本文選題：坐骨神經(jīng)損傷 + 不同頻率��； 參考：《遼寧中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的:通過比較不同頻率(2Hz,100Hz)電針對坐骨神經(jīng)鉗夾損傷大鼠的實驗觀察,以“Bcl-2”“P53”“Bax”作為檢測指標,揭示電針治療坐骨神經(jīng)損傷抗凋亡作用的機制,探求周圍神經(jīng)損傷后抑制神經(jīng)元凋亡的最佳治療頻率。材料與方法:參考鉗夾法制作模型。選用SPF級SD大鼠48只,體重250g-350g,7-9周,雌雄各半。采用隨機數(shù)字表分配法分為四組:空白組、模型組、治療組1(2Hz)、治療組2(100Hz),每組各12只�？瞻捉M:正常飼養(yǎng),將動物固定在自制的塑料筒內(nèi),鼠尾和兩后肢露在筒外固定,持續(xù)15min,不做任何治療。模型組:造模成功后,將動物固定在自制的塑料筒內(nèi),鼠尾和兩后肢露在筒外固定,持續(xù)15min,不做任何治療。治療組1:造模成功后,于造模第8天深刺“環(huán)跳”穴。連接電針儀進行治療,另一端制成無關(guān)電極,選用頻率為2Hz的連續(xù)波,電針治療強度以針刺部位局部輕微抽動為準,每次治療15min,1次/天,連續(xù)治療14天。治療組2頻率為100Hz,除頻率與治療組1不同外,其余予相同處置。各組在造模第8天、22天進行足底印跡檢查。取損傷處坐骨神經(jīng)鉗夾處約上下2mm的坐骨神經(jīng)及同側(cè)L4-L5節(jié)段脊髓,用來制作成切片,通過HE染色,Western blot免疫組化等方法測定神經(jīng)細胞凋亡相關(guān)蛋白Bcl-2、P53及Bax的表達。結(jié)果:1.一般狀態(tài)觀察攝食飲水及大小便正常,造模后無感染;除空白組其余各組呈現(xiàn)拖趾狀或蹦跳狀步態(tài),各組間差異不明顯。治療組1和治療組2在造模第15天步態(tài)開始恢復(fù)。2.坐骨神經(jīng)功能指數(shù)(SFI)測定治療組與模型組相比SFI值升高(P0.01),且治療組1優(yōu)于治療組2(P0.01)。3.HE染色模型組神經(jīng)纖維排列呈不規(guī)則、紊亂的形式,且神經(jīng)纖維周圍的雪旺細胞增加,軸突和髓鞘發(fā)生裂解,幾乎完全消失。治療組神經(jīng)纖維病理變化較輕,神經(jīng)纖維出現(xiàn)新的生長,兩者對比治療組1較輕。4.Western blot定性、定量檢測4.1 Western blot Bcl-2檢測結(jié)果:四組實驗中bcl-2蛋白均有明顯條帶,測定灰度值,(1)與空白組比較,模型組、治療組1、治療組2中bcl-2蛋白表達明顯升高(P0.01);(2)與模型組相比,治療組1和治療組2中bcl-2蛋白表達明顯升高(P0.01);(3)與治療組1相比,治療組2中bcl-2蛋白表達降低(P0.05)。4.2 Western blot Bax檢測結(jié)果:四組實驗中Bax蛋白均有明顯條帶,測定的灰度值,(1)與空白組相比,模型組和治療組2中Bax蛋白表達明顯升高(P0.01),治療組1中Bax蛋白表達升高(P0.05);(2)與模型組相比,治療組1和治療組2中Bax表達明顯降低(P0.01);(3)與治療組1相比,治療組2中Bax表達升高(P0.05)。4.3 Western blot P53檢測結(jié)果:四組實驗中P53均有明顯條帶,測定的灰度值,(1)與空白組相比,模型組和治療組2中P53蛋白表達明顯升高(P0.01),治療組1中P53蛋白表達升高(P0.05);;(2)與模型組相比,治療組1和治療組2中P53蛋白表達明顯降低(P0.01);(3)與治療組1相比,治療組2中P53蛋白表達升高(P0.05)。5.免疫組織化檢測結(jié)果5.1 Bcl-2免疫組織化檢測結(jié)果:四組實驗中bcl-2蛋白均有表達,(1)與空白組相比,模型組中bcl-2蛋白表達升高(P0.05),治療組1與治療組2中bcl-2蛋白表達明顯升高(P0.01);(2)與模型組相比,治療組1中bcl-2蛋白表達明顯升高(P0.01),治療組2中bcl-2蛋白表達升高(P0.05);(3)與治療組1相比,治療組2中bcl-2蛋白表達升高(P0.05)。5.2 Bax免疫組織化檢測結(jié)果:四組實驗中Bax蛋白均有表達,(1)與空白相比,模型組和治療組2中Bax蛋白表達明顯升高(P0.01),治療組1中Bax表達均升高(P0.05);(2)與模型組相比,治療組1和治療組2中Bax蛋白表達明顯降低(P0.01);(3)與治療組1相比,治療組2中Bax蛋白表達升高(P0.05)。5.3 P53免疫組織化檢測結(jié)果:四組實驗中P53均有表達,(1)與空白組相比,模型組和治療組2中P53表達明顯升高(P0.01),治療組1中P53表達升高(P0.05);(2)與模型組相比,治療組1和治療組2中P53表達明顯降低(P0.01);(3)與治療組1相比,治療組2中P53表達升高(P0.05)。結(jié)論:1.電針可以改善急性坐骨神經(jīng)損傷大鼠坐骨神經(jīng)運動功能的恢復(fù),且2Hz組優(yōu)于100Hz組。2.電針可調(diào)節(jié)急性坐骨神經(jīng)鉗夾損傷后脊髓前角細胞運動神經(jīng)元凋亡及相關(guān)蛋白Bcl-2、bax及p53的表達,機制可能與增強抗凋亡作用因子Bcl-2,降低促凋亡因子Bax、P53的表達有關(guān)。3.電針可調(diào)節(jié)急性坐骨神經(jīng)鉗夾損傷后脊髓前角細胞運動神經(jīng)元凋亡及相關(guān)蛋白Bcl-2、bax及p53的表達,頻率2Hz優(yōu)于100Hz。
[Abstract]:Objective: To compare the experimental observation of sciatic nerve clamp injury in rats with different frequency (2Hz, 100Hz) electroacupuncture, with "Bcl-2" "P53" "Bax" as the detection index, to reveal the mechanism of anti apoptosis by electroacupuncture in the treatment of sciatic nerve injury, and to explore the best treatment frequency for the inhibition of neuron apoptosis after peripheral nerve injury. The model was made by clamp method. 