CAJ | 학술논문

目的探讨急性胰腺炎大鼠脑组织中TNF-α mRNA和蛋白的表达及其对胰性脑病发病机制的意义.方法应用5%、1%的牛磺胆酸钠分别诱导急性坏死性胰腺炎(ANP)和急性水肿性胰腺炎(AEP)大鼠模型,同时设立假手术组,每组16只.分别于模型诱导后12 h,以酶联免疫吸附(ELISA)法测定大鼠血液和脑脊液标本中TNF-α的含量,RT-PCR方法测定脑组织TNF-α mRNA的表达,Western-Blot测定TNF-α蛋白的表达水平,免疫组化观察TNF-α蛋白的定位表达.结果 ANP组、AEP组和假手术组的血清TNF-α水平分别为1065.6pg/ml、577.5pg/ml和543.8pg/ml,脑脊液TNF-α水平分别为820.9pg/ml、307.4pg/ml和481.6pg/ml.ANP组血清和脑脊液TNF-α水平同AEP组和假手术组相比,差异均有显著性(P＜0.05).ANP组脑组织TNF-α mRNA和蛋白的表达明显增强.ANP组TNF-α蛋白的表达定位于大脑小胶质细胞和神经元细胞.结论在ANP病程中,大鼠脑组织可能是细胞因子过度表达的来源之一;脑组织中,TNF-α主要由小胶质细胞和神经元细胞表达;TNF-α的过度表达可能是ANP早期脑髓鞘损伤的原因之一.
목적 탐토급성이선염대서뇌조직중TNF-α mRNA화단백적표체급기대이성뇌병발병궤제적의의.방법 응용5%、1%적우광담산납분별유도급성배사성이선염(ANP)화급성수종성이선염(AEP)대서모형,동시설립가수술조,매조16지.분별우모형유도후12 h,이매련면역흡부(ELISA)법측정대서혈액화뇌척액표본중TNF-α적함량,RT-PCR방법측정뇌조직TNF-α mRNA적표체,Western-Blot측정TNF-α단백적표체수평,면역조화관찰TNF-α단백적정위표체.결과 ANP조、AEP조화가수술조적혈청TNF-α수평분별위1065.6pg/ml、577.5pg/ml화543.8pg/ml,뇌척액TNF-α수평분별위820.9pg/ml、307.4pg/ml화481.6pg/ml.ANP조혈청화뇌척액TNF-α수평동AEP조화가수술조상비,차이균유현저성(P＜0.05).ANP조뇌조직TNF-α mRNA화단백적표체명현증강.ANP조TNF-α단백적표체정위우대뇌소효질세포화신경원세포.결론 재ANP병정중,대서뇌조직가능시세포인자과도표체적래원지일;뇌조직중,TNF-α주요유소효질세포화신경원세포표체;TNF-α적과도표체가능시ANP조기뇌수초손상적원인지일.
Objective To investigate the expression of TNF-α messenger RNA(mRNA)and peptide in the brain of rats with acute pancreatitis(AP)and explore its significance in the pathogenesis of pancreatic encephalopathy(PE).Methods Modds of acute necrotizing pancreatitis(ANP,n=16)and acute edematous pancreatitis(AEP,n=16)were induced by injecting sodium taurocholate into the pancreatic duct of male Sprague Dawley rats.Another 16 normal rats were employed to serve as controls.Twelve hours after the induction of AP,the rats in each group were sacrificed and the blood and cerebral spinal fluid(CSF)samples were taken for determining the level of TNF-α by ELISA.TNF-α mRNA and peptide expression in the cerebral tissues was assessed by semiquantitative RT-PCR,Western blotting and immunohistochemistry,respectively.Results TNFF-α concentration in blood and CSF of rats with ANP showed significant elevation as compared with AEP and control grouP(P＜0.05).TNF-α concentration in blood of ANP,AEP and control group were 1065.6pg/ml,577.5pg/ml and 543.8pg/ml,respectively.The concentration in CSF was 820.9pg/ml,307.4pg/ml and 481.6Pg/ml,respectively.TNF-α mRNA expression and its protein level in the cerebra of rats with ANP were significantly greater than that in the other two groups respectively(P＜0.05).Double-labeling and immunohistochemical studies revealed that the protein was located in the microglial cells and neurons in the cerebrum.Conclusion The cerebrum might be one of the organs releasing cytokines during AP.TNF-α is overproduced by both microglial cells and neurons in the cerebrunl and might play an important role in the demyelinization of brain during the early stage of ANP.