【摘要】 目的:观察大鼠脊髓损伤后转化生长因子β1mRNA表达的变化,及甲基强的松龙对其影响。方法:实验于2003-06/2004-03在上海市药物工业研究院与解放军第二军医大学长海医院病理实验室完成。选择雄性SD大鼠45只,随机分为空白对照组5只、损伤对照组(6,24h,3,7d)、甲基强的松龙组(6,24h,3,7d),每时间点5只。制作脊髓损伤模型后,损伤对照组及空白对照组给予等渗盐水,甲基强的松龙组给予甲基强的松龙(30mg/kg),分别于伤后6,24h,3,7d切取损伤及临近部位脊髓组织,运用原位杂交技术,二氨基联苯胺显色,苏木精轻度复染。采用光学显微镜进行观察,高倍镜下(200倍),随机选择5个高表达视野,记数每个高倍镜视野下阳性细胞的百分比(阳性细胞数/细胞总数)。细胞胞浆内呈棕黄色为转化生长因子β1mRNA。观察甲基强的松龙对大鼠脊髓损伤转化生长因子β1mRNA表达的影响。结果:纳入动物45只,均进入结果分析。①脊髓损伤后甲基强的松龙组转化生长因子β1mRNA的表达阳性细胞数与损伤对照组相比明显降低眼损伤对照组伤后6,24h,3,7d分别为(8.91±0.82)%,(28.27±1.10)%,(42.09±0.93)%,(45.24±0.85)%,甲基强的松龙组伤后6,24h,3,7d分别为(7.93±0.40)%,(20.36±1.10)%,(31.34±0.62)%,(16.62±0.97)%演。②阴性对照未见转化生长因子β1mRNA阳性表达。甲基强的松龙组和损伤对照组均有转化生长因子β1mRNA阳性表达,甲基强的松龙组同损伤对照组相比,各时段转化生长因子β1mRNA表达均有减少,且差异明显。结论:甲基强的松龙能抑制大鼠脊髓损伤后转化生长因子β1mRNA的表达,说明甲基强的松龙对脊髓损伤的治疗作用不是通过转化生长因子β1来实现的。更多还原

【Abstract】 AIM: To investigate the effects of methylprednisolone (MP) on the expression changes of transforming growth factor-beta 1 mRNA (TGF-β1 mRNA) in rats after spinal cord injury (SCI). METHODS: The experiment was carried out in Shanghai Medical Industry Academe and Pathology Laboratory of Changhai Hospital, the Second Military Medical University of Chinese PLA from June 2003 to March 2004.Totally 45 male SD rats were divided at random into 3 groups: Blank controlled group (n=5), SCI group (6 hours, 24 hours, 3 days, 7 days) and MP group (6 hours, 24 hours, 3 days, 7 days), 5 rats in each time point. After the establishment of SCI models, the normal salines were given in the blank group and SCI group while 30 mg/kg MP in MP group. At 6 hours, 24 hours, 3 days, 7 days after the injury, the spinal cord tissues were obtained from the injured spots and approaching locus, and dealed with in situ hybridization, stained with diaminobenzidine and smeared with hematoxylin. Five high-expression sights were selected randomly and observed with light microscope, high power microscopic (200), then the percentage of positive cells (number of positive cells/total number of cells) in each high-power field was recorded. The cellular plasm showing buffy was the expression of TGF-β1 mRNA. The effects of MP on the expression of TGF-β1 mRNA in SCI rats were also observed.RESULTS: All the 45 rats were entered into the result analysis.①The number of positive cells expressed in TGF-β1 mRNA were significantly lower in the MP group than in SCI group At 6 hours, 24 hours, 3 days, 7 days after the injury, SCI group was (8.91±0.82)%, (28.27±1.10)%, (42.09±0.93)%, (45.24±0.85)% while MP group was (7.93±0.40)%, (20.36±1.10)%, (31.34±0.62)%, (16.62±0.97)% respectively. ②The positive expression of TGF-β1 mRNA were not found in the negative control. There were TGF-β1 mRNA expressions in MP group and SCI group. Compared with SCI group, the expression of TGF-β1 mRNA of MP group decreased in each time point, with the significant difference.CONCLUSION:MP can decrease the expression of TGF-β1 mRNA in rats after SCI, which indicates MP is not effective in the treatment of SCI through TGF-β1 mRNA.更多还原