Influence Of Acupunture On Study-memory And Levels Of Influanammatory Mediators Protein Expression Of Young Rat Model With CP

Purpose:Construct hypoxic ischemic brain injury models, observed behavioral changes in different groups of mice with cerebral palsy, brain tissue and brain tissue pathological changes in inflammatory mediators(TNF-αIL-6) protein expression.Study Methods:1. Materials and ConditionsAnimals:7d old newborn SD mouse, supplied by the Guangzhou University of Traditional Chinese Medicine Laboratory Animal Center, animal quality certification number:0043108; license number:SCXK (Guangdong) 2008-0020. 8 package 90 numbers, required to reach> 12g within 7 days, mice were breed in clean laboratory having relative humidity 60% to 80% and equipped with fluorescent light, regular ventilation.Animal Laboratory:Animal Centre in Guangzhou University of Traditional Chinese Medicine, clean and controlled environment with temperature 22℃～26℃, relative humidity 60～80%, regular ventilation; animals facilities No.0023091; license no.:0019777.Animal Feeding:All the animals were feed in the Animal Center of Guangzhou University of Traditional Chinese Medicine; caged in groups, fed by conventional standard mouse pellets (containing protein 0.23g, d Fat 0.047g, sodium 0.0024g), from Beijing Feed Co., Ltd. Cooperation Section Australian production.2. models constructionAfter ether (alcohol) inhalation and under anesthesia, seperate the left carotid artery ligation, the incision was sutured back into the cage for 2h, then put them under a constant temperature water bath at 37℃in sealed containers; apply 1L/min speed injecting 8% oxygen + 92% nitrogen gas mixture. Mice were taken out 2.5h later for testing after another 1h for behaviour testing. Those which cannot turn round, and those have balancing abnormalities, and/or left turning are considered as successful models; they were then randomly divided into 5 groups and put back to the cage. The sham group, after anesthesia were having the left carotid artery separated but not ligation; after the surgery they were not tested under hypoxia, behavior tested being no significancy.3. grouping and treatment:(1) acupuncture group, all mice treated with acupuncture needles to limbs and head points using 2mm diameter 20mm long acupuncture needles, first at Neiguan and Yongquan o little bleeding and do not leave needles; then at limb acupoints Quchi and Zusanli for 2mm, the head points Baihui, temporalⅠwere inserted at level after feeling the needle being tight, stay there for 10min.(2) moxibustion group, apply mild smoke-free moxibustion to Governor Vessel for 10 minutes.(3) acupuncture+moxibustion group, after acupuncture treatment apply mild moxibustion Governor Vessel for 10 minutes.(4) normal group, sham group, model group did not give any treatment.Location Method:Point Positioning reference "experimental acupuncture" [2], combined with human and animal bones degree analogy:parietal bone center will Baihui; external ear canal, straight 0.8cm simulate take'Temporal 1 needle'; radial bone proximal lateral aspect from the wrist off 3cm or so take'Neiguan'; anterior aspect of lower leg under the fibular head and about 5mm take'Zusanli'; hind legs before the middle of the sole take'Yongquan' parietal bone to the coccyx as the center of the Governor meridian.The timing of intervention:the treatment group after 1,2 d (after birth 8,9 d) rest, after the first phase treatment of 3-8d (born 10-15d) for 6 days, 9d (born 16d)rest, after the second phase treatment 10-15d (born 17-22d) for 6 days, after 18d (born 25d) were sacrificed.4, using hematoxylin an eosin (H. E) staining and pathological brain tissue, reflecting hypoxic-ischemic brain tissue pathological changes.5, by radioimmunoassay of serum TNF-a, IL-6 levels. Results:1. Weight There was no difference among 6 groups on the first day. Mean number of gaining weights among 6 groups were different or incongruent after 18 days which was significantly(P<0.01).No statistic difference were reveled among the gaining trend of weights among the①normal group,②Sham operation group,⑤moxibustion group and⑥acu-moxibustion group, as well as between the③model group,④acupunture group. While the differences between the former 4groups and the latter 2 groups showed statistical significance on the gaining trend of weights(P=0.000<0.01).Muscle strength (overhanging test):②Sham operation group>①normal group>⑥acu-moxibustion group>⑤moxibustion group>③model group>④acupunture groupBalance (inclined plane test):①normal group>(2)Sham operation group>⑥acu-moxibustion group>⑤moxibustion group>④acupunture group>③model group.The variated increases of Muscle strength(overhanging test) and Balance (inclined plane test) produced by different treatments were statistical significance (P=0.000<0.01)Intelligence and memory (Y-maze test):①ormal group>②Sham operation group>⑥acu-moxibustion group>⑤moxibustion group>④acupunture group>③model group. Correct times were significant different(P=0.000<0.01)2. Histopathology of brain tissueNormal neuron number Compared to model group, the nomal neuron number in acupunture group,moxibusiton group and acu-moxi group raised significantly after treatment (P<0.05). While no obvious differences were found among these 3 groups. The normal neurons of the Sham operation group was lower than the normal group, but not statistically related (P>0.05)3. Levels of IL-6 and TNF-αLevels of IL-6 in acupunture group,moxibusiton group and acu-moxi group were elevated then the model group after treatment, which showed significantly difference (P<0.05).The normal group (P<0.01) and Sham operation group (P>0.05) had higher level then the model group, however, neither of then showed significant difference.Compared to the model group,levels of Levels of IL-6 in acupunture group,moxibusiton group and acu-moxi group were decreased after treatment, which showed significantly difference (P<0.05).The normal group (P<0.01) and Sham operation group(P>0.05) had lower level then the model group, however, neither of then showed significant difference.Levels of TNF-αin acupunture group, moxibusiton group and acu-moxi group were elevated then the model group after treatment, which showed significantly difference (P<0.05).The normal group (P<0.01) and Sham operation group (P>0.05) had higher level then the model group, however, neither of then showed significant difference.Compared to the model group, levels of TNF-αin acupunture group, moxibusiton group and acu-moxi group were decreased after treatment, which showed significantly difference (P<0.05).The normal group (P<0.01) and Sham operation group (P>0.05) had lower level then the model group, however, neither of then showed significant difference.Conelusion:1. Carotid artery ligation and hypoxia can successfully simulate the hypoxic ischemic brain injury model.2. Acupuncture can help improve behavior juvenile cerebral palsy, strength, intelligence and memory, the acupuncture and moxibustion on promoting the role of both CP mice, but combined the two will perform the best.3. Acupuncture can improve cerebral palsy mice nerve cell morphology and increase the hippocampus of normal nerve cell counts.4. Acupuncture has effect on inflammatory factors of CP mice and can modulae the increased inflammatory factors...