| Purpose:This experiment chooses the model of the rat sciatic nerve chronic damage,with the small animals Vevo2100 high resolution ultrasound image.acupuncture “Huantiao”(gb30 point) under the different tissue,the muscle tissue and the nerve trunk.From the perspective of acupuncture depth,this paper discusses the goal of acupuncture with the different part of the relationship between acupuncture effect.And through the research on deep stab “Huantiao” on the nerve injury in rats and the effect of L4-L5 spinal cord of rats and dorsal root ganglion area “PI3K”,“AKT” expression,further analysis of deep stab“Huantiao” during the repair of sciatic nerve injury effect and the related signal pathways PI3K-AKT.Material and method:This experiment chooses healthy SD rats,a total of 48,according to the random number table distribution method were an average of 48 rats can be divided into four groups, each group of 12. Blank control group, model group, deep stab, shallow group.Building methods according to the Mackinnon design of silicone tube extrusion into the rat sciatic nerve chronic damage model. Blank group: under the same conditions, do not do other processing,maintain normal breeding.Model group:under the same conditions,do not do any processing, maintain normal breeding. Deep stab group: 14 d after the success of the building, in the high frequency of animal ultrasound guided needle “ Huantiao ”, under the ultrasonic probe, from between the ischial tuberosity and the femoral greater trochanter of the needle into the sag to the direction of the sciatic nerve acupuncture,timely adjust the direction, observation of the nerve stem tip to touch,the instant ultrasonographic in around the nerve trunk muscle twitching, toes trembling,Record the depth of the deep stab of 15 to 17 mm. Connection electric acupuncture treatment,the other end with medical adhesive binding to the rat tail made irrelevant the acupuncture needle.Choose density wave,current,frequency of 2HZ,cupping treatment when the strength of the mild local twitch acupuncture parts shall prevail,each treatment for 15 min.1/d,14 d continuous treatment.Shallow thorn group:after the success of the building d,14 in the high frequency small animals by ultrasound guiding left sciatic nerve acupuncture,to observe the needle point hit a muscle layer of about 5mm,Ultrasonic ultrasonographic show tip does not touch the nerve trunk.Connection electric acupuncture treatment,the other end with medical adhesive binding to the rat tail made irrelevant the acupuncture needle.Choose density wave,current,frequency of 2HZ,cupping treatment when the strength of the mild local twitch acupuncture parts shall prevail,each treatment for 15 min.1/d,14 d continuous treatment.14 d after the treatment,in small animals high frequency ultrasound guidance,observation of sciatic nerve internal echo and observed the pathological morphological changes of sciatic nerve,the nerve diameter is measured from the ultrasound images.By detecting the rat sciatic nerve function index to evaluate each rat muscle strength and motor function recovery;Measurement neural electrophysiology rats nerve conduction velocity evaluation of nerve function defect recovery.Death by excessive anesthesia method in rats,the rat sciatic nerve injury section and L4-5 segmental spinal cord and dorsal root ganglion for index detection.BY HE staining observation of sciatic nerve injury in the pathological change of the form.Apoptosis by TUNEL method to observe the sciatic nerve lesions.Immunohisto chemistry and Western blot to the observe rat L4-5 segmental spinal cord and dorsal root gaglion area “PI3K”,“AKT”expression intensity,the results were statistically analyzed.Results: 1. The behavior observed in rats Rats postoperative mental state is good, the skin color gloss, normal diet, physical activity appear limp, and rear limb weakness.Digits show a slight valgus toe disfunction and vein, and autophagy.Treatment group rats, by contrast, the muscles of the lower limb activity increased significantly, the treatment group is better than the model group, deep stab group is better than that of shallow thorn. 