针刺对围产期缺氧缺血性脑损伤大鼠NGF、BDNF、GFAP及NO/NOS的影响
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摘要
第一部分文献综述
通过大量文献对关于围产期缺氧缺血性脑损伤的文献进行综合研究。首先,对祖国医学关于围产期缺氧缺血性脑损伤的病名、症状、病因病机、治疗等方面的进行了详细的分析;其次,着重从临床及实验研究方面对现代医学关于围产期缺氧缺血性脑损伤进行综述,引起HIBD的机制十分复杂,近年来,对其发病机理的研究不断深入,虽然HIBD的确切机理至今尚未完全澄清,但随着分子生物学研究的进展,从分子水平继续对新生儿及新生动物脑缺氧缺血作深入研究,必将为HIBD防治带来新的希望。目前尚且没有治疗儿童脑病的特效中西药,中外专家展开对该病的一系列研究,取得了一定成果。其中,针灸疗法日益引起国际的重视,有待进一步深入研究。
第二部分实验研究
目的:从半脑重量,脑组织NGF、BDGF、GFAP及血清NO、NOS方面探讨针刺疗法治疗新生大鼠围产期缺血缺氧性脑损伤的治疗作用及机制。
方法:分离并结扎孕鼠双侧子宫动脉,完全阻断血供25min娩出胎鼠,制成围产期缺血缺氧性脑损伤模型,成功的模型采取抛硬币法随机分入针刺组和模型组,每组各8只;正常组手术过程中仅用止血钳夹住孕鼠子宫颈,马上取出胎鼠,留取8只;其余胎鼠丢弃。正常组和模型组不治疗,针刺组于出生后7d后开始靳三针治疗,治疗至大鼠出生后30d。31d实验结束,将实验动物腹腔麻醉后快速断头取脑及血浆,检测脑组织NGF、BDGF、GFAP蛋白表达,及血清NO、NOS含量。
结果:
1.针刺对脑大体重量的影响
模型组半脑净重最低(0.467±0.021),较正常组(0.548±0.021)和针刺组(0.540±0.019)有显著性差异(P＜0.05);针刺组和正常组相比,差异无显著性差异(P＞0.05)。
2.针刺对脑组织NGF的影响
针刺组可显著提高大鼠脑组织NGF蛋白表达(3.90±0.47),与模型组(2.05±0.36)比较,差异有显著性(P＜0.01);模型组与正常组(1.12±0.24)比较,差异有显著性(P＜0.05)。
3.针刺对脑组织BDNF的影响
模型组(0.23±0.05)BDNF蛋白含量最低,模型组与正常组(0.50±0.21)比较,差异有显著性(P＜0.05);针刺组可显著提高大鼠脑组织BDNF蛋白的表达(1.24±0.18),与与模型组比较,差异有显著性(P＜0.01)。
4.针刺对脑组织GFAP的影响
正常大鼠脑组织GFAP呈弱表达(1.01±0.21),模型组GFAP的表达在三组中显著升高(1.63±0.19);针刺组(1.08±0.20)能显著降低GFAP蛋白的表达,与模型组差异显著(P＜0.05)。
5.针刺对血清NO及NOS的影响
模型组NO、NOS含量(87.65±55.75、43.37±15.97)较正常组(39.76±28.72、19.23±8.89)升高(P＜0.05),针刺组NO、NOS含量(60.79±51.42、29.19±12.89)较模型组下降(P＜0.05);针刺组和正常组相比,差异无显著性差异(P＞0.05)。
结论:1.围产期缺氧缺血对大鼠脑的生长发育将产生负面影响,而针刺疗法可提高发育期脑损伤大鼠脑大体的重量。2.针刺疗法显著提高了NGF活性,对受损变性的神经细胞有一定的恢复作用。3.针刺疗法可显著提高大鼠脑组织BDNF蛋白的表达,对受损变性的神经细胞有一定的恢复作用。4.针刺疗法能降低GFAP蛋白表达,避免胶质疤痕增生。5.针刺可降低围产期期脑损伤大鼠NO含量、NOS活性,对抗NO/NOS系统对神经系统的毒性作用。针刺疗法通过对NGF、BDNF、GFAP蛋白表达的影响,及对NO/NOS系统的调控作用,可能与护神经细胞有关,避免神经细胞死亡或凋亡有关;针刺疗法对上述各指标的调控,与其对缺血缺氧性脑病的治疗作用有关。
Part 1 Literature Review
We have comprehensively studied the literature on prenatal hypoxia-ischemic brain damage(HIBD) in a large scale.Firstly,analyze the term,symptoms,etiology,pathogenesis and treatments of prenatal HIBD in Chinese medicine;secondly,focus on clinical and experimental research on prenatal HIBD in modern medicine.The mechanism of HIBD is very complicated and further studies have been taken recent years.The pathogenesis of HIBD has still not been cleared up exactly,however,the progress on molecular biology assures the studies on HIBD at molecular level to be the newly hope as both the prevention and treatment to HIBD.No specific treatment has been found yet.Experts from home and aboard have carried out a series of research and obtained some achievements,in which,the acupuncture therapy has arisen more and more attention in the world and should be studied further.
