中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2651-2654
,共4页
赵亮%石乔%左腾%邓文宏%王卫星
趙亮%石喬%左騰%鄧文宏%王衛星
조량%석교%좌등%산문굉%왕위성
氢饱和生理盐水%急性坏死性胰腺炎%肝损伤%c-jun氨基端激酶%凋亡
氫飽和生理鹽水%急性壞死性胰腺炎%肝損傷%c-jun氨基耑激酶%凋亡
경포화생리염수%급성배사성이선염%간손상%c-jun안기단격매%조망
Hydrogen-rich saline%Acute necrotizing pancreatitis%Liver injury%c-Jun N-terminal kinase%Apoptosis
目的 探讨氢饱和生理盐水(HRS)对急性坏死性胰腺炎(ANP)肝损伤的治疗作用及其可能机制.方法 将72只雄性Wistar大鼠随机(随机数字法)分为假手术(SO)组、ANP组和HRS组,每组24只.采用胆胰管逆行注射5％牛磺胆酸钠方法制备ANP模型.HRS组在造模成功后5 min尾静脉注射HRS 6 ml/kg体质量,并皮下注射HRS 20 ml/kg体质量补液.SO组大鼠开腹后仅翻动十二指肠和胰腺后关腹.SO组和ANP组大鼠在术后5 min经尾静脉注射生理盐水(6ml/kg体质量),并皮下注射生理盐水(20 ml/kg体质量)补液.术后3、12、24 h分批处死大鼠,心脏取血检测血清淀粉酶(AMY)、谷丙转氨酶(ALT)及谷草转氨酶(AST)水平.取胰头及肝左叶组织行病理检查并评分/分级.免疫组织化学法检测各组大鼠肝脏组织中c-jun氨基端激酶(JNK)、磷酸化c-jun氨基端激酶(p-JNK)蛋白的表达.脱氧核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测肝脏细胞凋亡.结果 ANP组各时间点的AMY、ALT、AST、胰腺及肝脏病理评分/分级均较SO组大鼠显著升高(P ＜0.05);HRS组相应指标数值较ANP组均显著下降,但仍显著高于SO组(P＜0.05).ANP组12h大鼠肝脏细胞胞质中JNK大量表达,p-JNK广泛表达于肝细胞的胞质及细胞核内;HRS组对应时间点大鼠肝细胞内JNK、p-JNK表达显著低于ANP组(P＜0.05).ANP组12h点大鼠肝脏组织凋亡细胞在总细胞的所占比例[(68.78±5.63)％]与SO组[(15.46±5.32)％]比较显著升高(t=19.472,P＜0.05),HRS组大鼠肝脏凋亡细胞比例[(36.99±7.42)％]与ANP组比较显著降低(t=-9.65,P＜0.05).结论 HRS对ANP大鼠肝损伤具有显著治疗作用,这种作用可能是通过降低JNK表达及磷酸化水平,进而减少肝细胞凋亡来实现的.
目的 探討氫飽和生理鹽水(HRS)對急性壞死性胰腺炎(ANP)肝損傷的治療作用及其可能機製.方法 將72隻雄性Wistar大鼠隨機(隨機數字法)分為假手術(SO)組、ANP組和HRS組,每組24隻.採用膽胰管逆行註射5％牛磺膽痠鈉方法製備ANP模型.HRS組在造模成功後5 min尾靜脈註射HRS 6 ml/kg體質量,併皮下註射HRS 20 ml/kg體質量補液.SO組大鼠開腹後僅翻動十二指腸和胰腺後關腹.SO組和ANP組大鼠在術後5 min經尾靜脈註射生理鹽水(6ml/kg體質量),併皮下註射生理鹽水(20 ml/kg體質量)補液.術後3、12、24 h分批處死大鼠,心髒取血檢測血清澱粉酶(AMY)、穀丙轉氨酶(ALT)及穀草轉氨酶(AST)水平.取胰頭及肝左葉組織行病理檢查併評分/分級.免疫組織化學法檢測各組大鼠肝髒組織中c-jun氨基耑激酶(JNK)、燐痠化c-jun氨基耑激酶(p-JNK)蛋白的錶達.脫氧覈苷痠末耑轉移酶介導的缺口末耑標記(TUNEL)法檢測肝髒細胞凋亡.結果 ANP組各時間點的AMY、ALT、AST、胰腺及肝髒病理評分/分級均較SO組大鼠顯著升高(P ＜0.05);HRS組相應指標數值較ANP組均顯著下降,但仍顯著高于SO組(P＜0.05).ANP組12h大鼠肝髒細胞胞質中JNK大量錶達,p-JNK廣汎錶達于肝細胞的胞質及細胞覈內;HRS組對應時間點大鼠肝細胞內JNK、p-JNK錶達顯著低于ANP組(P＜0.05).ANP組12h點大鼠肝髒組織凋亡細胞在總細胞的所佔比例[(68.78±5.63)％]與SO組[(15.46±5.32)％]比較顯著升高(t=19.472,P＜0.05),HRS組大鼠肝髒凋亡細胞比例[(36.99±7.42)％]與ANP組比較顯著降低(t=-9.65,P＜0.05).結論 HRS對ANP大鼠肝損傷具有顯著治療作用,這種作用可能是通過降低JNK錶達及燐痠化水平,進而減少肝細胞凋亡來實現的.
