本文選題：脊髓損傷　切入點(diǎn)：神經(jīng)干細(xì)胞　出處：《中國(guó)組織工程研究》2017年25期 　論文類(lèi)型：期刊論文

【摘要】：背景:有研究顯示舒芬太尼聯(lián)合神經(jīng)干細(xì)胞移植聯(lián)合治療可以提高新生的神經(jīng)纖維數(shù)量。目的:觀察舒芬太尼干預(yù)對(duì)神經(jīng)干細(xì)胞移植治療脊髓損傷大鼠后肢功能恢復(fù)的影響。方法:(1)將100只成年雌性SD大鼠隨機(jī)分為5組:正常對(duì)照組、模型組、舒芬太尼組、神經(jīng)干細(xì)胞移植組和舒芬太尼+神經(jīng)干細(xì)胞移植聯(lián)合治療組,每組20只。正常對(duì)照組大鼠不實(shí)施任何干預(yù),其他4組參照改良的Allen’s重物打擊法建立大鼠脊髓損傷模型;(2)造模6 h后,聯(lián)合治療組大鼠蛛網(wǎng)膜下腔注射10μL細(xì)胞濃度為1×10~(10) L~(-1)的神經(jīng)干細(xì)胞懸液,腹腔注射100μL舒芬太尼(150μg/kg);神經(jīng)干細(xì)胞移植組大鼠蛛網(wǎng)膜下腔注射10μL細(xì)胞濃度為1×10~(10) L~(-1)的神經(jīng)干細(xì)胞懸液,腹腔注射100μL生理鹽水;舒芬太尼組蛛網(wǎng)膜下腔注射10μL的神經(jīng)干細(xì)胞培養(yǎng)液,腹腔注射100μL舒芬太尼(150μg/kg);模型組大鼠蛛網(wǎng)膜下腔和腹腔各注射10μL神經(jīng)干細(xì)胞培養(yǎng)液和100μL生理鹽水;(3)造模后72 h,RT-PCR檢測(cè)脊髓損傷區(qū)周?chē)鶤QP4、MMP9 mRNA的表達(dá),TUNEL法檢測(cè)細(xì)胞凋亡情況;(4)造模后第1,3天和第1,2,3,4周通過(guò)BBB評(píng)分、斜板實(shí)驗(yàn)進(jìn)行運(yùn)動(dòng)功能評(píng)定;(5)造模后4周取材行蘇木精-伊紅染色,熒光顯微鏡觀測(cè)CM-Dil標(biāo)記的神經(jīng)干細(xì)胞存活情況,熒光金逆行追蹤觀察脊髓神經(jīng)纖維的再生與分布情況。結(jié)果與結(jié)論:(1)造模后72 h,與模型組、舒芬太尼組、神經(jīng)干細(xì)胞移植組比較,聯(lián)合治療組脊髓損傷區(qū)AQP4、MMP9 mRNA表達(dá)和細(xì)胞凋亡率顯著降低(P0.05);(2)造模2周后,聯(lián)合治療組運(yùn)動(dòng)功能評(píng)分優(yōu)于舒芬太尼組和神經(jīng)干細(xì)胞移植組(P0.05),舒芬太尼組和神經(jīng)干細(xì)胞移植組優(yōu)于模型組(P0.05);(3)造模后4周,模型組可見(jiàn)脊髓空洞形成,舒芬太尼組和神經(jīng)干細(xì)胞移植組損傷區(qū)脊髓空洞較小,聯(lián)合治療組脊髓空洞幾乎消失;(4)聯(lián)合治療組CM-Dil陽(yáng)性細(xì)胞最多,神經(jīng)干細(xì)胞移植組較多,舒芬太尼組和模型組未見(jiàn)陽(yáng)性細(xì)胞;(5)聯(lián)合治療組熒光金陽(yáng)性神經(jīng)纖維數(shù)最多,舒芬太尼組和神經(jīng)干細(xì)胞移植組次之,模型組最少;(6)結(jié)果表明,舒芬太尼通過(guò)促進(jìn)移植神經(jīng)干細(xì)胞的增殖,減少大鼠脊髓損傷區(qū)AQP4、MMP9表達(dá)和局部神經(jīng)細(xì)胞凋亡,進(jìn)而改善后肢運(yùn)動(dòng)功能。
[Abstract]:Background: studies have shown that sufentanil combined with neural stem cell transplantation can increase the number of newborn nerve fibers. Objective: to observe the effects of sufentanil on the function of hindlimb of spinal cord injury rats treated by neural stem cell transplantation. Methods 100 adult female SD rats were randomly divided into 5 groups: normal control group. Model group, sufentanil group, neural stem cell transplantation group and sufentanil neural stem cell transplantation combined treatment group, 20 rats in each group. In the other four groups, the spinal cord injury model of rats was established by using the modified Allen's weight attack method for 6 h. The rats in the combined treatment group were injected into subarachnoid space with 10 渭 L cell concentration of 1 脳 10 ~ (10) L ~ (-1) L ~ (-1) neural stem cell suspension. 100 渭 L sufentanil 150 渭 g 路kg ~ (-1) was injected intraperitoneally into 100 渭 L sufentanil 150 渭 g 路kg ~ (-1), 10 渭 L neural stem cell suspension (10 渭 L cell concentration was 1 脳 10 ~ (10)) L ~ (-1) in the neural stem cell transplantation group, 100 渭 L normal saline was injected intraperitoneally, and 10 渭 L neural stem cell culture medium was injected into the subarachnoid space of sufentanil group. 100 渭 L sufentanil 150 渭 g / kg; subarachnoid and intraperitoneal injection of 10 渭 L neural stem cell culture medium and 100 渭 L normal saline in model group) 72 h later, RT-PCR was used to detect the expression of AQP4MMP9 mRNA around spinal cord injury area and Tunel method was used to detect apoptosis. The BBB score was obtained on the 1st 3rd day and 4th week of the 1st day and the 2nd week after the model was made. Four weeks after the model was made, the rats were stained with hematoxylin and eosin, and the survival of neural stem cells labeled with CM-Dil was observed by fluorescence microscope. The regeneration and distribution of nerve fibers in spinal cord were observed by fluorescence gold retrograde tracing. Results and conclusion the results were compared with those of model group, sufentanil group and neural stem cell transplantation group at 72 h after the establishment of the model group, sufentanil group and neural stem cell transplantation group. In the combined treatment group, the expression of MMP9 mRNA and the apoptotic rate of AQP4mMP9 in spinal cord injury area were significantly decreased 2 weeks after the establishment of the model. The motor function score in the combined treatment group was better than that in the sufentanil group and the neural stem cell transplantation group (P 0.05), and in the sufentanil group and the neural stem cell transplantation group, the spinal cord syringomyelia was found in the model group 4 weeks after the establishment of the model. In sufentanil group and neural stem cell transplantation group, syringomyelia in the injured area was smaller, and syringomyelia almost disappeared in the combined treatment group. The number of CM-Dil positive cells in the combined treatment group was more than that in the neural stem cell transplantation group. No positive cells were found in Sufentanil group and model group. The number of fluorescent gold positive nerve fibers was the highest in combined treatment group, followed by sufentanil group and neural stem cell transplantation group. Sufentanil could improve the motor function of hindlimb by promoting the proliferation of transplanted neural stem cells, reducing the expression of AQP4MMP9 and local neuronal apoptosis in spinal cord injury area of rats.
【作者單位】： 濱州醫(yī)學(xué)院附屬醫(yī)院麻醉科;濱州醫(yī)學(xué)院附屬醫(yī)院藥劑科;
【分類(lèi)號(hào)】：R651.2

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