Exploring The Effect Of Spared DRG, Acupuncture Spared DRG And Endogenetic C-fos, C-jun, GDNF, NT-4 On DRGn's Growing

Objective: To establish culture technology of adult cat's DRG in vitro and explore the effect of partial dorsal root rhizotomy and acupuncture on spared L6 DRG growth. The function of endogenous c-fos, c-jun, GDNF, NT-4 on DRG growth was also investigate so as to know the relation between DRG and the plasticity of spinal cord as well as acupuncture promoting plasticity of DRG and spinal cord.Methods: 10 adult cats were divided into 2 groups: normal control group;acupuncture spared DRG 7th day group (bilateral L1-L5, L7-S2 DRG were sectioned, but bilateral L6 DRG were spared; electro-needle stimulation was performed alternativly at unilateral two groups acupuncture point of including Zusanli and Xuanzhong , Futu and Sanyinjiao located in the distribution area of spinal nerve L6 DRG in operation side only one time everyday for 30 min for 7 days following bilateral partial root rhizotomy, animals surviving for 7 days after operation) . 5 cats were used in every group. Bilateral L6 DRGs of every group were taken out on the condition of asepsis and were cultured respectively in vitro. These adult cats' L6 DRGs' cells were planted equably into 48-well cultural plates coated by poly-L-Lysine and laminin, by the technology of vituline and equine serum culture in vitro after they were digested 3 hours by 0.25% collagenase. Medium of some of the acupuncture group's wells were totally replaced by each corresponding antibody-cultured medium, and others by DRGn-cultured medium at the 24th hour. After then, half of the medium were replaced every 2-3 days. Cultures were terminated after 7 days. Then, cultural cells were stained under the same condition using specific NSE (1:200) antibody, a neuron-specific marker, by the immunohistochemistry ABC method. The lengthof the neurites was measured by micro-measured ruler in upside-down light microscope. Data were analyzed by one-way anova and q test.Result: The results showed that most of the neurons have sticked to the bottom of the48-well plates and grew after being planted 10-14 hours.At the 1st day in the normal group: few neurons grew out neurites average length of neurite is 26.58?0.05um; the spared DRG group: half of neurons grew out neurites, the average length is 53.84?3.97um; the acupuncture group: most of neurons grew out neurites, the average length is 72.92+ 16.3 5 urn The neurites' length of the above mentioned three groups was distinct between every two groups (p<0.05), but there was not actual value because their standard deviation were too large.On the 3th day in the normal group: there were increasingly-growing cells and average neurite 's length is 56.66?16.52um; the spared DRG group: most of neurons grew out neurites, of which length is averagely 106.64+ 17.02um, and neural network began to be formed; the acupuncture group: all of neurons almost grew out neurites, of which average length is 133.56?15.67um, and neural network has begun to take shape. The average neurite's length of the above mentioned three groups is distinct between every two groups (p<0.05). But each antibody group's was similar with the acupuncture group's (p>0.05)On the 5th day. the normal group: Average neurite's length is 85.86?8.40um, and neural network began to be formed; the spared DRG group: Neurite length is averagely 106.64 17.02um, and neural network has been formed; The acupuncture group: Average neurite length is 133.56+15.67um, and neural network has been formed completely. The neurite's length of the above mentioned three groups was distinct between every two groups (p<0.05). Although average length of each antibody group's neurites was similar with the acupuncture group's (p>0.05), their neurites were shorter than the acupuncture'sOn the 7th day, the normal group; Average neurites' length is 1 06.64 14.99um, and neural network has been formed; the spared DRG group: Neurite length is averagely 1 55.36 15.16um, and neural network has been completely formed; the acupuncture group: Its' average neurite length is 205.44?19.76m, and neural network has been de...