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微囊化兔雪旺細胞移植后脊髓半橫斷損傷大鼠血清MBP動態(tài)變化及組織學觀察

發(fā)布時間:2018-03-26 14:42

  本文選題:微囊 切入點:雪旺細胞 出處:《南昌大學》2006年碩士論文


【摘要】:目的 觀察大鼠脊髓半橫斷損傷后植入微囊化兔雪旺細胞(Schwann Cells,SCs)對血清髓鞘堿性蛋白(Myelin Basic Protein,MBP)變化的影響及組織學變化,初步探討微囊化異種SCs移植治療脊髓損傷(Spinal Cord injury,SCI)的修復機制及對髓鞘再形成的影響。 方法 取健康成年家兔10只,體重2200±200g;Sprague-Dawley(SD)大鼠142只,體重220±20g,雌雄不拘,隨機分為4組:A組(單純損傷組,n=44);B組(SCs移植組,n=44);C組(微囊化SCs移植組,n=44);D組(正常對照組,n=10)。無菌條件下取成年家兔雙側坐骨神經干,利用反復差速貼附法體外培養(yǎng)SCs,制備SCs細胞懸液及微囊化SCs細胞懸液。建立大鼠T_(10)脊髓右半橫斷損傷模型,A、B、C組分別在損傷處移入吸附10μl的DMEM培養(yǎng)液、SCs懸液、微囊化SCs懸液的明膠海綿。D組為正常組。動物在灌注前,,心臟穿刺采血,用ELISA法檢測血清MBP。術后1d、3d、7d、14d、28d,各組每時相取8只(D組取2只)大鼠灌注固定,取損傷周圍處脊髓組織做石蠟切片,行HE、Loyez氏法髓鞘染色。另外,A、B、C組2w、8w時相點各2只大鼠,取損傷周圍處脊髓組織固定,Epon816包埋,醋酸鈾和檸檬酸鋁雙染,超薄切片電鏡觀察。 結果 各組損傷后1d即出現(xiàn)血清MBP升高,7d達到高峰,以后逐漸恢復。移植后1d、3d各組間血清MBP水平差異無顯著性(P>0.05)。移植后7d、14d、28d,與A、B組比較,C組血清MBP含量均較低(P<0.05),尤14d明顯(P<0.01);A組與B組比較,除術后7d血清MBP含量比較差異有顯著性(P<0.05)外,其他時相的比較差異無顯著性(P>0.05)。SCI后1d、3d各組損傷側脊髓神經元變性壞死,數(shù)量減少,部分神經元胞體皺縮、核固縮,細胞邊界模糊,髓鞘數(shù)量減少、結構松馳,脊髓組織內片狀出血,出血周圍可見神經膠質細胞和巨噬細胞浸潤,但B組和C組較少見。7d時壞死區(qū)有膠質細胞增生和白細胞浸
[Abstract]:Objective to observe the effect of microencapsulated Schwann cells of Schwann cells (Schwann Cellsberg SCS) on the serum myelin Basic protein (MBP) and its histological changes in rats after spinal cord hemitransection injury. To explore the repair mechanism of microencapsulated xenogeneic SCs transplantation for spinal cord injury (Spinal Cord injuryant) and its effect on myelin reformation. Methods Ten healthy adult rabbits, weighing 2200 鹵200g Sprague-Dawley SD rats, weighing 220 鹵20g, were selected. They were randomly divided into 4 groups: group A (simple injury group), group B (SCs transplantation group), group C (microencapsulated SCs transplantation group), group D (normal control group). Bilateral sciatic nerve trunk of adult rabbits was removed under aseptic condition. SCs cell suspensions and microencapsulated SCs cell suspensions were prepared by repeated differential attachment method in vitro. A rat model of right half transection of spinal cord was established. The microencapsulated SCs suspension of gelfoam group D was used as normal group. Blood samples were collected from heart puncture before perfusion, and serum MBP was detected by ELISA method. The rats were perfused and fixed with 8 rats in each group at 1 day, 3 days, 7 days, 14 days and 28 days after operation, and 8 rats in each group were taken from each group. The spinal cord tissue around the injury was taken as paraffin section and stained with Hefer Loyez's myelin sheath. In addition, two rats in group A and B (group C) were treated with Epon816 entrapped with fixed Epon816 at 8 weeks after injury. The specimens were stained with uranium acetate and aluminum citrate, and the ultrathin sections were observed by electron microscope. Results the serum MBP level increased 1 day after injury and reached the peak at 7 days after injury. There was no significant difference in serum MBP levels between the three groups on the 3rd day after transplantation (P > 0.05). The serum MBP levels in group C were lower than those in group A (P < 0.05), and those in group A were significantly lower than those in group B (P < 0.01) on the 7th day after transplantation (P < 0.01), and those in group B were significantly lower than those in group B (P < 0.01). Except for the significant difference in serum MBP content on the 7th day after operation (P < 0.05), there was no significant difference in other phases (P > P > 1 d after 0.05).SCI). The number of neurons in the injured side of the spinal cord was decreased, the number of neurons was reduced, some of the neurons were corrugated, nuclear pyknosis, and the cell boundaries were blurred. The number of myelin decreased, the structure relaxed, the lamellar hemorrhage of spinal cord tissue, the glial cells and macrophages infiltrated around the hemorrhage, but in group B and C there were glial cell proliferation and leucocyte leucocytosis in the necrotic area on the 7th day.
【學位授予單位】:南昌大學
【學位級別】:碩士
【學位授予年份】:2006
【分類號】:R651.2;R361

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