PART 2 Experimental Researches
Objective:to study the effects and mechanism of acupuncture therapy in prenatal hypoxia-ischemic brain damage on newborn rats by measuring the weight of the hemisphere,the level of NGF,BDGF,GFAP in the brain tissue and NO,NOS in blood serum.
Methods:separated and ligated bilateral uteral arteries of pregnant rats to block the blood supply for 25 minutes then delivered fetal rats as the models of perinatal HIBD.16 qualified models were divided equally into acupuncture group and model group randomly by coins.The normal group had 8 fetal rats which were delivered immediately since the cervixes in pregnant rats were clipped by haemostatic forceps during operations only.The rest fetal rats were discarded.The normal group and model group received no treatment. The acupuncture group was treated by jin-3-needling from the7 day to 30 day postnatal.At the 31 day,when the experiment was over,cut the heads of the fetal rats under intraperitoneal anesthesia and removed the brains and plasma. Tested the protein expression of NGF,BDGF,GFAP in brain tissue and levels of NO,NOS in blood serum.
Results:
1.Effects of acupuncture on the weight of brain.
The net weight of hemisphere was the lowest in the model group(0.467±0.021),showing a significant difference against the normal group(0.548±0.021) and the acupuncture group(0.540±0.019)(P＜0.05);while there were no significant difference between the acupuncture group and the normal group (P＞0.05)
2.Effects of acupuncture on NGF in brain
The protein expression of NGF were increased evidently in the acupuncture group(3.90±0.47),whose had significant difference(P＜0.01) compared to the model group(2.05±0.36).And the difference between the model group and the normal group(1.12±0.24) was also significant(P＜0.05)
3.Effects of acupuncture on BDNF in brain
The protein levels of BDNF were the lowest in the model group(0.23±0.05),which had significant difference(P＜0.05) against the normal group (0.50±0.21);The protein expression of BDNF were increased evidently in the acupuncture group(1.24±0.18),whose had significant difference(P＜0.01) compared to the model group.
4.Effect of acupuncture on GFAP in brain
GFAP was weakly expressed in the normal brain tissue of rats(1.01±0.21),but increased obviously in the model group(1.63±0.19).In the acupuncture group,the expression of GFAP was decreased evidently(1.08±0.20),who had a significant difference against the model group(P＜0.05).
5.Effects of acupuncture on NO,NOS level in blood serum
The NO,NOS level in the model group(87.65±55.75、43.37±15.97) were higher than that in the normal group(39.76±28.72,19.23±8.89)(P＜0.05). The NO,NOS level in the acupuncture group(60.79±51.42、29.19±12.89) were decreased compared to the model group(P＜0.05).No significant difference was showed between the acupuncture group and the normal group(P＞0.05).
Conclusion:
1,Prenatal hypoxia ischemia has negative effects to the rat brain during its growth and development,while the acupuncture therapy can increase the weight of the injured brain in developmental stage.
2.The acupuncture therapy increases obviously the activity of NGF, which is responsible to the recovery of damaged neurons.
3.The acupuncture therapy increases obviously the expression of BDNF, which is also responsible to recover the damaged neurons.
4.The acupuncture therapy depresses the expression of the GFAP and avoids the proliferation of the neuroglial scar.
5.Acupuncture reduces the level of NO,the activity of NOS and their toxicity to the nerve system in the rats with prenatal brain damages.The effects of acupuncture on the expression of NGF,BDNF,GFAP and the regulation to the system of NO/NOS may be related to neuroprotection or by avoiding death or apoptosis of the neurons.