목적 탐토경포화생리염수(HRS)대급성배사성이선염(ANP)간손상적치료작용급기가능궤제.방법 장72지웅성Wistar대서수궤(수궤수자법)분위가수술(SO)조、ANP조화HRS조,매조24지.채용담이관역행주사5％우광담산납방법제비ANP모형.HRS조재조모성공후5 min미정맥주사HRS 6 ml/kg체질량,병피하주사HRS 20 ml/kg체질량보액.SO조대서개복후부번동십이지장화이선후관복.SO조화ANP조대서재술후5 min경미정맥주사생리염수(6ml/kg체질량),병피하주사생리염수(20 ml/kg체질량)보액.술후3、12、24 h분비처사대서,심장취혈검측혈청정분매(AMY)、곡병전안매(ALT)급곡초전안매(AST)수평.취이두급간좌협조직행병리검사병평분/분급.면역조직화학법검측각조대서간장조직중c-jun안기단격매(JNK)、린산화c-jun안기단격매(p-JNK)단백적표체.탈양핵감산말단전이매개도적결구말단표기(TUNEL)법검측간장세포조망.결과 ANP조각시간점적AMY、ALT、AST、이선급간장병리평분/분급균교SO조대서현저승고(P ＜0.05);HRS조상응지표수치교ANP조균현저하강,단잉현저고우SO조(P＜0.05).ANP조12h대서간장세포포질중JNK대량표체,p-JNK엄범표체우간세포적포질급세포핵내;HRS조대응시간점대서간세포내JNK、p-JNK표체현저저우ANP조(P＜0.05).ANP조12h점대서간장조직조망세포재총세포적소점비례[(68.78±5.63)％]여SO조[(15.46±5.32)％]비교현저승고(t=19.472,P＜0.05),HRS조대서간장조망세포비례[(36.99±7.42)％]여ANP조비교현저강저(t=-9.65,P＜0.05).결론 HRS대ANP대서간손상구유현저치료작용,저충작용가능시통과강저JNK표체급린산화수평,진이감소간세포조망래실현적.
Objective To investigate the therapeutic effect and it's mechanism of hydrogen rich saline on liver injury in rats with acute necrotizing pancreatitis.Methods Seventy-two male Wistar rats were randomly divided into three groups: sham operation roup (SO group), acute nccrotizing pancreatitis group (ANP group) and hydrogen-rich saline treatment group (HRS group), with 24 rats in each group.Acute necrotizing pancreatitis model was induced by retrograde injection of 5％ sodium taurocholate into the cholangiopancreatic duct.Hydrogen-rich saline was injected through the tail vein (6 ml/kg) and subcutaneously (20 ml/kg) at 5 minutes after the operation in HRS group.The rats of SO group underwent laparotomy with gentle pancreas, duodenum manipulation only.The rats of SO group and ANP group were administered with 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, alanine transaminase (ALT) and aspartate aminotransferase (AST) were determined.Pathological changes of liver and pancreas tissues were examined and pathological scores/grades were recorded.The expression of c-Jun N-terminal kinase (JNK), phosphorylated c-Jun N-terminal kinase (p-JNK) were detected by immunohistochemical technology.The apoptosis of liver were examined by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay.Results The levels of serum amylase (AMY), ALT, AST, liver and pancreas immunohistochemical scores in ANP group were significantly higher than those in SO group (P ＜ 0.05).The corresponding values in HRS group were significantly lower than those in ANP group (P ＜ 0.05).JNK was determined in the cytoplasm and p-JNK was detected in nucleus ANP group,and their expression intensities were dramatically higher than that of SO group and HRS group (P ＜ 0.05).The proportion of TUNEL-positive cells in ANP group [(68.78 ± 5.63) ％] at 12 h were notably higher that of SO group [(15.46 ±5.32)％, t=19.472, P＜0.05];but the rate in HRS group [(36.99 ± 7.42) ％] at corresponding time were markedly lower than that of ANP group (t =-9.65, P ＜ 0.05).Conclusion Hydrogen-rich saline has remarkable therapeutic effect on liver injury of ANP, and its mechanism may be related to reduction of apoptosis of liver cells, which caused by the overexpression and phosphorylation of JNK.
