賴氨酰氧化酶在大鼠脊髓慢性受壓中的作用
發(fā)布時(shí)間：2019-07-08 09:43
【摘要】：目的通過(guò)漸進(jìn)性螺釘植入法成功復(fù)制出慢性脊髓損傷的大鼠模型的基礎(chǔ)上,比較賴氨酰氧化酶促進(jìn)組、賴氨酰氧化酶抑制組、單純慢性損傷壓迫對(duì)照組各組間的不同,探討賴氨酰氧化酶在大鼠脊髓慢性受壓中的作用。觀察不同組間皮層體感誘發(fā)電位、慢性脊髓損傷后組織結(jié)構(gòu)變異的關(guān)系探討賴氨酰氧化酶對(duì)于慢性脊髓壓迫的治療意義。方法健康SD雌性大鼠隨機(jī)分為四組:空白對(duì)照組(A組,n=15);單純慢性損傷壓迫對(duì)照組(B組,n=15);壓迫+BAPN LOX抑制組(C組,n=15);壓迫+氯化鈷LOX促進(jìn)組(D組,n=15),建立慢性脊髓壓迫模型后。B組,致傷后不做任何治療處理,C組BAPN 1g/KG(腹腔注射),D組Co Cl2 30mg/L(給水)。飼養(yǎng)至8周以后,進(jìn)行BBB評(píng)分、Tarlov運(yùn)動(dòng)評(píng)分及CSEP檢測(cè)后獲取大鼠脊髓部分組織進(jìn)行RT-PCR檢測(cè)賴氨酰氧化酶表達(dá)含量,部分組織制成石蠟包塊兒進(jìn)行HE及免疫組化染色。結(jié)果經(jīng)脊髓組織RT-PCR檢測(cè)結(jié)果顯示與空白對(duì)照組A組相比各壓迫組大鼠的脊髓內(nèi)賴氨酰氧化酶的表達(dá)含量均有不同程度的升高。其中LOX促進(jìn)組D組升高最為明顯,這說(shuō)明氯化鈷成功的促進(jìn)了賴氨酰氧化酶的表達(dá)。而LOX抑制組C組升高的程度最低,說(shuō)明β氨基丙晴按照預(yù)期的設(shè)想抑制了賴氨酰氧化酶的表達(dá)。CSEP實(shí)驗(yàn)結(jié)果顯示隨時(shí)間變化除對(duì)照組A潛伏期未見(jiàn)變化外,其余各組潛伏期均出現(xiàn)不同程度的延長(zhǎng)。測(cè)得各壓迫組間結(jié)果在6周后出現(xiàn)差異,在8周差異明顯。第8周時(shí)LOX促進(jìn)組潛伏期為(18.06±0.67)ms,單純慢性壓迫陽(yáng)性對(duì)照組為(19.76±0.82)ms,LOX抑制組為(22.16±0.74)ms。HE染色結(jié)果顯示LOX抑制組D組與LOX促進(jìn)組C組相比較白質(zhì)中可見(jiàn)腫脹的軸突,軸突周圍的空泡也較多,組織水腫較為嚴(yán)重、結(jié)構(gòu)更為紊亂,細(xì)胞成分明顯減少;灰質(zhì)中可見(jiàn)神經(jīng)元細(xì)胞皺縮更為明顯,細(xì)胞核固縮,尼氏體更少甚至消失。利用免疫組化技術(shù)觀察Bcl-2/Bax的表達(dá)情況從而檢測(cè)脊髓神經(jīng)細(xì)胞的凋亡情況。Bcl-2在各組中的表達(dá)水平為:LOX空白對(duì)照組A組的光密度值為(0.48±0.01),單純慢性損傷壓迫對(duì)照組B組的光密度值為(0.36±0.02),LOX抑制組C組的光密度值(0.25±0.02),LOX促進(jìn)組D組的光密度值(0.31±0.01)。各組比較差異明顯有統(tǒng)計(jì)學(xué)意義(P0.05)。Bax在空白對(duì)照組A組的表達(dá)最低,其余三組表達(dá)均升高,其中C組升高最為明顯。D組較C組有所降低且差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論賴氨酰氧化酶可能在脊髓的慢性受壓過(guò)程中起著重要的作用。
[Abstract]:Objective To study the model of rats with chronic spinal cord injury by means of progressive screw implantation. Objective To study the role of aminopeptidase in the chronic compression of spinal cord in rats. To observe the relationship between somatosensory evoked potential and chronic spinal cord injury in different groups, and to explore the significance of the method for the treatment of chronic spinal cord compression in the patients with chronic spinal cord injury. Methods Healthy SD female rats were randomly divided into four groups: blank control group (group A, n = 15), simple chronic injury compression control group (group B, n = 15), compression + BAPN LOX inhibition group (group C, n = 15), compression + cobalt chloride-type LOX promotion group (group D, n = 15), and establishment of chronic spinal cord compression model. Group B, no treatment after injury, group C BAPN 1g/ KG (intraperitoneal injection), and D group Co Cl2 30 mg/ L (water supply). After feeding for 8 weeks, BBB score, Tarlov's motion score and CSEP detection were performed to detect the expression level of the insulin-aminidase oxidase in the spinal cord of the rat, and the tissue was made into a paraffin-containing block for HE and immunohistochemical staining. Results The results of RT-PCR in the spinal cord showed that the level of the expression of the myeloperoxidase in the spinal cord of each compression group was higher than that of the control group A. In which LOX promoted the most significant increase in group D, suggesting that cobalt chloride successfully promoted the expression of the amine oxidase. While the level of the elevation of the group C in the LOX-inhibiting group was the lowest, and the inhibition of the expression of the aminopeptidase was inhibited according to the expected assumption. The results of CSEP showed that there was no change in the latency of the control group A, and the latency of the remaining groups was extended to a different extent. The difference in the results of each compression group after 6 weeks was measured, and the difference was evident at 8 weeks. At week 8, the latency of the LOX-promoting group was (18.06-0.67) ms, the simple chronic compression-positive control group was (19.76-0.82) ms, and the LOX-inhibiting group (22.16-0.74) ms. HE staining showed that the LOX-inhibiting group D and the LOX-promoting group C group compared the axons of the visible swelling in the white matter, The vacuoles around the axons are also more, the edema of the tissue is more serious, the structure is more disordered, the cell components are obviously reduced, the cells of the visible neurons in the gray matter are more obvious, the cells are fixed, and the ntroostite is less and even disappears. The expression of Bcl-2/ Bax was observed by immunohistochemistry in order to detect the apoptosis of spinal cord. The expression level of Bcl-2 in the control group was (0.48-0.01), the optical density value of group B was (0.36-0.02), and the optical density (0.25-0.02) in group C was inhibited in the control group. The optical density values of the group D were promoted by LOX (0.31-0.01). The expression of Bax in group A of blank control group was the lowest, and the expression of the other three groups increased, among which group C was the most obvious. There was a significant difference between group D and group C (P0.05). Conclusion The myeloperoxidase may play an important role in the chronic compression of the spinal cord.