The regulation of the acupuncture therapy mentioned above reveals its effect on treating anoxic-ischemic encephalopathy.
通过大量文献对关于围产期缺氧缺血性脑损伤的文献进行综合研究。首先,对祖国医学关于围产期缺氧缺血性脑损伤的病名、症状、病因病机、治疗等方面的进行了详细的分析;其次,着重从临床及实验研究方面对现代医学关于围产期缺氧缺血性脑损伤进行综述,引起HIBD的机制十分复杂,近年来,对其发病机理的研究不断深入,虽然HIBD的确切机理至今尚未完全澄清,但随着分子生物学研究的进展,从分子水平继续对新生儿及新生动物脑缺氧缺血作深入研究,必将为HIBD防治带来新的希望。目前尚且没有治疗儿童脑病的特效中西药,中外专家展开对该病的一系列研究,取得了一定成果。其中,针灸疗法日益引起国际的重视,有待进一步深入研究。
第二部分实验研究
目的:从半脑重量,脑组织NGF、BDGF、GFAP及血清NO、NOS方面探讨针刺疗法治疗新生大鼠围产期缺血缺氧性脑损伤的治疗作用及机制。
方法:分离并结扎孕鼠双侧子宫动脉,完全阻断血供25min娩出胎鼠,制成围产期缺血缺氧性脑损伤模型,成功的模型采取抛硬币法随机分入针刺组和模型组,每组各8只;正常组手术过程中仅用止血钳夹住孕鼠子宫颈,马上取出胎鼠,留取8只;其余胎鼠丢弃。正常组和模型组不治疗,针刺组于出生后7d后开始靳三针治疗,治疗至大鼠出生后30d。31d实验结束,将实验动物腹腔麻醉后快速断头取脑及血浆,检测脑组织NGF、BDGF、GFAP蛋白表达,及血清NO、NOS含量。
结果:
1.针刺对脑大体重量的影响
模型组半脑净重最低(0.467±0.021),较正常组(0.548±0.021)和针刺组(0.540±0.019)有显著性差异(P＜0.05);针刺组和正常组相比,差异无显著性差异(P＞0.05)。
2.针刺对脑组织NGF的影响
针刺组可显著提高大鼠脑组织NGF蛋白表达(3.90±0.47),与模型组(2.05±0.36)比较,差异有显著性(P＜0.01);模型组与正常组(1.12±0.24)比较,差异有显著性(P＜0.05)。
3.针刺对脑组织BDNF的影响
模型组(0.23±0.05)BDNF蛋白含量最低,模型组与正常组(0.50±0.21)比较,差异有显著性(P＜0.05);针刺组可显著提高大鼠脑组织BDNF蛋白的表达(1.24±0.18),与与模型组比较,差异有显著性(P＜0.01)。
4.针刺对脑组织GFAP的影响
正常大鼠脑组织GFAP呈弱表达(1.01±0.21),模型组GFAP的表达在三组中显著升高(1.63±0.19);针刺组(1.08±0.20)能显著降低GFAP蛋白的表达,与模型组差异显著(P＜0.05)。
5.针刺对血清NO及NOS的影响
模型组NO、NOS含量(87.65±55.75、43.37±15.97)较正常组(39.76±28.72、19.23±8.89)升高(P＜0.05),针刺组NO、NOS含量(60.79±51.42、29.19±12.89)较模型组下降(P＜0.05);针刺组和正常组相比,差异无显著性差异(P＞0.05)。
结论:1.围产期缺氧缺血对大鼠脑的生长发育将产生负面影响,而针刺疗法可提高发育期脑损伤大鼠脑大体的重量。2.针刺疗法显著提高了NGF活性,对受损变性的神经细胞有一定的恢复作用。3.针刺疗法可显著提高大鼠脑组织BDNF蛋白的表达,对受损变性的神经细胞有一定的恢复作用。4.针刺疗法能降低GFAP蛋白表达,避免胶质疤痕增生。5.针刺可降低围产期期脑损伤大鼠NO含量、NOS活性,对抗NO/NOS系统对神经系统的毒性作用。针刺疗法通过对NGF、BDNF、GFAP蛋白表达的影响,及对NO/NOS系统的调控作用,可能与护神经细胞有关,避免神经细胞死亡或凋亡有关;针刺疗法对上述各指标的调控,与其对缺血缺氧性脑病的治疗作用有关。
Part 1 Literature Review
We have comprehensively studied the literature on prenatal hypoxia-ischemic brain damage(HIBD) in a large scale.Firstly,analyze the term,symptoms,etiology,pathogenesis and treatments of prenatal HIBD in Chinese medicine;secondly,focus on clinical and experimental research on prenatal HIBD in modern medicine.The mechanism of HIBD is very complicated and further studies have been taken recent years.The pathogenesis of HIBD has still not been cleared up exactly,however,the progress on molecular biology assures the studies on HIBD at molecular level to be the newly hope as both the prevention and treatment to HIBD.No specific treatment has been found yet.Experts from home and aboard have carried out a series of research and obtained some achievements,in which,the acupuncture therapy has arisen more and more attention in the world and should be studied further.
