發(fā)布時間：2018-06-05 10:30

  本文選題：坐骨神經(jīng)損傷 + 活血通督湯　； 參考：《福建中醫(yī)藥大學》2017年碩士論文

【摘要】：目的通過觀察活血通督湯對坐骨神經(jīng)鉗夾傷大鼠脊髓運動神經(jīng)元的形態(tài)變化和相應節(jié)段脊髓組織iNOS、NGF、GDNF的表達情況,探討HXTDD保護脊髓神經(jīng)元的作用及機制。方法1、建立大鼠坐骨神經(jīng)鉗夾傷模型2月齡SD大鼠16只,隨機分為假手術(shù)組(8只)和模型組(8只)。模型組:大鼠麻醉后制成坐骨神經(jīng)鉗夾傷模型;假手術(shù)組只暴露坐骨神經(jīng),但不致傷神經(jīng)。兩組術(shù)后常規(guī)飼料喂養(yǎng)4w后取材,分別通過HE染色、尼氏染色,觀察光鏡下相應節(jié)段脊髓前角運動神經(jīng)元的形態(tài)變化和計數(shù)其神經(jīng)元存活率,并以免疫組化分析iNOS蛋白表達情況,鑒定造模是否成功。2、活血通督湯對坐骨神經(jīng)鉗夾傷模型大鼠脊髓的作用觀察2月齡SD大鼠48只,隨機分為假手術(shù)組16只,模型組16只,活血通督湯組16只。根據(jù)上述實驗造模操作,模型組和活血通督湯組建立大鼠坐骨神經(jīng)鉗夾傷模型,假手術(shù)組只暴露坐骨神經(jīng),但不致傷神經(jīng)。各組術(shù)后待動物蘇醒后立即開始給藥干預,活血通督湯組灌服活血通督湯,假手術(shù)組、模型組灌服等劑量0.9%氯化鈉溶液,每日2次,連續(xù)服用4w。給藥后每周記錄大鼠行為學變化情況。術(shù)后4周取材,相應節(jié)段脊髓組織行HE、尼氏染色法;免疫組化法檢測脊髓組織iNOS表達量;以WB、qPCR檢測各組大鼠脊髓iNOS、NGF、GDNF的蛋白及mRNA相對表達量。結(jié)果1、建立大鼠坐骨神經(jīng)鉗夾傷模型。2、大鼠行為學變化:術(shù)后每周進行行為學測定,活血通督湯組運動神經(jīng)功能恢復快于模型組,差異有統(tǒng)計學意義(P0.01,P0.05)。3、大鼠相應節(jié)段脊髓組織HE染色:假手術(shù)組:脊髓神經(jīng)細胞形態(tài)正常,細胞輪廓清晰,大而多角,核呈藍紫色,基質(zhì)著色均勻,有極性存在。模型組:脊髓神經(jīng)元結(jié)構(gòu)不清,形態(tài)多樣,基質(zhì)染色不均,細胞核縮小或不清,細胞周圍空泡形成明顯。活血通督湯組:其神經(jīng)細胞外貌較模型組規(guī)整,有散在的尼氏體,核仁較清楚,空泡樣改變極少,部分神經(jīng)元有水腫。4、大鼠相應節(jié)段脊髓前角運動神經(jīng)元存活率變化:術(shù)后4w,與假手術(shù)組相比,其余兩組的脊髓前角運動神經(jīng)元存活率顯著下降(P0.01),但活血通督湯組存活的運動神經(jīng)元較模型組多。5大鼠相應節(jié)段脊髓組織iNOS免疫組化法分析:模型組iNOS表達最高,且活血通督湯組明顯高于假手術(shù)組(P0.01)。6大鼠相應節(jié)段脊髓組織iNOS、NGF、GDNF的蛋白表達:藥物治療4周后,活血通督湯組GDNF、NGF的的蛋白表達水平較模型組明顯上升,iNOS表達較模型組有所下降,差異有統(tǒng)計學意義(P0.01,P0.05)。7大鼠相應節(jié)段脊髓組織iNOS、NGF、GDNFmRNA的表達:在受損脊髓組織中,iNOSmRNA的表達有所增強,模型組明顯高于活血通督湯組跟假手術(shù)組,差異有統(tǒng)計學意義(P0.05);脊髓組織GDNFmRNA和NGFmRNA表達,活血通督湯組、模型組均較假手術(shù)組高,而活血通督湯組也高于模型組表達(P0.05)。結(jié)論1、活血通督湯能改善坐骨神經(jīng)鉗夾傷模型大鼠后肢功能障礙;2、活血通督湯能有效改善坐骨神經(jīng)鉗夾傷模型大鼠脊髓前角運動神經(jīng)元的存活情況;3、活血通督湯能通過調(diào)控脊髓組織中iNOS、NGF、GDNF的表達,有效防治脊髓運動神經(jīng)元的損傷,為進一步深入研究提供基礎(chǔ)依據(jù)。
[Abstract]:Objective To observe the morphological changes of the spinal cord motoneurons and the expression of iNOS, NGF and GDNF in the corresponding segmental spinal cord tissue of the sciatic nerve clamp of rats with Huoxue Tong Du Decoction (Huoxue Tong Du Decoction), and to explore the effect and mechanism of HXTDD on the protection of spinal neurons. Method 1, 16 rats of the sciatic nerve clamp injury model of 2 month old SD rats were established and randomly divided into sham operation group (the group of 16 rats were randomly divided into sham operation group). 8 rats and model group (8). Model group: the rat model of sciatic nerve clamp injury was made after anesthesia. The sham operation group only exposed the sciatic nerve, but did not hurt the nerve. The two groups were fed after 4W and were fed with HE staining, Nissl staining, and observed the morphological changes of the motor neurons in the anterior horn of the spinal cord under the light microscope and the count of the nerve. The survival rate of the iNOS protein was analyzed by immunohistochemistry. The effect of.2 was identified and the effect of Huoxue Tong Du Decoction on the spinal cord of the sciatic nerve clamp was observed in 2 month old SD rats, and 16 rats were randomly divided into a sham operation group, 16 in the model group and 16 in the Huoxue Tong Du Decoction group. The rat model of the sciatic nerve clamp injury was established in the Tong Du Decoction group. The sham operation group only exposed the sciatic nerve, but did not hurt the nerve. After the animals were awakened, the rats were given the medicine intervention immediately after the animals were awakened. The group of Huoxue Tong Du Decoction group was filled with Huoxue Tong Du decoction, sham operation group, and the model group was given the dosage of 0.9% Sodium Chloride Solution, 2 times a day, every week after the continuous administration of 4w. administration. The changes in the behavior of the rats were recorded. 