CAJ | 학술논문

目的观察急性坏死性胰腺炎( ANP)大鼠胰腺组织水通道蛋白1(AQP1)的表达以及大承气汤对其的影响.方法 160只雄性SD大鼠按随机数字法分为对照组、ANP组、地塞米松组、乙酰唑胺组及大承气汤组,每组32只.采用胆胰管逆行注射5％牛磺胆酸钠方法制备ANP模型.地塞米松组于造模后即刻静脉给予地塞米松4 mg/kg体重；乙酰唑胺组于造模前2h用含乙酰唑胺的生理盐水1ml灌胃；大承气汤组于造模前48、24、2h分别用大承气汤2ml/次灌胃；对照组仅开腹触摸胰腺数次后关腹.制模后3、6、12、18 h分批处死8只大鼠.记录腹水量；检测血清淀粉酶；胰腺组织病理检查及电镜观察；伊文思兰(EB)血管外渗法检测毛细血管通透性；实时PCR和蛋白质印迹法检测AQP1 mRNA和蛋白表达.结果 ANP组血清淀粉酶水平显著升高,胰腺损伤明显；地塞米松组和大承气汤组淀粉酶水平较ANP组降低,胰腺损伤减轻；乙酰唑胺组淀粉酶水平高于ANP组,胰腺病理损伤较ANP组加重.造模后6h,对照组、ANP组、地塞米松组、乙酰唑胺组、大承气汤组胰腺组织EB含量分别为(13.44±2.56)、( 126.35±14.80)、(86.31±14.46)、(108.99±15.07)、(78.29±16.85) mg/L;AQP1 mRNA表达量为(170.07±22.48)％、(83.93±8.98)％、(117.09±10.70)％、(69.00±8.98)％、(112.82±11.79)％；AQP1蛋白表达量为0.23±0.06、0.10±0.02、0.32±0.03、0.13±0.02、0.45±0.04.ANP组的EB量显著高于对照组,而AQP1mRNA及蛋白的表达显著低于对照组(P值均＜0.05)；地塞米松组及大承气汤组EB含量显著低于ANP组,而AQP1 mRNA及蛋白的表达显著高于ANP组(P值均＜0.05).结论 AQP1在ANP大鼠胰腺组织毛细血管渗漏的发生中起重要作用.大承气汤通过调控AQP1的表达可减轻ANP大鼠的胰腺损伤.
목적 관찰급성배사성이선염( ANP)대서이선조직수통도단백1(AQP1)적표체이급대승기탕대기적영향.방법 160지웅성SD대서안수궤수자법분위대조조、ANP조、지새미송조、을선서알조급대승기탕조,매조32지.채용담이관역행주사5％우광담산납방법제비ANP모형.지새미송조우조모후즉각정맥급여지새미송4 mg/kg체중；을선서알조우조모전2h용함을선서알적생리염수1ml관위；대승기탕조우조모전48、24、2h분별용대승기탕2ml/차관위；대조조부개복촉모이선수차후관복.제모후3、6、12、18 h분비처사8지대서.기록복수량；검측혈청정분매；이선조직병리검사급전경관찰；이문사란(EB)혈관외삼법검측모세혈관통투성；실시PCR화단백질인적법검측AQP1 mRNA화단백표체.결과 ANP조혈청정분매수평현저승고,이선손상명현；지새미송조화대승기탕조정분매수평교ANP조강저,이선손상감경；을선서알조정분매수평고우ANP조,이선병리손상교ANP조가중.조모후6h,대조조、ANP조、지새미송조、을선서알조、대승기탕조이선조직EB함량분별위(13.44±2.56)、( 126.35±14.80)、(86.31±14.46)、(108.99±15.07)、(78.29±16.85) mg/L;AQP1 mRNA표체량위(170.07±22.48)％、(83.93±8.98)％、(117.09±10.70)％、(69.00±8.98)％、(112.82±11.79)％；AQP1단백표체량위0.23±0.06、0.10±0.02、0.32±0.03、0.13±0.02、0.45±0.04.ANP조적EB량현저고우대조조,이AQP1mRNA급단백적표체현저저우대조조(P치균＜0.05)；지새미송조급대승기탕조EB함량현저저우ANP조,이AQP1 mRNA급단백적표체현저고우ANP조(P치균＜0.05).결론 AQP1재ANP대서이선조직모세혈관삼루적발생중기중요작용.대승기탕통과조공AQP1적표체가감경ANP대서적이선손상.
Objective To detect the expression of aquaporin 1 in pancreas of rats with acute necrotizing pancreatitis (ANP) and to study the effect of Dachengqi decoction on it.Methods One hundred and sixty male SD rats were randomly divided into control group ( C group,n =32 ),ANP group ( n =32),Dexamethasone group (De group,n =32),Acetazolamide group (A group,n =32) and Dachengqi decoction group (DD group,n =32).ANP model was induced by retrograde injection of 5％ sodium taurocholate into the biliary and pancreatic duct.Rats in De group received dexamethasone (4 mg/kg) intravenously after ANP induction; while rats in A group received 1 ml acetazolamide via gastric lavage 2 h before ANP induction; rats in DD group received 2 ml Dachengqi decoction via gastric lavage 48,24,2h before ANP induction; rats in C group received laparotomy.Eight rats in each group were sacrificed at 3 h,6 h,12 h and 18h after induction of ANP models.Quantity of ascites and levels of serum amylases were measured.Pathological changes in pancreas tissue were detected by HE and electron microscope.Capillary permeability in pancreas tissue was detected by Evans Blue (EB) extravasations method.AQP1 expression in pancreas tissue was detected by real-time PCR and Western blotting.Results Levels of serum amylase in ANP group was significantly higher,and the pancreatic injuries were obvious ; the levels of serum amylase in De group and DD group was lower than that in ANP group,and the pancreatic injuries were attenuated.The levels of serum amylase in A group were higher than that in ANP group,and the pancreatic.injuries were more severe than that in ANP group.Six hours after ANP induction,the levels of EB in pancreas were (13.44 ±2.56),(126.35 ± 14.80),(86.31 ± 14.46),( 108.99 ± 15.07 ),(78.29 ± 16.85 ) mg/L In C group,ANP group,De group,A group and DD group,and the expression of AQP1 mRNA in pancreatic tissue was ( 170.07 ± 22.48 ) ％,( 83.93 ± 8.98 ) ％,( 117.09 ±10.70 ) ％,( 69.00 ± 8.98 ) ％,( 112.82 ± 11.79 ) ％ ; and the expression of AQP1 protein was 0.23 ± 0.06,0.10 ±0.02,0.32 ±0.03,0.13 ±0.02,0.45 ±0.04.The content of EB in ANP group was higher than that in C group,while the expression of AQP1 mRNA and protein in ANP group was significantly lower than that in C group (P ＜ 0.05 ).The content of EB in De group and DD group was significantly lower than that in ANP group,while the expression of AQP1 mRNA and protein was significantly higher than that in ANP group (P ＜ 0.05).Conclusions AQP1 plays an important role in the pathogenesis of capillary endothelial barrier dysfunction in rats with ANP.Dachengqi Decoction can attenuate pancreatic injuries of rats by regulating the expression of AQP1.