| Neonatal hypoxia ischemic encephalopathy(hypoxia-ischemic encephalopathy,HIE)is the perinatal neonates craniocerebral injury caused by a lack of oxygen,is causing neonatal death and one of the important causes of neurological sequelae,survivors appeared more mental development is low,motor retardation,cerebral palsy,epilepsy and other neurological abnormalities.Therefore,the timely and effective treatment for children with HIE and his family is of great significance.In recent years,in the treatment of HIE,through the use ofacupuncture,moxibustion,acupoint injection,infantile massage,oralmedicine,such as fumigation treatment effect of traditional Chinese medicine,acupuncture,in particular,in the treatment of HIE sequela,curative effect is extremely significant.ObjectiveMitochondrial autophagy protection with BNIP3 mediates pathways as a breakthrough point,the successful building of HIBD rats jin’ s 3 needling head acupuncture point(brain three stitches,NieSanZhen,ZhiSanZhen)intervention,autophagy activation agent rapamycin at the same time,through the biological detection technology such as western blot observe autophagy related protein NIX(Nip3-like 19 kda protein X/adenovirus interacting protein 3-like),B cell lymphoma/leukemia-2(B cell lymphoma/leukemia-2,the Bcl-2)expression changes,aims to explore acupuncture on the damaged neurons induced by hypoxia ischemia mechanism and the influence of mitochondrial autophagy level,for the clinical treatment for neonatal hypoxia ischemic encephalopathy(hypoxia ischaemic encephalopathy,HIE)to provide new train of thought and basis.MethodsSPF SD rats,female(unfertilized)20,10 only males,according to 1:1 fold breeding,breeding success 16 pregnant rats,cesarean section 8 rats,rats only 8,cesarean section,after isoflurane anesthesia clamps bilateral uterine artery for 10 min,no more than 30 s,will be sterile gauze to cover on the bilateral uterine,clamps at the end of the time,take out the newborn rat,immediately,rescue,a total of 66 cubs mouse,the mouse out.According to random number table method,each group of 11,divided into model group,acupuncture group,activating agent,inhibitor,acupuncture plus activator group,acupuncture plus the suppression.Born in HIBD model rats started to intervene in the 14th day,intervention once a day,continuous intervention in 14 days.By observation of biological detection technology such as western blot autophagy related protein NIX,changes in the expression of Bcl-2,ResulisClamps after bilateral uterine artery manufacture by caesarean section that rat model of intrauterine distress HE staining results:the model group rats scattered arrangement of hippocampus nerve cell,visible swelling of neurons,loose structure,part of neurons in nucleus pycnosis,hyperchromatic and neuron cell vacuoles.Acupuncture group in the rat hippocampus nerve cells in the normal order,structure,clear nuclei,nucleoli.Inhibitor group structure of rat hippocampal CA1 area of nerve cells are destroyed is more serious,more variability necrosis nerve cells,the structure is loose,intercellular broadening,nucleus pycnosis,hyperchromatic,part of the nucleus neurons disappear,cavitation.The inhibitor group,acupuncture plus inhibitor groups of nerve cells in the hippocampus morphology damage slightly improved,cellular structure from the previous close,narrow gap,but neurons swelling phenomenon still exist,cell borders owe smooth.Instead,activator group in the rat hippocampus nerve cell density,nerve cell structure is complete,clear smooth cell borders.Acupuncture plus activator in the rat hippocampus neuron cytoplasm density increased,cell neatly,clear hierarchy.WB results model group offspring rat hippocampal tissue autophagy related protein expression of Bcl-2 significantly higher than the normal group,suggests building within 24 h of autophagy in higher degree,may be related to intrauterine distress model(lack of oxygen,ischemia)stimulate autophagy related proteins the Bcl-2 expression in great quantities,thus promotes the occurrence of autophagy.After building,NIX protein expression compared with model group,acupuncture group increased(P<0.05),suggesting acupuncture can promote autophagy appropriately,so as to promote the repair of brain cells.Compared with model group,NIX protein expression inhibitors group reduced(P<0.05),suggesting 3-MA can effectively inhibit cell mitochondria autophagy,on the basis of the inhibitor NIX the inhibitor of protein expression in acupuncture intervention group increased(P<0.05),suggesting the external conditions inhibit autophagy,"jin’s 3 needling" head acupuncture point acupuncture intervention can effectively promote cell autophagy.Gr.oup compared with model group,activator NIX protein expression increased(P<0.05),suggesting rapamycin can effectively promote cell mitochondria autophagy,on the basis of the activator in acupuncture intervention NIX protein expression increased in the activator group(P<0.05),suggesting "jin’ s 3 needling" head acupuncture point acupuncture intervention can cooperate to jointly promote cell autophagy activation agent.About the Bcl-2 expression,after building,compared with model group,acupuncture group of Bcl-2 protein expression decreased(P<0.05),prompted after given acupuncture intervention,mitochondrial autophagy is increased.Group compared with model group,inhibitor Bcl-2 protein expression increased(P<0.05),suggesting 3-MA can effectively inhibit cell mitochondria autophagy,on the basis of the inhibitors in acupuncture intervention the Bcl-2 protein expression the inhibitor group decreased(P<0.05),suggesting the external conditions inhibit autophagy,"jin’s 3 needling" head acupuncture point acupuncture intervention can effectively promote cell autophagy.Group compared with model group,activator Bcl-2 protein expression increase decrease(P<0.05),suggesting rapamycin can to a certain extent promote cell mitochondria autophagy,on the basis of the activator Bcl-2 protein expression in acupuncture intervention to reduce in the activator group(P<0.05),suggesting"jin’ s 3 needling" head acupuncture point acupuncture intervention can cooperate to jointly promote cell autophagy activation agent,due to ischemia anoxic damage nerves and brain cells have repair the positive role.Conclusion1.Acupuncture can strengthen NIX expression and at the same time promote cell autophagy.2.Acupuncture can promote autophagy related protein expression of Bcl-2,NIX,hints of acupuncture promoting tissue repair of damaged neurons may be associated with mitochondrial autophagy NIX、Bcl-2 mediated.3.Acupuncture can promote the expression of brain tissue damage mitochondrial autophagy,at the same time reduce the happening of the damaged area neurons apoptosis,acupuncture may by promoting moderate expression of mitochondrial autophagy,thereby reducing the occurrence of apoptosis. |
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