4 weeks after the operation, HE, Nissl staining was used in the corresponding segments of the spinal cord, and the expression of iNOS in the spinal cord tissue was detected by immunohistochemistry; the iNOS, NGF, GDNF protein and mRNA relative expression of iNOS, NGF, GDNF in the spinal cord of rats in each group were detected by WB and qPCR. Results 1, the rat sciatic nerve clamp injury model.2 and the behavior changes of rats were established. After a weekly behavioral test, the motor nerve function of the Huoxue Tong Du Decoction group was recovered faster than the model group. The difference was statistically significant (P0.01, P0.05).3, and the corresponding spinal cord tissue of the rat was HE staining: the sham operation group: the spinal nerve cell morphology was normal, the cell outline was clear, the large and multi angle, the nucleus was blue purple, the matrix coloring was even and polar existed. There was polarity existence. Type group: spinal cord neuron structure is not clear, morphological diversity, uneven matrix staining, cell nuclei narrowing or unclear, cell surrounding vacuoles formation obviously. Huoxue Tong Du group: the nerve cell appearance is more regular than the model group, there are scattered Nissl body, the nucleolus is clearer, the vacuoles change very few, some neurons have edema.4, the corresponding segment of the spinal cord anterior horn of the rat The survival rate of motor neuron: 4W after operation, compared with the sham group, the survival rate of the motor neuron in the other two groups decreased significantly (P0.01), but the surviving motoneurons in the Huoxue Tong Du Decoction group were compared with that of the model group in the corresponding segment spinal cord tissue with iNOS immunohistochemical method: the iNOS expression of the model group was the highest and the Huoxue Tong Du Decoction group was in the model group. The protein expression of iNOS, NGF, GDNF in the corresponding segment of the spinal cord of the sham group (P0.01).6 rats was significantly higher than that of the model group after 4 weeks of drug treatment, and the expression level of GDNF in the Huoxue Tong Du Decoction group was significantly higher than that in the model group, and the expression of iNOS was lower than that in the model group, and the difference was statistically significant (P0.01, P0.05).7 rat spinal cord tissue iNOS. The expression of GDNFmRNA: in the damaged spinal cord tissue, the expression of iNOSmRNA was enhanced. The model group was significantly higher than the Huoxue Tong Du Decoction group and the sham group (P0.05). The expression of GDNFmRNA and NGFmRNA in the spinal cord, the Huoxue Tong Du Decoction group, the model group were higher than the sham operation group, and the Huoxue Tong Du Decoction group was also higher than the model group (P0.05 Conclusion 1. 1, Huoxue Tong Du decoction can improve the hindlimb dysfunction of the sciatic nerve clamp injury model rats. 2, Huoxue Tong Du decoction can effectively improve the survival of the motor neurons of the anterior horn of the spinal cord of the sciatic nerve clamp injury model rats. 3, Huoxue Tong Du decoction can effectively control the spinal motor neurons by regulating the expression of iNOS, NGF, and GDNF in the spinal cord tissue. The damage will provide a basis for further study.
【學位授予單位】：福建中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R285.5;R-332

【參考文獻】

相關(guān)期刊論文 前10條

1 吳楊鵬;范筱;張俐;;活血通督湯對大鼠急性脊髓損傷后神經(jīng)生長因子和膠質(zhì)纖維酸蛋白質(zhì)表達的影響[J];中華中醫(yī)藥雜志;2017年02期

2 邵航;陳芬芳;張俐;;活血通督湯對坐骨神經(jīng)損傷后脊髓損傷性反應的作用機制[J];中國中醫(yī)骨傷科雜志;2016年06期

3 何新澤;王維;呼鐵民;馬建軍;于昌玉;高云峰;程興龍;王培;;周圍神經(jīng)損傷的修復:理論研究與技術(shù)應用[J];中國組織工程研究;2016年07期

4 魯夢倩;于天源;姚斌彬;陳國勇;潘t，

本文編號：1981668