PART 2 Experimental Researches
Objective:to study the effects and mechanism of acupuncture therapy in prenatal hypoxia-ischemic brain damage on newborn rats by measuring the weight of the hemisphere,the level of NGF,BDGF,GFAP in the brain tissue and NO,NOS in blood serum.
Methods:separated and ligated bilateral uteral arteries of pregnant rats to block the blood supply for 25 minutes then delivered fetal rats as the models of perinatal HIBD.16 qualified models were divided equally into acupuncture group and model group randomly by coins.The normal group had 8 fetal rats which were delivered immediately since the cervixes in pregnant rats were clipped by haemostatic forceps during operations only.The rest fetal rats were discarded.The normal group and model group received no treatment. The acupuncture group was treated by jin-3-needling from the7 day to 30 day postnatal.At the 31 day,when the experiment was over,cut the heads of the fetal rats under intraperitoneal anesthesia and removed the brains and plasma. Tested the protein expression of NGF,BDGF,GFAP in brain tissue and levels of NO,NOS in blood serum.
Results:
1.Effects of acupuncture on the weight of brain.
The net weight of hemisphere was the lowest in the model group(0.467±0.021),showing a significant difference against the normal group(0.548±0.021) and the acupuncture group(0.540±0.019)(P＜0.05);while there were no significant difference between the acupuncture group and the normal group (P＞0.05)
2.Effects of acupuncture on NGF in brain
The protein expression of NGF were increased evidently in the acupuncture group(3.90±0.47),whose had significant difference(P＜0.01) compared to the model group(2.05±0.36).And the difference between the model group and the normal group(1.12±0.24) was also significant(P＜0.05)
3.Effects of acupuncture on BDNF in brain
The protein levels of BDNF were the lowest in the model group(0.23±0.05),which had significant difference(P＜0.05) against the normal group (0.50±0.21);The protein expression of BDNF were increased evidently in the acupuncture group(1.24±0.18),whose had significant difference(P＜0.01) compared to the model group.
4.Effect of acupuncture on GFAP in brain
GFAP was weakly expressed in the normal brain tissue of rats(1.01±0.21),but increased obviously in the model group(1.63±0.19).In the acupuncture group,the expression of GFAP was decreased evidently(1.08±0.20),who had a significant difference against the model group(P＜0.05).
5.Effects of acupuncture on NO,NOS level in blood serum
The NO,NOS level in the model group(87.65±55.75、43.37±15.97) were higher than that in the normal group(39.76±28.72,19.23±8.89)(P＜0.05). The NO,NOS level in the acupuncture group(60.79±51.42、29.19±12.89) were decreased compared to the model group(P＜0.05).No significant difference was showed between the acupuncture group and the normal group(P＞0.05).
Conclusion:
1,Prenatal hypoxia ischemia has negative effects to the rat brain during its growth and development,while the acupuncture therapy can increase the weight of the injured brain in developmental stage.
2.The acupuncture therapy increases obviously the activity of NGF, which is responsible to the recovery of damaged neurons.
3.The acupuncture therapy increases obviously the expression of BDNF, which is also responsible to recover the damaged neurons.
4.The acupuncture therapy depresses the expression of the GFAP and avoids the proliferation of the neuroglial scar.
5.Acupuncture reduces the level of NO,the activity of NOS and their toxicity to the nerve system in the rats with prenatal brain damages.The effects of acupuncture on the expression of NGF,BDNF,GFAP and the regulation to the system of NO/NOS may be related to neuroprotection or by avoiding death or apoptosis of the neurons.
The regulation of the acupuncture therapy mentioned above reveals its effect on treating anoxic-ischemic encephalopathy.
引文
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