【作者】 方勇木；

【导师】 黄鹤光；

【作者基本信息】 福建医科大学 ， 外科学， 2009， 硕士

【摘要】 目的:探讨p38丝裂原活化蛋白激酶(p38MAPK)信号转导通路对重症急性胰腺炎(SAP)大鼠低钙血症和甲状旁腺激素受体1(PTHR1)表达的影响。方法:将雄性SD大鼠72只随机分为:SAP组;治疗组(SB组);假手术组(SO组),每组分3h,6h,12h三个时间点,每个时间点8只。以5%牛磺脱氧胆酸钠逆行胰胆管注射建立SAP模型,SB组在造模前30分钟腹腔注射p38MAPK特异抑制剂SB203580。观察各组血清钙浓度,蛋白免疫印迹技术(Western blot)分析骨组织磷酸化p38MAPK(P-p38MAPK)和肿瘤坏死因子-α(TNF-α)变化,实时逆转录聚合酶链反应(Real-time RT-PCR)检测骨组织PTHR1mRNA表达。结果:SAP组与SO组比较骨组织P-p38MAPK和TNF-α的表达量明显增高,骨组织PTHR1mRNA表达量和血清钙浓度显著下降。SB组与SAP组比较骨组织P-p38MAPK [3h:(0.20±0.05)vs.( 0.40±0.06),t=-7.044,P=0.000;6h:(0.33±0.05)vs.(0.80±0.06),t=-18.099,P=0.000;12h:(0.27±0.05)vs.(0.51±0.03),t=-12.385,P=0.000]和TNF-α[3h:(0.31±0.03)vs.(0.70±0.03),t=-23.870,P=0.000;6h:(0.44±0.03)vs.(0.91±0.04),t=-27.755,P=0.000;12h:(0.37±0.04)vs.(0.80±0.03),t=-23.652,P=0.000]表达显著下调,骨组织PTHR1mRNA表达量[3h:(0.53±0.03)vs.( 0.28±0.03),t=17.858,P=0.000;6h:(0.44±0.06)vs.( 0.23±0.04),t=8.804,P=0.000;12h:(0.40±0.11)vs.( 0.20±0.03),t=5.060,P=0.001]和血清钙浓度[3h:(2.39±0.06)mmol/L vs.( 2.31±0.06) mmol/L,t=2.900,P=0.012;6h:(2.35±0.10) mmol/L vs.( 2.11±0.06) mmol/L,t=5.507,P=0.000;12h:(2.26±0.12) mmol/L vs.( 2.01±0.04) mmol/L,t=5.881,P=0.000]显著升高。结论:p38MAPK信号转导通路可介导SAP低钙血症的发生,抑制该通路可改善SAP低钙血症。更多还原

【Abstract】 Objective To investigate the role of p38 mitogen-activated protein kinase(MAPK) signal transduction pathway in hypocalcaemia and parathyroid hormone receptor 1 (PTHR1) of rats with severe acute pancreatitis (SAP). Methods Seventy-two male health adult Sprague-dawley rats were completely randomized into three groups:SAP group,SAP treated with SB203580 group (SB group),sham operation group(SO group).SAP model was induced by bili-pancreatic duct retrograde infusion with 5% sodium taurocholate solution.In the SB group,rats were treated with the specific inhibior (SB203580) of p38MAPK 30 minutes before the induction of SAP model. Rats in each group were killed at 3h,6h,12h after operation to determine the Serum calcium, the change of phosphorylated p38MAPK and tumor necrosis factor (TNF)-alpha (by western blot) and the expression of PTHR1mRNA(by quantitative real-time RT-PCR),8 rats in each time point.Results Compared with sham group,the expression of bone phosphorylated p38MAPK and TNF-alpha were increased significantly,the mRNA level of bone PTHR1 down-regulated significantly and the concentration of serum calcium decreased significantly in SAP group.Compared with SAP group,the expression of bone phosphorylated p38MAPK [3h:(0.20±0.05)vs.( 0.40±0.06),t=-7.044,P=0.000;6h:(0.33±0.05)vs.(0.80±0.06),t=-18.099,P=0.000;12h:(0.27±0.05)vs.(0.51±0.03),t=-12.385,P=0.000] and TNF-alpha [3h:(0.31±0.03)vs.(0.70±0.03),t=-23.870,P=0.000;6h:(0.44±0.03)vs.(0.91±0.04),t=-27.755,P=0.000;12h:(0.37±0.04)vs.(0.80±0.03),t=-23.652,P=0.000] were decreased significantly, the mRNA level of bone PTHR1 [3h:(0.53±0.03)vs.( 0.28±0.03),t=17.858,P=0.000;6h:(0.44±0.06)vs.( 0.23±0.04),t=8.804,P=0.000;12h:(0.40±0.11)vs.( 0.20±0.03),t=5.060,P=0.001] up-regulated significantly and the concentration of serum calcium [3h:(2.39±0.06) mmol/L vs.( 2.31 ±0.06) mmol/L,t=2.900,P=0.012;6h:(2.35±0.10) mmol/L vs.( 2.11±0.06) mmol/L,t=5.507,P=0.000;12h:(2.26±0.12) mmol/L vs.( 2.01±0.04) mmol/L,t = 5.881,P = 0.000] increased significantly in SB group.Conclusion P38MAPK signal transduction pathway may mediates the development of hypocalcaemia,the level of serum calcium could be increased by blocking this pathway in rats with SAP.更多还原