The Study About The Mechanism Which Electroacupuncture At Jiaji Points ?EX-B2? Protects BDNF Of Lumbar Spinal Neurons In ALS-SOD1^G93ATransgenic Mice

Objective:ALS-SOD1^G93A transgenic mice as experimental object,through the clip ridge electricity for survival in mice,the onset time and the influence of the behavior,and from the effects of BDNF as the breakthrough point,to study waist clip ridge electricity for mice marrow anterior horn the effects of BDNF expression,provide experimental basis for clip ridge electric acupuncture treatment of ALS.Methods:The first part of clip ridge electricity for ALS-SOD1^G93A transgenic mice survival,infection time and the influence of the behavior:1.Identification of ALS-SOD1^G93A transgenic mice:ALS-SOD1^G93A Transgenic mice as the experimental subjects,in the days of age 30 days to apply the Jackson laboratory provided by primer PCR identification of sequence and scheme,according to the characters of disease progression in mice to observe its phenotype.2.Grouping and disposal:24 ALS-SOD1^G93A transgenic mice,according to random number table method were randomly divided into electric acupuncture group,the hand needle group and model group 8,day age 60 days began to intervene,model group shall be bound to a fixed;Hand needle group on the basis of model group mice to bilateral L1?2,L5?6 clip ridge point acupuncture intervention;Electric acupuncture group in hand needle group,on the basis of the torso ipsilateral cupping treatment,frequency of 2 Hz.20 min/times,2 times per week,a total of 4 weeks.3.Nerve function score and behavioral testing:joint application turn tail suspension test,weighing,rod experiments to observe the onset and progress of the disease in mice.The second part of the waist clip ridge will raise ALS-SOD1^G93A transgenic mice marrow of anterior horn of BDNF mechanism research:1.Grouping and disposal:24 ALS-SOD1^G93A transgenic mice;6 wild-type mice with litter,according to random number table method 24 transgenic mice random divided into electric acupuncture group,the hand needle group,model group 6,6 wild-type mice with litter as a negative control,electric acupuncture group,the hand needle group,model group,treatment with the first part,the negative control group without any treatment.2.Nissl's staining:observing the type 120 days day age waist marrow of anterior horn motor neurons morphology.3.Immunohistochemical method:to test the type 120-day day age waist marrow anterior horn the expression of BDNF.Results:1.Phenotypic characterization:ALS-SOD1^G93A transgenic mice genetically engineered mice day age 90 days or so,tail suspension test in hind limbs tremor or unable to extend,100-120 days or so slow and dragging the hind legs,150 days to complete paralysis in mice,achieve lifetime end point.2.Analysis of weight:compared with model group,the hand needle group has a less weight loss in mice,but no statistical significance?P>0.05?,the curative group can obviously delay the mice body weight loss,there is significant difference?P<0.05?;Compared with hand needle group,the curative group of weight loss in mice have been delayed,but there was no significant difference?P<0.05?.3.rotarod test:from the 19 weeks to 21 weeks,compared with model group,the hand of mice needle rod movement function significantly improved?P<0.05?,the curative group improved more significantly?P<0.01?.4.Analysis of onset and survival:compared with model group,the curative group of mice significantly delay the onset time,with significant difference?P>0.05?,hand needle group also happen delay onset time of mice,but there was no significant difference?P<0.05?.Compared with model group,the hand needle group stage was extended,the survival of mice with significant difference?P<0.05?,the curative group survival extend more obvious,there are significant differences?P<0.01?;Compared with hand needle group,extended survival in mice by 6 days electric acupuncture group,with significant difference?P<0.05?.5.Morphology:he's staining to observe waist marrow number of anterior horn motor neurons in mice and morphology,negative control group waist marrow anterior horn neurons contains rich nissl body,nerve cells form clear;Model group lumbar anterior horn pulposus obviously loose edema in mice,cavitation,structure of nerve cells are destroyed,visible in the cell body blue irregular lump,austenite amount decreased significantly,the decrease in the number of motor neurons?P<0.01?;Hand needle and electric acupuncture group compared with model group number waist marrow of anterior horn motor cells of mice increased?P<0.05,P<0.01?,the curative group hand needle group increased more obviously?P<0.05?,the hand needle and electric acupuncture group waist marrow in the anterior horn motor neurons in mice,austenite increase obviously.6.Waist clip ridge electricity for ALS-SOD1^G93A transgenic mice marrow anterior horn the expression of BDNF:the influence of immunohistochemical staining,according to the results of model group mice waist marrow anterior horn the expression of BDNF declined significantly?P<0.01?;Compared with model group,the expression of BDNF hand needle group and the curative group was obviously increased?P<0.01?,and electric acupuncture is better than the hand needle group?P<0.05?.Conclusion:1.Electroacupuncture at jiaji points?EX-B2?can improve motor function,delay the onset of disease and prolong survival time in ALS-SOD1^G93A transgenic mice,and the effect is better than acupuncture.2.Electroacupuncture at jiaji points?EX-B2?can improve ALS-SOD1^G93A transgenic mice waist marrow of anterior horn motor neurons morphology changes.3.Electroacupuncture at jiaji points?EX-B2?may by raising ALS-SOD1^G93A transgenic mice waist marrow anterior horn the expression of BDNF,play a role of protection of neurons.