本文選題：槲皮素 + 脊髓損傷��； 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:通過(guò)使用黃酮類(lèi)化合物槲皮素治療大鼠急性脊髓損傷(spinal cord injury,SCI),驗(yàn)證其治療效果,并探討其可能的作用機(jī)制。方法:取128只成年健康雌性SD大鼠,隨機(jī)等分為對(duì)照組、干預(yù)組、治療組和假手術(shù)組。對(duì)照組、干預(yù)組及治療組大鼠使用自制脊髓損傷打擊器建模:常規(guī)麻醉并消毒,摘除T9-T11椎板后暴露相應(yīng)脊髓節(jié)段,采用10g×25mm致傷勢(shì)能對(duì)脊髓進(jìn)行垂直打擊建模。治療組及干預(yù)組大鼠于術(shù)后1h給予腹腔注射槲皮素(25μmol/kg,12h/次),持續(xù)7日;干預(yù)組大鼠在注射槲皮素后立即在對(duì)側(cè)腹腔注射p38MAPK特異性激活劑anisomycin;假手術(shù)組大鼠僅行椎板切除術(shù)。對(duì)照組及假手術(shù)組大鼠術(shù)后予以與治療組等量、等頻率的生理鹽水腹腔注射。大鼠建模成功后按摩膀胱排尿3次/日,直到恢復(fù)反射性膀胱;術(shù)后7日內(nèi)所有大鼠行氨芐青霉素腹腔注射預(yù)防感染。術(shù)后1d、3d、7d、14d、21d及28d對(duì)各組大鼠進(jìn)行BBB評(píng)分。術(shù)后3d、7d、14d及28d時(shí)每組隨機(jī)取8只大鼠麻醉并行心臟灌注,灌注成功后切取長(zhǎng)約1cm的脊髓組織,縱切為均勻的左右兩半,取其中一半保存于-80℃冰箱,以備行Western blot蛋白檢測(cè),另一半組織在4%多聚甲醛中浸泡24h用于免疫組化或HE染色。術(shù)后28d對(duì)各組大鼠脊髓組織進(jìn)行HE染色并在顯微鏡下觀察。術(shù)后3d、7d、14d及28d對(duì)各組大鼠脊髓組織的膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein,GFAP)和神經(jīng)絲蛋白-200(neurofilament protein-200,NF-200)進(jìn)行免疫組化染色,顯微鏡下觀察,統(tǒng)計(jì)分析各組的陽(yáng)性細(xì)胞數(shù)目。術(shù)后3d、7d、14d及28d對(duì)各組大鼠脊髓組織的p38MAPK、磷酸化p38MAPK(phosphorylation p38MAPK,p-p38MAPK)蛋白的表達(dá)情況進(jìn)行Western blot檢測(cè),術(shù)后28d對(duì)各組大鼠脊髓組織的GFAP和NF-200蛋白進(jìn)行Western blot檢測(cè)。結(jié)果:術(shù)后7d、14d、21d及28d,治療組大鼠BBB評(píng)分顯著高于對(duì)照組與干預(yù)組(P0.05)。術(shù)后3d和7d各脊髓損傷組的p38MAPK磷酸化水平明顯高于假手術(shù)組(P0.05),治療組的磷酸化水平在術(shù)后3d、7d及14 d時(shí)明顯低于對(duì)照組與干預(yù)組(P0.05)。各脊髓損傷組在術(shù)后各時(shí)間點(diǎn)NF-200和GFAP陽(yáng)性細(xì)胞數(shù)均高于假手術(shù)組(P0.05),治療組的NF-200陽(yáng)性細(xì)胞數(shù)較對(duì)照組和干預(yù)組顯著增高(P0.05);術(shù)后7d、14d和28d,治療的GFAP陽(yáng)性細(xì)胞數(shù)明顯低于對(duì)照組與干預(yù)組(P0.05)。結(jié)論:槲皮素通過(guò)抑制急性脊髓損傷后p38MAPK信號(hào)通路的激活,促進(jìn)了 NF-200的表達(dá)及神經(jīng)元軸突再生,抑制了 GFAP表達(dá)及星形膠質(zhì)細(xì)胞的增生、阻礙了膠質(zhì)瘢痕的形成,修復(fù)受損脊髓的神經(jīng)傳導(dǎo)作用,從而促進(jìn)急性脊髓損傷后運(yùn)動(dòng)功能的恢復(fù)。
[Abstract]:Aim: to investigate the therapeutic effect of quercetin on acute spinal cord injury (spinal cord injura) in rats and its possible mechanism. Methods: 128 adult female SD rats were randomly divided into control group, intervention group, treatment group and sham operation group. Control group, intervention group and treatment group used self-made spinal cord injury batter model: routine anesthesia and disinfection, after removing T9-T11 vertebral lamina, the corresponding spinal cord segment was exposed. The injury induced by 10 g 脳 25mm could be used to model the spinal cord vertical strike. Rats in the treatment group and the intervention group were intraperitoneally injected with quercetin (25 渭 mol / kg, 12 h / time) for 7 days, and the rats in the intervention group were injected with p38 MAPK specific activator anisomycin in the contralateral abdominal cavity immediately after injection of quercetin, while rats in the sham operation group were treated with laminectomy only. Rats in the control group and sham operation group were given intraperitoneal injection of saline with the same amount and frequency after operation as the treatment group. After successful modeling, the bladder urination was massaged 3 times a day until the reflex bladder was restored, and all rats received ampicillin intraperitoneal injection to prevent infection within 7 days after operation. BBB scores were evaluated on the 1st day, 3rd day, 7th day, 14 d, 21 d and 28 d after operation. 8 rats in each group were anesthetized and perfused on the 14th and 28th days after the operation. The spinal cord tissue about 1cm was cut out and divided into two parts, half of which was stored in the refrigerator at -80 鈩，

本文編號(hào)：2115583