Effect Of RAPA,3-MA And Acupuncture On Weight And MTOR Expression In HIBD Rats

Hypoxia-ischemic encephalopathy(HIE)is a Hypoxia-ischemic brain damage(HIBD)caused by hypoxia during perinatal period,this is the main reason of death and disability in newborn.Most of the surviving children suffered from neurological sequelae,such as learning disability,tic disorder,convulsions,memory deficits,slow development of motor system and cerebral palsy.A comprehensive treatment principle is applied because of its complicated pathogenesis in Hypoxia-ischemic encephalopathy(HIE).Feeding,increasing cerebral circulation,reducing intracranial pressure are the conventional treatment in acute phase;and rehabilitation exercises in recovery phase,respectively.With the advancements of Traditional Chinese Medicine,there is a breakthrough progress in treatment of Hypoxia-ischemic encephalopathy(HIE)like strengthening the internal organs,clearing the meridians,Xingnao Kai qiao.The main techniques include acupuncture,moxibustion,massage and Chinese medicine.Acupuncture is an important part of traditional Chinese medicine.It is widely accepted and applied with convenience and high efficiency.Studies have also found that acupuncture can effectively improve the symptoms of HIE sequelae such as growth disturbances,cerebral palsy,epilepsy etc.Studies proved that acupuncture not only improves some of the symptoms,but it also helps in regulating fumctions of organs.The earlier intervention with acupuncture,the greater the stimulation intensity and the better the outcome.Jin's three needles is a new treatment method summarized by Professor Jin Rui on the basis of the traditional acupuncture treatment.Professor Jin Rui often used three acupoints as a prescription in the treatment of diseases and the curative effect is remarkable.It becomes wellknown as "Jin's three needles" amongst his patients.Jin's three needles is a new representative genre in Lingnan acupuncture,Jin's three needles has been studied of encephalopathy in children in clinical practice.This study build the new HIBD rat model to investigate the pathological mechanism of Jin's three needles on HIE,to observe the changes of autophagy and the expression of mTOR in the hippocampus.ObjectiveEstablish new HIBD rat model to investigate the weight and mTOR expression in hippocampus of model group,acupuncture group,activator group,inhibitor group,acupuncture plus activator group and acupuncture plus inhibitor group.To explore the mechanism of Jin's three needles and to provide the exact theoretical basis for treatment of HIE.MethodologyEstablishment of animal model:HIBD rat model was established by intrauterine distress method,The experiment was divided into six groups:model group,acupuncture group,activator group,inhibitor group,acupuncture plus activator group,acupuncture plus inhibitor group.Eight subjects were assigned to each group.The interventions began on the 14th day,the acupuncture intervention once a day,strongly stimulated 10 times without needle retention,activator and inhibitor intervention to take intraperitoneal injection every day,all the intervention lasted for 14 days.Determination of experimental indicators:First:weighed and recorded on the 14th,21st and 28th day of the experiment respectively;Second:WB detection of hippocampal mTOR expression;Third:transmission electron microscopy observed the expression of autophagy in the hippocampus;ResultsFirst:There was no significant difference in weight between the experimental groups on the 14th day(p>0.05).On the 21st and the 28th day,the body weight of the activator group and the acupuncture plus activator group were significantly decreased compared with the model group(P<0.05).There was no statistically significant difference in body weight between acupuncture plus activator group and activator groups(p>0.05).Second:the WB showed that mTOR expression of the acupuncture group was significantly lower than that in the model group(P<0.05);there was no significant difference between activitor and model group(P>0.05);Compared with model group,mTOR expression was significantly decreased in inhibitor group.The difference was statistically significant(P<0.05);there was a statistically significantly lower between acupuncture plus activator group compared with the activator group(P<0.05);mTOR expression was significantly increased between the acupuncture plus inhibitor group compared with the inhibitor group,and the difference was statistically significant(P<0.05).Third:the experimental results showed that after administration of acupuncture,the number of autophagic bodies and the expression of autophagy increased.ConclusionFirst,acupuncture intervention is not a key factor to promote the weight,but also by other factors.Second,acupuncture intervention can regulate the expression of mTOR and increase the content of mTOR in hippocampus.Third,acupuncture can promote the expression of hippocampus autophagy,this mechanism may be completed with mTOR regulation.