2. The rat sciatic nerve function index Rat sciatic nerve function index(SFI) showed that after building, SFI index of model group was obviously lower than that of blank group(P < 0.05), suggesting the rat made mould of motor function decline and success.Treatment group compared with model group, treatment group of SFI index increased significantly(P <0.05).Deep stab stab and shallow group compared with deep stab SFI index of treatment group was obviously higher than that of shallow thorn SFI index of treatment group(P < 0.05). 3. The nerve electrophysiology Nerve conduction velocity(MNCV) showed that, after building model group compared with the blank group, model group MNCV lower(P < 0.05), suggesting building success.Model group and treatment group MNCV values there are significant differences(P < 0.05), treatment group nerve conduction velocity is higher than the model group.Deep stab stab and shallow group, compared than deep stab nerve conduction velocity is higher than shallow thorn group(P < 0.05). 4. High frequency small animal ultrasound image analysis Under high frequency ultrasound imaging, the use of silicone tube strips strong echo recognition conformity.Model group nerve press-fitting section and two end diameter increases gradually, part formed the tumor sample change.Conformity of areas appear different degree of demyelinating changes, nerve fibers appear edema, the swelling of axons.Epineurium thickening, edge neat.Treatment group can be seen in the silicone tube strong linear echo, appear edema, but keep good continuity, nerve fibers demyelination changes get better inhibition, did not see epineurium thickening, edge is neat, no tumor sample change.Pathological changes of the deep stab group improved significantly better than light group. 5. Each group rats sciatic nerve HE dyeing observation Blank group belongs to the normal nerve fibers and axon, nerve sheath film, neat schwann cells, the structure is clear.Model group direction, nerve fiber axon disappear, myelin Waller degeneration occurs, collapse and disappear, schwann cells increased.The treatment group compared with model group, treatment group nerve fibers reduced mussily, did not see myelin appear only depigmentation disintegration, schwann cells increased, visible newborn nerve fibers.Deep stab group improved the pathological changes of superior to shallow thorn. 6. TUNEL apoptosis detection Mark apoptotic cells by TUNEL method, found that apoptotic cells mainly appeared in the sciatic nerve injury, the lesions were almost no apoptotic cells appeared.The perspective of model group can be found in a number of apoptotic cells.The apoptotic cells and the treatment group compared with model group, mainly distributed in sciatic nerve injury, number of apoptosis cells treatment group were less than model group(P < 0.05).The number of apoptosis cells deep stab group was obviously lower shallow thorn group of apoptotic cell count(P < 0.05). 7. Each group in the rat spinal cord L4- L5 "PI3- K", "AKT protein expression level of detection Use of immunohistochemistry and Western blot detection "PI3- K", "AKT" expression.Results show that compared with the blank group, each group of L4- L5 segmental spinal cord of rats and in the dorsal root ganglion "PI3- K" and "AKT protein expression were significantly higher(P < 0.05);Treatment group compared with model group, treatment group rats L4, L5 segmental spinal cord and dorsal root ganglion "PI3- K" and "AKT protein expression were significantly higher(P < 0.05);Deep stab stab and shallow group compared with deep stab group of L4- L5 segmental spinal cord of rats and in the dorsal root ganglion "PI3- K" and "AKT protein expression was higher than shallow" thorn group(P < 0.05).Conclusion: 1. Acupuncture can improve rats sciatic nerve injury recovery of neurological function, promote muscle recovery strength, prevent muscle atrophy, improve the recovery of motor function in rats behavior.Deep stab thorn group is superior to the shallow group. 2. Acupuncture can improve the pathological changes of damaged nerve fiber nerve conduction velocity, accelerate the regeneration of nerve fibers.Deep stab thorn group recovery than shallow group. 3. Acupuncture can increase the PI3- K ", " AKT protein expression and activation of PI3K-AKT signaling pathways that inhibit the apoptosis of nerve cells.Deep stab thorn group is superior to the shallow group. 4. The activation of PI3K- AKT signaling pathway inhibition of cell apoptosis may be a deep stab "gb 30" repair one of the important mechanisms of sciatic nerve injury. |
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