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2
本文編號(hào)：2511500
[Abstract]:Objective To study the model of rats with chronic spinal cord injury by means of progressive screw implantation. Objective To study the role of aminopeptidase in the chronic compression of spinal cord in rats. To observe the relationship between somatosensory evoked potential and chronic spinal cord injury in different groups, and to explore the significance of the method for the treatment of chronic spinal cord compression in the patients with chronic spinal cord injury. Methods Healthy SD female rats were randomly divided into four groups: blank control group (group A, n = 15), simple chronic injury compression control group (group B, n = 15), compression + BAPN LOX inhibition group (group C, n = 15), compression + cobalt chloride-type LOX promotion group (group D, n = 15), and establishment of chronic spinal cord compression model. Group B, no treatment after injury, group C BAPN 1g/ KG (intraperitoneal injection), and D group Co Cl2 30 mg/ L (water supply). After feeding for 8 weeks, BBB score, Tarlov's motion score and CSEP detection were performed to detect the expression level of the insulin-aminidase oxidase in the spinal cord of the rat, and the tissue was made into a paraffin-containing block for HE and immunohistochemical staining. Results The results of RT-PCR in the spinal cord showed that the level of the expression of the myeloperoxidase in the spinal cord of each compression group was higher than that of the control group A. In which LOX promoted the most significant increase in group D, suggesting that cobalt chloride successfully promoted the expression of the amine oxidase. While the level of the elevation of the group C in the LOX-inhibiting group was the lowest, and the inhibition of the expression of the aminopeptidase was inhibited according to the expected assumption. The results of CSEP showed that there was no change in the latency of the control group A, and the latency of the remaining groups was extended to a different extent. The difference in the results of each compression group after 6 weeks was measured, and the difference was evident at 8 weeks. At week 8, the latency of the LOX-promoting group was (18.06-0.67) ms, the simple chronic compression-positive control group was (19.76-0.82) ms, and the LOX-inhibiting group (22.16-0.74) ms. HE staining showed that the LOX-inhibiting group D and the LOX-promoting group C group compared the axons of the visible swelling in the white matter, The vacuoles around the axons are also more, the edema of the tissue is more serious, the structure is more disordered, the cell components are obviously reduced, the cells of the visible neurons in the gray matter are more obvious, the cells are fixed, and the ntroostite is less and even disappears. The expression of Bcl-2/ Bax was observed by immunohistochemistry in order to detect the apoptosis of spinal cord. The expression level of Bcl-2 in the control group was (0.48-0.01), the optical density value of group B was (0.36-0.02), and the optical density (0.25-0.02) in group C was inhibited in the control group. The optical density values of the group D were promoted by LOX (0.31-0.01). The expression of Bax in group A of blank control group was the lowest, and the expression of the other three groups increased, among which group C was the most obvious. There was a significant difference between group D and group C (P0.05). Conclusion The myeloperoxidase may play an important role in the chronic compression of the spinal cord.
【學(xué)位授予單位】：華北理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R651.2
【共引文獻(xiàn)】
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，本文編號(hào)：2511500
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