic photochemical staining study dyeing:TNF alpha immune response results show that the number of positive cells group compared with sham operation group, model group DuiZhaoDian group of TNF alpha positive points cell count was higher, meaning with significant difference (P<0.01); Group compared with model, DuiZhaoDian group of TNF alpha positive points lower cell count, the difference was statistically significant (P<0.05), and the meridian group of TNF alpha positive cell count, reduce the significant difference (P<0.01); And DuiZhaoDian group compared, and the meridian group of TNF alpha positive cell count, reduce, the difference was statistically significant (P<0.05)
3.Image analysis ischemia side caspase-3immune response brain cell count and positive results:group compared with sham operation group, model group DuiZhaoDian group caspase-3points group was higher number of positive cells, meaning with significant difference (P<0.01); Group compared with model, and the meridian of the group Caspase-3positive cell count, reduce, the difference was statistically significant (P<0.01), DuiZhaoDian group of Caspase-3positive cells reduce the number is not obvious, meaningless differences (P>0.05); And DuiZhaoDian group compared, and the meridian group Caspase-three positive cell count, reduced significantly, and there is a significant difference of statistical significance (P<0.01).
Conclusion:
1. Ischemic damage rats obvious neural function defect signs, ischemia area nerve cells exist cell apoptosis, and acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui acupuncture point can obviously improve the rat nerve function and effectively restrain brain ischemia cell apoptosis happened after.
2. Acupuncture du meridian people, the Bai Hui, the Ren Zhong, the Da Zhui acupuncture point by inhibiting the neural cells inflammation factors TNF alpha and cascade reaction of the apoptotic protease expression of Caspase-3, delaying the ischemic injury the death of neurons, play to ischemic brain damage of protection.
方法：SD雄性大鼠32只随机分为模型组、假手术组、对照点组和针刺组四组,每组各8只。建立MCAO大鼠模型,等MCAO大鼠模型生命体征稳定后,采用ZeaLonga评分法神经功能缺损评分以验证模型成功与否,造模成功者六次治疗后再行神经功能缺损评分,采用HE染色观察大鼠脑组织的病理变化、原味末端标记法(TUNEL)法检测神经细胞凋亡,并应用SP免疫组织化学方法检测大鼠脑缺血区大脑组织中的TNF-α和caspase-3的表达。
结果：1.神经功能评分：①与假手术组比较,模型组、对照点组、穴位组神经功能缺损评分明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,对照组及穴位组神经功能缺损评分降低,穴位组差异有非常显著性意义(P<0.01),对照点组比较差异有显著性意义(P<0.05)；③与对照点组比较,穴位组神经功能缺损评分明显降低,差异有显著性意义(P<0.05)
2.①与假手术组比较,模型组、对照点组、穴位组TNF-α阳性细胞数明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,对照点组穴位组TNF-α阳性细胞数降低,差异有显著性意义(P<0.05),穴位组TNF-α阳性细胞数降低,差异有非常显著性意义(P<0.01)；③与对照点组比较,穴位组TNF-α阳性细胞数降低,差异有显著性意义(P<0.05)
3.①与假手术组比较,模型组、对照点组、穴位组Caspase-3阳性细胞数明显增高,差异具有非常显著性意义(P<0.01)；②与模型组比较,穴位组的Caspase-3阳性细胞数降低,差异有非常显著性意义(P<0.01),对照点组的Caspase-3阳性细胞数降低不明显,差异无意义(P>0.05)；③与对照点组比较,穴位组Caspase-3阳性细胞数明显降低,差异有非常显著性意义(P<0.01)
结论：1.脑缺血损伤引起大鼠明显的神经功能缺损体征,缺血区神经细胞存在细胞凋亡,而针刺督脉“人中”,“百会”,“大椎”穴可以明显改善大鼠神经功能,有效抑制缺血后脑神经细胞凋亡的发生。
2.针刺督脉“人中”,“百会”,“大椎”穴可以通过抑制神经细胞的炎症因子TNF-α以及级联反应执行凋亡蛋白酶Caspase-3,从而延缓了缺血损伤神经元的死亡,发挥对缺血性脑损伤的保护作用。
Objective:
Observe the effect of acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui of Resistance to injury and protection of the ischemic area. Observed ischemic brain tissue TNF-α and Caspase-3expression to explore acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui on the brain to produce the protective effect and possible mechanisms for this protective effect。
Methods:
SD male rats32were randomly divided into model group, sham operation group, DuiZhaoDian group and acupuncture group of four groups and each group of each only8.The establishment of a MCAO rat model, MCAO rat model of vital signs were stable after ZeaLonga score neurological deficit scores to verify the success of the model or not, six treatment of neurological function after the line with HE staining in rat brain tissue pathological changes in the flavor end labeling (TUNEL) assay detection of neuronal apoptosis, and apply the SP immunohistochemistry was detected in rat focal cerebral ischemia in brain tissue TNF-alpha and caspase-3positive cells.
Result:
1.Nerve functional score:(1) group compared with sham operation group, model group, DuiZhaoDian group, acupuncture group of neural function defect scale increased significantly, difference is very significant meaning (P<0.01);(2)group compared with model, DuiZhaoDian group, acupuncture group of neural function defect score lower, and the meridian group significantly different meaning (P<0.01);DuiZhaoDian group is significant difference(P<0.05);(3)group compared with DuiZhaoDian, acupuncture group of neural function defect scale reduced significantly, and significant difference(P<0.05).
2.Epidemic photochemical staining study dyeing:TNF alpha immune response results show that the number of positive cells group compared with sham operation group, model group DuiZhaoDian group of TNF alpha positive points cell count was higher, meaning with significant difference (P<0.01); Group compared with model, DuiZhaoDian group of TNF alpha positive points lower cell count, the difference was statistically significant (P<0.05), and the meridian group of TNF alpha positive cell count, reduce the significant difference (P<0.01); And DuiZhaoDian group compared, and the meridian group of TNF alpha positive cell count, reduce, the difference was statistically significant (P<0.05)
3.Image analysis ischemia side caspase-3immune response brain cell count and positive results:group compared with sham operation group, model group DuiZhaoDian group caspase-3points group was higher number of positive cells, meaning with significant difference (P<0.01); Group compared with model, and the meridian of the group Caspase-3positive cell count, reduce, the difference was statistically significant (P<0.01), DuiZhaoDian group of Caspase-3positive cells reduce the number is not obvious, meaningless differences (P>0.05); And DuiZhaoDian group compared, and the meridian group Caspase-three positive cell count, reduced significantly, and there is a significant difference of statistical significance (P<0.01).
Conclusion:
1. Ischemic damage rats obvious neural function defect signs, ischemia area nerve cells exist cell apoptosis, and acupuncture Du meridian people, Bai Hui, the Ren Zhong, the Da Zhui acupuncture point can obviously improve the rat nerve function and effectively restrain brain ischemia cell apoptosis happened after.
2. Acupuncture du meridian people, the Bai Hui, the Ren Zhong, the Da Zhui acupuncture point by inhibiting the neural cells inflammation factors TNF alpha and cascade reaction of the apoptotic protease expression of Caspase-3, delaying the ischemic injury the death of neurons, play to ischemic brain damage of protection.
引文
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