己酮可可碱具有抗大鼠非酒精性脂肪性肝炎的作用

Pentoxifylline have anti-inflammatory effect on nonalcoholic steatohepatitis in rats

目的探讨己酮可可碱(PTX)对非酒精性脂肪性肝炎(NASH)大鼠肝脏的抗炎作用。方法SD大鼠40只,标准饲料喂养1周后,随机分为4组:对照组、12周模型组、16周模型组和PTX治疗组。用高脂饮食法建立非酒精性脂肪性肝炎模型,16周末处死全部大鼠,收集血浆和肝组织,采用酶联免疫吸附法(ELISA)检测血浆肿瘤坏死因子α(TNF-α)的浓度,放射免疫法(RI)检测血浆白细胞介素1β(IL-1β)、白细胞介素6(IL-6)的浓度,采用免疫组织化学观察肝脏白细胞介素1β(IL-1β)、白细胞介素6(IL-6)的表达,采用电泳迁移率实验(EMSA)检测肝组织核转录因子-κB(NF-κB)的活性变化,RT-PCR方法检测肝脏TNF-αmRNA的转录。结果模型组血浆TNF-α,IL-1β,IL-6均较正常组有所升高,而治疗组较模型组有所降低。免疫组织化学实验结果表明:与对照组相比,12周,16周模型组及PTX治疗组肝脏IL-1β,IL-6的表达显著增加(P<0.05),与12周模型组相比,PTX治疗组IL-1β,IL-6的表达有所降低(P<0.05)。与16周模型组相比,PTX治疗组IL-6的表达有所降低(P<0.05)。EMSA结果显示:与对照组相比,模型组NF-κB的活性逐渐增高,经治疗后,PTX治疗组NF-κB的活性有所减低。PT-PCR检测结果显示,与对照组相比,16周模型组TNF-αmRNA的表达有显著的升高(P<0.05)。与16周模型组相比,PTX治疗组TNF-αmRNA的表达有显著的降低(P<0.05)。结论PTX对NASH具有一定的抗炎作用。

Objective To study the effects of pentoxifylline （PTX） on anti-inflammatory factor of nonalcoholic steatohepatitis （NASH） in rats. Methods Rats fed a standard diet for one week were randomly divided into four groups, control group（ n = 10）, 12 weeks model group（ n = 10）, 16 weeks model group（ n = 10） and the PTX treatment group（ n = 10）. Two model groups and the PTX treatment group were then fed a high-fat diet. Meanwhile the rats fed a standard diet served as control group. After 12 weeks, the 12 weeks model group were sacrificed and the treatment group were given PTX [16mg/（kg·d）] by intragastric administration. Then all the rats of other three groups were sacrificed at the end of the 16th week. Blood samples were collected aseptically from the abdominalis aorta and serum was separated with regular methods. Serum level of tumour necrosis factor a（TNF-α）was determined by ELISA. Serum levels of IL-1β and IL-6 were tested by radio immunoassay（RI）. The immunohistochemical staining was perfored on the liver tissue slice, the expressions of IL-1β, IL-6 were evaluated. TNF-α mRNA was detected by RT-PCR. NF-κB binding activity was detected by electrophoretic mobility shift assay（EMSA）. Results Serum level of TNF-α, IL-1β and IL-6 of model groups increased significantly than that of the control group. No statistical difference was found among the groups. The expressions of IL-1β,IL-6 were increased significantly in the model groups and the PTX treatment group （vs. the control group, P 〈 0.05）, while the expression of IL-6 was decreased in the PTX treatment group （vs. the model groups）; The level of TNF-α mRNA was increased significantly in the 16 weeks model groups （vs. the control group, P 〈 0.05）, while decreased in the PTX treatment group （vs. the 16 weeks model groups, P 〈 0.05）. Conclusion PTX can relieve the levels of inflammatory factors, and exert anti-inflammatory effect.