本文選題：基膜　切入點(diǎn)：慢性脊髓壓迫　出處：《中國脊柱脊髓雜志》2016年02期

【摘要】：目的 :觀察慢性頸脊髓壓迫癥大鼠模型髓內(nèi)基膜(basement membrane,BM)、基膜與星形細(xì)胞接觸面(basement membrane-astrocyte contacts,BM-AC)的超微結(jié)構(gòu)變化,并探討其與基質(zhì)金屬蛋白酶-9(matrix metalloproteinase-9,MMP-9)表達(dá)的相關(guān)性。方法 :72只雄性SD大鼠隨機(jī)分為對照組(n=36)和實(shí)驗(yàn)組(n=36),對照組僅切除C5左側(cè)椎板;實(shí)驗(yàn)組在切除C5左側(cè)椎板后將吸水可膨脹聚氨酯薄板置入C6水平左側(cè)椎板下硬膜外,建立慢性頸脊髓壓迫模型,應(yīng)用BBB(Basso Beattie Bresnahan)評分評價大鼠脊髓神經(jīng)功能,并分別于造模后1d、14d、21d、28d、42d、70d取C5~C6段脊髓組織制備標(biāo)本,用HE染色觀察脊髓形態(tài)學(xué)變化、用MMP-9免疫組化染色檢測脊髓MMP-9表達(dá)量,用透射電鏡觀察脊髓BM及BM-AC的變化。結(jié)果:對照組各時間點(diǎn)間BBB評分和實(shí)驗(yàn)組造模后1d的BBB評分無顯著性差異,實(shí)驗(yàn)組造模后14d~70d的5個時間點(diǎn)BBB評分均顯著性低于同時間點(diǎn)對照組(P0.05)。HE染色顯示對照組各時間點(diǎn)及實(shí)驗(yàn)組造模后1d的脊髓未見受壓,脊髓形態(tài)結(jié)構(gòu)正常;實(shí)驗(yàn)組造模后1d可見脊髓白質(zhì)區(qū)輕度水腫;造模后14d脊髓受壓變形,灰質(zhì)區(qū)血管增生,灰質(zhì)、白質(zhì)水腫,神經(jīng)元細(xì)胞核碎裂;造模后21d和28d損傷逐漸加重;造模后42d脊髓水腫減輕,髓內(nèi)空泡化,前角大運(yùn)動神經(jīng)元數(shù)目減少、胞漿稀少、胞核萎縮,突觸減少,神經(jīng)纖維稀疏,髓鞘層變薄;造模后70d仍見白質(zhì)區(qū)水腫、神經(jīng)元細(xì)胞核碎裂,灶性膠質(zhì)細(xì)胞增生等退行性變,神經(jīng)元數(shù)目增多。MMP-9免疫組化顯示對照組各時間點(diǎn)及實(shí)驗(yàn)組造模后1d、70d脊髓MMP-9均呈弱表達(dá),實(shí)驗(yàn)組造模后14d呈較強(qiáng)表達(dá),21d呈強(qiáng)表達(dá),28d呈較強(qiáng)表達(dá),42d呈中度表達(dá)。對照組各時間點(diǎn)及實(shí)驗(yàn)組壓迫后1d的BM電子密度及BM-AC均正常,實(shí)驗(yàn)組造模后14d~28d BM電子密度、BM-AC比率與對照組比較顯著性降低(P0.05);實(shí)驗(yàn)組造模后42d、70d兩者較前升高(P0.05),但仍顯著低于對照組水平(P0.05)。MMP-9表達(dá)與BM電子密度及BM-AC變化呈負(fù)相關(guān),相關(guān)系數(shù)分別為-0.892(P0.001)和-0.664(P0.001)。結(jié)論:慢性頸脊髓壓迫性損傷后早期髓內(nèi)BM降解、BMAC分離,后期部分修復(fù)。MMP-9可能通過降解BM及BM-AC影響脊髓壓迫后血脊髓屏障的完整性。
[Abstract]:Objective: to observe the ultrastructural changes of the intramedullary basement membrane membrane-astrocyte (BM-AC) in rats with chronic cervical spinal cord compression. Methods Seventy-two male Sprague-Dawley rats were randomly divided into control group (n = 36) and experimental group (n = 36). In the experimental group, the water-absorbing expandable polyurethane thin plate was placed into the subdural space of the C6 level, and the chronic cervical spinal cord compression model was established. The spinal cord nerve function was evaluated by BBB(Basso Beattie Bresnahan score. The spinal cord tissue of the C5~C6 segment was taken from the spinal cord on the 1st day, 14d, 21d, 28d and 42d, respectively. The morphological changes of the spinal cord were observed by HE staining, and the expression of MMP-9 in the spinal cord was detected by MMP-9 immunohistochemical staining. The changes of BM and BM-AC in spinal cord were observed by transmission electron microscope. Results: there was no significant difference in BBB score between the control group and the BBB score of the experimental group at 1 day after modeling. The BBB scores of 14d~70d at 5 time points in the experimental group were significantly lower than those in the control group at the same time point (P 0.05). He staining showed that the spinal cord of the control group and the experimental group had no compression at each time point and 1 day after the model making, and the morphology and structure of the spinal cord were normal. Mild edema in the white matter area of the spinal cord was observed in the experimental group on the 1st day after modeling, compression deformation of the spinal cord was observed on the 14th day after modeling, vascular proliferation, gray matter, white matter edema and nuclear fragmentation of the neurons in the gray matter area, and the injury increased gradually on the 21st and 28th days after the establishment of the model. The edema of spinal cord was alleviated 42 days later, the intramedullary vacuolation, the number of major motor neurons in the anterior horn decreased, the cytoplasm was scarce, the nucleus atrophy, the synapse decreased, the nerve fibers were sparse, the myelin sheath became thinner, and the edema of the white matter area was still observed 70 days after the model. The neuronal nucleus fragmentation, focal glial cell proliferation and other degenerative changes. The number of neurons increased. MMP-9 immunohistochemical staining showed that the spinal cord MMP-9 was weakly expressed at each time point in the control group and on the 70th day after modeling in the experimental group. The experimental group showed strong expression on the 14th day and the middle expression on the 28th day, and the BM electron density and BM-AC of the control group and the experimental group were normal at each time point and 1 day after compression. The BM-AC ratio of 14d~28d BM in the experimental group was significantly lower than that in the control group (P 0.05A), and the expression of BM-AC in the experimental group was significantly lower than that in the control group (P 0.05), but the expression of MMP-9 was negatively correlated with the changes of BM electron density and BM-AC. The correlation coefficients were -0.892g / P0.001) and -0.664m / P0.001respectively. Conclusion: BMAC separation of BM degradation in early stage after chronic cervical spinal cord compression injury, partial repair of MMP-9 may affect the integrity of blood-spinal cord barrier after spinal cord compression by degradation of BM and BM-AC.
【作者單位】： 中山大學(xué)附屬第一醫(yī)院東院脊柱外科;中山大學(xué)附屬第一醫(yī)院東院神經(jīng)外科;
【基金】：國家自然科學(xué)基金項(xiàng)目(編號:81450020) 廣東省自然科學(xué)基金項(xiàng)目(編號:S2013010015778)
【分類號】：R744

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