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目的探讨髓样细胞分化蛋白88(MyD88)在重症急性胰腺炎(SAP)发病机理的作用。方法将48只小鼠按随机数字表法随机分为SAP组(32只)与正常对照组(16只);再将2组小鼠随机(随机数字表法)分为6、12、24及48 h组,SAP组各亚组每组8只,正常对照组每亚组4只。SAP组小鼠腹腔注射20%L-精氨酸以诱导SAP模型,正常对照组小鼠仅腹腔注射生理盐水。分别于建模术后6、12、24及48 h处死小鼠,取其动脉血,采用ELISA方法检测血清中白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)及肿瘤坏死因子-α(TNF-α)浓度;同时取其胰腺组织(正常对照组仅术后6 h取材),用逆转录-聚合酶链反应(RT-PCR)方法检测胰腺组织中MyD88 mRNA和核因子-κB(NF-κB)mRNA的表达水平,并进行HE染色。结果镜下见SAP组小鼠的胰腺组织随时间进展其炎症逐渐加重。各时点SAP组小鼠的IL-1β、IL-10及TNF-α浓度均高于正常对照组(P<0.05);各时点SAP组与正常对照组(6 h组)相比较,其胰腺组织中MyD88 mRNA及NF-κB mRNA的表达水平均较高(P<0.05)。各时点SAP组小鼠MyD88 mRNA的表达水平与血清IL-1β、IL-10及TNF-α的浓度和NF-κB mRNA的表达水平均呈正相关(P<0.01)。结论 MyD88的表达对SAP的发生和发展可能均具有重要的作用。
Objective To investigate the role of MyD88 in the pathogenesis of severe acute pancreatitis (SAP). Methods Forty-eight mice were randomly divided into SAP group (n = 32) and normal control group (n = 16) according to random number table method. Randomly divided two groups of mice (random number table) into 6,12,24 and 48 h group, SAP group each subgroup 8 each, normal control group 4 subgroups. The SAP group mice were intraperitoneally injected with 20% L-arginine to induce SAP model, while the normal control group mice were injected intraperitoneally with normal saline. The mice were sacrificed at 6, 12, 24 and 48 h after modeling respectively. The arterial blood samples were collected for determination of interleukin-1β (IL-1β), interleukin-10 (IL- And tumor necrosis factor-α (TNF-α) were also detected. At the same time, their pancreatic tissues were taken from the normal control group (only 6 h after operation). The expression of MyD88 mRNA in pancreatic tissue was detected by reverse transcription polymerase chain reaction And nuclear factor-κB (NF-κB) mRNA expression levels, and HE staining. Results Microscopically, the pancreas tissue of SAP mice gradually increased with time. The concentrations of IL-1β, IL-10 and TNF-α in SAP group were higher than those in normal control group at each time point (P <0.05). Compared with normal control group (6 h group) The expression of MyD88 mRNA and NF-κB mRNA in pancreatic tissue were higher (P <0.05). The expression level of MyD88 mRNA in SAP group was positively correlated with the concentration of IL-1β, IL-10 and TNF-α and the expression of NF-κB mRNA at each time point (P <0.01). Conclusion The expression of MyD88 may play an important role in the occurrence and development of SAP.