48 SPF class SD rats were selected, weight 250g-350g, 7-9 weeks, and male and female. The random digital table distribution method was divided into four groups: blank group, model group, treatment group 1 (2Hz), treatment group 2 (100Hz), each group was 12. The blank group was kept in the homemade plastic tube, the rat tail and two hind limbs were fixed outside the tube. Continuous 15min, without any treatment. Model group: after the model was successful, the animal was fixed in the homemade plastic tube, the rat tail and the two hind limbs were exposed to the tube and fixed outside the tube. 15min was sustained without any treatment. After the successful model of 1: in the treatment group, the "ring jump" point was deeply prickled for eighth days. The electroacupuncture instrument was connected to the treatment, the other end was made into the independent electrode and selection frequency. For the continuous wave of 2Hz, the strength of the electroacupuncture treatment was based on the local mild aspiration of the acupuncture site. Each time was treated with 15min, 1 times / day and continuous treatment for 14 days. The 2 frequency of the treatment group was 100Hz, except for the frequency of 1 different from the treatment group. The same disposal was given in the treatment group for eighth days and 22 days. The sciatic nerve clamp at the injury site was about 2mm The sciatic nerve and the L4-L5 segmental spinal cord of the same side were used to make slices. The expression of Bcl-2, P53 and Bax of neuronal apoptosis related proteins was measured by HE staining and Western blot immunohistochemical method. Results: 1. the normal state of drinking water and urine and stool were observed in general state, and no infection after the model was made; the other groups in the blank group showed toes or jumping. In the treatment group 1 and the treatment group 2, the SFI value of the.2. sciatic nerve function index (SFI) in the treatment group was increased (P0.01) compared with the model group (P0.01), and the treatment group was better than the treatment group 2 (P0.01).3.HE staining model group with irregular, irregular form, and the nerve fiber week. The Schwann cells in the circumference were increased, the axon and myelin sheath were cracked and almost completely disappeared. The pathological changes of the nerve fibers in the treatment group were lighter, and the nerve fibers appeared new growth. The comparison between the two groups was 1 light.4.Western blot, and the quantitative detection of the results of 4.1 Western blot Bcl-2 detection: the bcl-2 protein in the four groups had obvious bands, and the gray level was measured. Value, (1) compared with the blank group, the model group, the treatment group 1, the bcl-2 protein expression in the treatment group was significantly increased (P0.01); (2) the expression of Bcl-2 protein in the treatment group 1 and the treatment group was significantly higher than that in the model group (P0.01); (3) compared with the treatment group 1, the bcl-2 protein expression decreased (P0.05).4.2 Western blot Bax detection results in the treatment group 2: four group experiments Ba. Compared with the blank group, the expression of Bax protein in the model group and the treatment group increased significantly (P0.01), and the expression of Bax protein in the treatment group increased (P0.05). (2) the expression of Bax in the treatment group and the treatment group was significantly lower than that in the model group (2). (3) the expression of Bax in the treatment group was higher than that in the treatment group (3) (P (P) (P) (P). (3) the expression of Bax in the treatment group was increased (P) (P). 