The Influence Of Jin’s Three Needle Treating HIBD Neonatal Rats In NO AndGSH-PX Expression

BackgroundNeonatal hypoxic ischemic encephalopathy (HIE) is a central neuronal damage because of asphyxia, hypoxia in perinatal period. Which is a serious complication of neonatal asphyxia, a critical condition, high fatality rate, it is also one of the important reasons of disability in children, appear some sequelaes after neurological injury, Such as cerebral palsy, mental retardation, ataxia, epilepsy, ADHD and so on. threat the Chinese babies in the life safety and body health seariously, but also brought great economic and spiritual suffering to the children’s family. Western medicine mainly use physical therapy, surgical treatment, drug treatment and rehabilitation means such as support or brace, improve the movement function of children with cerebral palsy. In recent years, jin’s three needle therapy academic system was developed by Professor Jin Rui, who was the chief professor for mentally handicapped children of Guangzhou University of Traditional Chinese Medicine, Showing the scene of unprecedented. But there is still a lack of mechanism of acupuncture curative effect to handicapped children on the research of experimental verification.ObjectiveIn recent years, jin’s three needle therapy academic system was developed by Professor Jin Rui, who was the chief professor for mentally handicapped children of Guangzhou University of Traditional Chinese Medicine, Showing the scene of unprecedented. But there is still a lack of mechanism of acupuncture curative effect to handicapped children on the research of experimental verification, Base of the modern medicine research, exploring the influence of using jin’s three needle on the content of NO and GSH-PX after acupuncturing HIBD rats, To verify whether the mechanism of action of neonatal hypoxic ischemia brain damage associated with the mitochondrial mediated apoptosis pathway in neural stem cells, And the stimulation of Jin San Zhen head point can inhibit the pathway, so as to provide a scientific evident for clinical treatment of Neonatal hypoxic ischemic encephalopathy children, which makes "Jin San Zhen" treatment of infantile encephalopathy become more mature, and provides a new effective method for the majority of medical staff on the treatment of HIBDMethodLiterature researchThis section is mainly to collect the analysis of etiology and pathogenesis, conclude the main treatment method associated with neonatal encephalopathy caused by intrauterine asphyxia. And the modern medicine about pathological research and treatment of neonatal encephalopathy caused by intrauterine asphyxia.Experimental researchThis section includes a total of two experimental parts.Experiment one HIBD neonatal rats, moldingEtablishing a HIBD pup rat model by a5min-delayed cesarean section which could simulate intrauterine asphyxia, the main operating is ligating bilateral uterine artery to21days pregnant rat, completely blocking blood supplying after5minutes to simulate the fetal intrauterine hypoxia ischemic brain injury model.selecting from the normal group and model group who comes from21days after the birth of the rats Randomly, remove the brain tissue after cervical dislocation, and HE staining to prove the molding was successful or not.Experiment two The influence of behavior, NO and GSH-PX concentration after acupuncturing to intrauterine distress HIBD rats.Putting natural birth rats for normal group; part of cesarean delivery delay and no acupuncture intervention were as the model group, cesarean delivery delay and acupuncturing as acupuncture group, according to different time point, we put Acupuncturing groups devided into nine groups, The point are14days,17days,20days,23days,26days,29days and32days,35days,38days, The period of each acupuncturing group rats received acupuncturing treatment for a week, All the newborn rats need behavioral detection in45days after the birth, including suspension test and slope test. Through behavior detection, Can reflect whether acupuncture on HIBD with the newborn rat neurons injury caused by ischemia hypoxia have repair effect.Adopting nitrate reductive enzymatic detecting the content of NO in rat cerebral cortex, because NO hasvarious induction effection in the process of poptosis of eural stem cells, can be directly damage DNA, inhibit DNA repair enzyme, and inhibition of the mitochondrial respiratory enzyme, promote cell energy failure, and then through the way of oxygen free radical damage induced apoptosis of neural stem cells. In addition, NO can promote the apoptosis inducing factor accumulation in the nucleus. Therefore, through the