摘要
目的 观察氢饱和生理盐水(HRS)对于急性坏死性胰腺炎(ANP)大鼠肝脏活性氧自由基(ROS)激活的丝裂原活化蛋白激酶(MAPKs)通路的影响,并探讨其对于ANP大鼠肝脏的保护作用.方法 90只雄性Wistar大鼠随机分为假手术(SO)组、ANP组和HRS组,每组分为3、12、24h3个亚组(n=10).逆行胆胰管注射5%牛磺胆酸钠制备ANP模型,SO组仅开腹翻动胰十二指肠后关腹.HRS组术后尾静脉补充HRS 6 ml/kg,同时皮下注射HRS 20 ml/kg,其余各组同等剂量给予生理盐水.术后分组剖杀,检测血清血淀粉酶(AMY)、脂肪酶(LIP)、谷丙转氨酶(ALT)及谷草转氨酶(AST).取胰头及肝组织固定切片,观察病理改变并评分或分级,免疫组织化学检测肝脏组织中磷酸化c-jun氨基端激酶(p-JNK)、p-p38及磷酸化细胞外信号调节激酶(p-ERK)表达.检测新鲜肝脏组织中超氧化物歧化酶(SOD)活性及丙二醛(MDA)水平.结果 各时间点ANP大鼠血清AMY(U/L)(5 385±572比1 146±121)、LIP(U/L)(1 924.9±105.4比86.5±8.9)、ALT (U/L)(156.9±15.8比56.1±5.4)、AST (U/L)(448.5±53.0比124.6±14.7)以及胰腺肝脏病理评分[(13.79±1.36)比(0.38±0.53)分]较SO组均显著升高,差异有统计学意义(P<0.05);HRS组AMY(U/L)(5 567±512比5 385 ±572)、LIP (U/L)(1 856.8±149.3比1 924.9±105.4)较ANP组下降,但差异无统计学意义(P>0.05).HRS组ALT(U/L)(134.8±11.8比156.9±15.8)、AST(U/L)(326.6±20.5比448.5±53.0)及肝脏的病理分级(17.1比23.1)较ANP组有显著降低,差异有统计学意义(P<0.05);ANP组肝脏组织SOD(U/mg)(127.2±11.6比267.4±33.2)活性水平较SO组有明显降低,MDA (nmol/mg)(12.6±3.1比3.9±1.7)水平明显升高,差异有统计学意义(P<0.05),HRS组肝脏组织中SOD (U/mg)(177.4±12.5比127.2±11.6)活性较ANP组有显著升高,MDA(nmoL/mg)(9.7±2.3比12.6±3.1)含量显著降低,差异有统计学意义(P<0.05).ANP组肝脏组织中p-JNK(121.40±10.70比2.60±0.45),p-p38(63.1±6.9比16.2±1.7),p-ERK(7.10±1.40比0.90±0.04)水平比较于SO组明显升高,差异有统计学意义(P<0.05),HRS组p-JNK(51.30±9.70比121.40±10.70)及p-p38(16.2±1.7比63.1±7.0)水平比较于ANP组有显著下降,差异有统计学意义(P<0.05),但p-ERK(6.70±1.30比7.10±1.40)表达差异无统计学意义(P>0.05).结论 HRS可有有效的降低ANP大鼠肝脏氧化应激,从而减少ROS对JNK及p38通路的激活,从而对ANP大鼠肝脏起到保护作用.
Objective To investigate the impact of hydrogen-rich saline on mitogen-activated protein kinases (MAPKs) pathway in hepatic reactive oxygen species (ROS) activation in acute necrotizing pancreatiis (ANP) and protective effect of hydrogen-rich saline on hepatic injury of ANP in rat.Methods Ninety male Wistar rats were randomly divided into three groups:sham operation group (SO group),ANP group and hydrogen-rich saline treatment group (HRS group),each group further divided into three (3,12,24 h) subgroup (n =10).ANP model was induced by retrograde infusion of 5% sodium taurocholate into the biliopancreatic duct.Hydrogen-rich saline was injected through the tail vein (6 ml/kg) and under the skin (20 ml/kg) at 5 minutes after the operation in HRS group.The rats of SO group and ANP group were administered by intravenous (6 ml/kg) and subcutaneous (20 ml/kg) injection with saline.Rats in each group were sacrificed at 3,12 and 24 h after the operation.The levels of serum amylase (AMY),lipase (LIP),alanine transaminase (ALT),aspartate aminotransferase (AST) and histopathological scores or grades of pancreatic and liver tissues were measured.Hepatic malondialdehyde content and superoxide dismutase activity were analyzed.Results The levels of serum AMY (U/L) (5 385 ± 572 vs.1 146±121),LIP (U/L) (1 924.9±105.4vs.86.5±8.9),ALT (U/L) (156.9±15.8 vs.56.1±5.4),AST (U/L) (448.5 ±53.0 vs.124.6 ± 14.7),and histopathological scores of pancreatic tissues (13.79 ± 1.36 vs.0.38 ±0.53) in ANP group increased significantly than SO group (P 〈0.05).serum AMY (U/L) (5 567 ±512 vs.5 385 ±572) and LIP (U/L) (1 856.8 ± 149.3 vs.1 924.9 ± 105.4) in HRS group decreased than ANP group,but no statistical significance (P 〉 0.05).serum ALT (U/L) (134.8±11.8 vs.156.9±15.8),AST (U/L) (326.6±20.5 vs.448.5±53.0) and hepatic histopathological grades (17.1 vs.23.1) in HRS group decreased significantly than ANP group (P 〈 0.05).Hepatic superoxide dismutase activity decreased in ANP group than SO group and hepatic malondialdehyde acyivity [(12.6 ± 3.1) vs.(3.9 ± 1.7) nmol/mg] increased in ANP group than SO group (P 〈 0.05).Hepatic superoxide dismutase activity [(177.4 ± 12.5) vs.(127.2 ± 11.6) U/mg] in HRS group were higher than ANP group and hepatic malondialdehyde acyivity [(9.7 ± 2.3) vs.(12.6 ± 3.1) nmol/mg]lower significantly than ANP group (P 〈 0.05).Hepatic phosphorylated c-Jun N-terminal kinase [p-JNK (121.40 ± 10.70 vs.2.60 ± 0.45)],p-p38 (63.1 ± 6.9 vs.16.2 ± 1.7),phosphorylated extracellular signal-regulated kinase [p-ERK (7.10 ± 1.40 vs.0.90 ± 0.04)] expression in ANP group increased significantly than SO group (P 〈 0.05).in HRS group,Hepatic p-JNK (51.30 ± 9.70 vs.121.40 ± 10.70),p-p38 (16.2 ± 1.7 vs.63.1 ± 7.0) expression decreased significantly than in ANP group (P 〈 0.05),but the p-ERK (6.70 ± 1.30 vs.7.10 ± 1.40) expression was no significance (P 〉 0.05).Conclusion Hydrogen-rich saline may effectively decrease hepatic oxidative stress in ANP rat and decrease ROS activation of JNK and p38 pathway.By this way,has a protective effect for hepatic injury of ANP rat.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2015年第1期33-36,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(81370562)
湖北省自然科学基金资助项目(2013CFB459)
湖北省卫生厅科研基金重点资助项目(JX6A07)
关键词
胰腺炎
氢饱和生理盐水
肝损伤
氧化应激
丝裂原活化蛋白激酶
Pancreatitis
Hydrogen-rich saline
Hepatic injury
Oxidative stress
Mitogen-activated protein kinases