0.05).4.3 Western blot P53 detection results: in the four groups, P53 had obvious bands, the measured gray value, (1) the expression of P53 protein in the model group and the treatment group increased significantly (P0.01), and the expression of P53 protein in the treatment group increased (P0.05), compared with the blank group (P0.05); (2) the expression of P53 protein in the treatment group 1 and the treatment group was significantly lower than that in the model group (P0.) 01) (3) compared with the treatment group 1, the expression of P53 protein in the treatment group was increased (P0.05).5. immunohistochemical detection results 5.1 Bcl-2 immunohistochemical detection results: four groups of experimental Bcl-2 protein expression, (1) compared with the blank group, the expression of Bcl-2 protein in the model group increased (P0.05), and the expression of Bcl-2 protein in the treatment group 1 and the treatment group was significantly increased (P0.0). 1) (1) (2) the expression of Bcl-2 protein in the treatment group was significantly higher than that in the model group (P0.01), and the expression of Bcl-2 protein in the treatment group increased (P0.05). (3) the expression of Bcl-2 protein expression increased (P0.05).5.2 Bax in the treatment group (1) compared with the treatment group (1): the Bax protein in the four groups was expressed, (1) compared with the blank, the model group and the treatment group 2 were compared. The expression of Bax protein in the treatment group increased significantly (P0.01), and the expression of Bax in the treatment group increased (P0.05). (2) the expression of Bax protein in the treatment group 1 and the treatment group was significantly lower (P0.01). (3) the expression of Bax protein expression in the treatment group (1) was increased (P0.05) in the treatment group (P0.05).5.3 P53 immunohistochemistry: (1) and (1) Compared with the blank group, the expression of P53 in the model group and the treatment group increased significantly (P0.01), and the expression of P53 in the treatment group increased (P0.05). (2) the expression of P53 in the treatment group 1 and the treatment group was significantly lower than that in the model group (P0.01). (3) the expression of P53 in the treatment group was higher than that in the treatment group (1) (P0.05). Conclusion: 1. electroacupuncture can improve the acute sciatic nerve injury in rats. The recovery of the motor function of the sciatic nerve, and the 2Hz group is better than the 100Hz group.2. electroacupuncture can regulate the apoptosis of the motor neuron in the anterior horn cell of the spinal cord after the acute sciatic nerve clamp injury and the expression of the related protein Bcl-2, Bax and p53. The mechanism may be related to the enhancement of the anti apoptotic factor Bcl-2, the decrease of the apoptosis stimulating factor Bax, and the expression of P53 in.3. electroacupuncture. Apoptosis of motor neurons in anterior horn of spinal cord and expression of related proteins Bcl-2, Bax and p53 after 2Hz were better than 100Hz.

【學(xué)位授予單位】：遼寧中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R245

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