determination of the NO level in the rat brain cortex tissue to reflect the extent of the focal cerebral ischemia in rats of newborn neural cell apoptosis.We adopted DTNB method to detect GSH-PX (glutathione peroxidase) of the rat brain cortex, GSH-PX generally exists in the body, It is a kind of important peroxide decomposition enzyme, participating in the metabolism of oxygen free radicals in the body, combined with GSH and make it oxide reduction reaction specificly. When the rat cerebral ischemia hypoxia occurs, Lipid peroxide increases, Excess oxygen radicals fail to clear in time, they Will damage neurons and promote its apoptosis. GSH-PX enzyme to remove the body of excess toxic peroxide metabolites, it plays an important role for the protection of cell membrane structure and complete function. So by testing the GSH-PX levels, to reflect the ability of removal of oxygen free radicals, It explains the treatment of acupuncture mediated whether it has repairing the damaged neurons and inhibiting apoptosis.Conclusion:Document research1the understanding of the etiology and pathogenesis of infantile encephalopathy and clinical experience to the ancient traditional Chinese medicine is very rich, also provides a strong theoretical foundation to the encephalopathy and development of study on modern Chinese medicine treatment, thus have each breakthrough on medicine, acupuncture, massage, but lacks of standardized clinical treatment methods and curative effect evaluation method.2modern western medicine mainly study the pathogenesis of neonatal hypoxic ischemic encephalopathy and pathological physiological process, which main body is support treatment and symptomatic treatment, with hyperbaric oxygen, and low temperature, neurotrophic factor, inflammation inhibiting factor, methods such as neural stem cells transplantation, with appropriate rehabilitation training, but the overall curative effect and can’t get a satisfactory result.Experimental research1HIBD neonatal rats, moldingTake out of the newborn rat brain tissue after the birth of the21st day, and applying HE staining, the brain tissue slices shows a typical hypoxic ischemic brain damage under the microscopic, compared with the normal group, nerve cells are pathological morphological changed obviously, pyramidal cell shrinkage, the amount of nerve cells Decreased, and changed into a triangle, cytoplasm shows eosinophilic staining uniformly, the nuclear of cells are hyperchromatic, Structure disappeared, it indicates the models we made are successful.2behavior detectionIn terms of the behavior of these rats, suspension experiment results show that, the rats was born14,17,20days after accepting the suspension time acupuncture group was obviously longer than model group, the difference is statistically significant (P<0.05). And slope test results showed that the rats was born14,17,20,23days accept acupuncture group is shorter compared with model group, the difference has statistical significance (P<0.05). Above all, we know that early acupuncture intervention treatment is helpful to the newborn HIBD rat in behavior aspect., whose Pathological neurons due to ischemia anoxic injury.3NO detectionAdopting nitrate reductive enzymatic detecting the content of NO in rat cerebral cortex, Results showed that the model group was obviously higher than acupuncture group, The acupuncture group is lower than the normal group (P <0.05), the difference was statistically significant. The rats who accepted acupuncture from14to29day, compared with the rats who were acupunctured when they were32days, the content of NO was lower (p<0.05), and the difference was statistically significant, too. So the HIBD newborn rats received acupuncture as early as possible to reduce the brain’s nerve cells apoptosis significantly higher than the late accept and not accept acupuncture in rats.4GSH-PX detectionWe adopted disulfide generation2nitro benzoic acid method to detect the GSH-PX activity in rat cerebral cortex, Results showed that the acupuncture group and normal group, model group had obvious difference (P>0.05), The difference has no statistically significant; there was no obvious difference between acupuncture group and model group (P>0.05), the result was no statistically significant difference, the newborn rat GSH-PX content in Different acupuncture intervention time groups had no obvious difference (P>0.05). It may be explained acupuncture was efficient, but through other enzyme system, such as superoxide dismutase SOD, catalase CAT to remove excess oxygen radical in brain tissue, Rather than through GSH-PX ways to remove these toxic metabolite, to protect the damaged brain tissue.