CAJ | 학술논문

目的：探讨褪黑素（MT）对脊髓损伤（SCI）大鼠体内肿瘤坏死因子-α（TNF-α）表达的影响及可能机制。方法：108只SD大鼠随机分为3组。假手术（Sham）组仅行椎板切除不损伤脊髓，SCI组与SCI＋MT组采用改良的Allen’s法建立SCI模型；SCI＋MT组术后10 min腹腔注射MT（100 mg/kg），SCI组注射等量体积分数5％乙醇。术后12 h各组选6只取脊髓，采用HE染色法和免疫组化法分别观察脊髓组织的病理变化和TNF-α蛋白的表达情况。术后1、12、24、48、72 h，各组分别取6只大鼠，用ELISA法检测血清TNF-α水平， RT-PCR法测定脊髓组织TNF-αmRNA的表达。结果：SCI＋MT组病理改变较SCI组明显减轻。脊髓组织中TNF-α蛋白主要分布在神经元和胶质细胞的胞浆和胞核；3组TNF-α阳性细胞数差异有统计学意义（F＝504．239，P＜0．001），其中SCI组最高， SCI＋MT组次之，Sham组最低。各时间点3组血清TNF-α水平和脊髓组织中TNF-αmRNA的表达水平差异有统计学意义（P均＜0．05），其中SCI＋MT组上述2指标低于SCI组，但仍高于Sham组（P＜0．05）。结论：MT可能通过抑制TNF-α的表达水平减轻损伤脊髓的继发性炎症损伤。
목적：탐토퇴흑소（MT）대척수손상（SCI）대서체내종류배사인자-α（TNF-α）표체적영향급가능궤제。방법：108지SD대서수궤분위3조。가수술（Sham）조부행추판절제불손상척수，SCI조여SCI＋MT조채용개량적Allen’s법건립SCI모형；SCI＋MT조술후10 min복강주사MT（100 mg/kg），SCI조주사등량체적분수5％을순。술후12 h각조선6지취척수，채용HE염색법화면역조화법분별관찰척수조직적병리변화화TNF-α단백적표체정황。술후1、12、24、48、72 h，각조분별취6지대서，용ELISA법검측혈청TNF-α수평， RT-PCR법측정척수조직TNF-αmRNA적표체。결과：SCI＋MT조병리개변교SCI조명현감경。척수조직중TNF-α단백주요분포재신경원화효질세포적포장화포핵；3조TNF-α양성세포수차이유통계학의의（F＝504．239，P＜0．001），기중SCI조최고， SCI＋MT조차지，Sham조최저。각시간점3조혈청TNF-α수평화척수조직중TNF-αmRNA적표체수평차이유통계학의의（P균＜0．05），기중SCI＋MT조상술2지표저우SCI조，단잉고우Sham조（P＜0．05）。결론：MT가능통과억제TNF-α적표체수평감경손상척수적계발성염증손상。
Aim:To investigate the effects of melatonin ( MT) on the expression of tumor necrosis factor-α( TNF-α) in acute spinal cord injury ( SCI) rats.Methods:A total of 108 SD rats were randomly allocated into three groups:Sham group only accepted laminectomy without SCI , and SCI group and SCI +MT group used the modified Allen's method to establish SCI model.100 mg/kg MT was given to the rats in SCI +MT group, and 5% ethanol, to SCI group at 10 min after the model establishment .At 12 h, spinal cord samples of 6 rats from each group were prepared for histological examination u-sing HE and immunohistochemical SP staining to detect the expression of TNF-α.At 1,12,24,48 and 72 h, 6 rats in each group were executed ,the serum level of TNF-αwas determined by ELISA , and the expression of TNF-αmRNA was detec-ted by RT-PCR.Results:Compared with SCI group ,the pathological changes in SCI +MT group were significantly allevia-ted.Immunohistochemical staining showed that TNF-αprotein in spinal cord tissue mainly located in the cytoplasm and nu-cleus of neurons and glial cells;SCI group was the highest ,SCI+MT group followed , and Sham group was the lowest ( F=504.239,P<0.001).At different time points, the serum level of TNF-αand the expression of TNF-αin spinal cord tissue in SCI+MT group were significantly lower than those in SCI group , but still higher than those in Sham group (P<0.05). Conclusion:MT may reduce secondary injury of SCI by inhibiting the expression of TNF-α.