国际外科学杂志
國際外科學雜誌
국제외과학잡지
INTERNATIONAL JOURNAL OF SURGERY
2013年
1期
15-19,封3
,共6页
方海宗%陈志耀%黄鹤光%林葆
方海宗%陳誌耀%黃鶴光%林葆
방해종%진지요%황학광%림보
胰腺炎%环AMP依赖性蛋白激酶类%肺损伤%血管扩张刺激磷蛋白
胰腺炎%環AMP依賴性蛋白激酶類%肺損傷%血管擴張刺激燐蛋白
이선염%배AMP의뢰성단백격매류%폐손상%혈관확장자격린단백
Pancreatitis%Cyclic AMP-dependent protein kinases%Lung injury%Vasodilator-stimulated phosphoprotein
目的 探讨环磷酸腺苷/蛋白激酶A(cAMP/PKA)信号转导通路在重症急性胰腺炎肺损伤的作用机制.方法 健康雄性SD大鼠72只,按完全随机法分为假手术(SO)组、重症急性胰腺炎组(SAP组)、SAP+ H89(cAMP抑制剂)组,后两组按取材时间不同又分为3、6、12及24 h四个亚组,共9组,每组8只.采用酶联免疫吸附法检测血清TNF-α、IL-1β,同时观察胰腺和肺组织的病理变化,免疫组织化学蛋白方法检测cAMP依赖PKA催化亚基C(PKA C)和磷酸化的血管扩张刺激磷蛋白(p-VASP),荧光定量聚合酶链反应检测肺组织VSAP mRNA表达水平.结果 与SO组比较,SAP组各时间点血清TNF-α 、IL-1β明显上升(P＜0.05),胰腺、肺病理学改变明显,肺组织PKA C蛋白、VASP磷酸化水平及VASPmRNA表达水平明显增强(P＜0.05),12 h达高峰[TNF-α(266.07±17.14) pg/mL、IL-1β (169.17±25.92) pg/mL、PKA C(210.69 ±6.32)×103、p-VASP(56.62 ±0.57)×103、VASPmRNA(2.06 ±0.21)],且与TNF-α、IL-1β之间存在明显的正相关性.与SAP组比较,SAP+ H89组各时间点胰腺、肺病理学改变明显减轻,肺组织PKA C蛋白、VASP磷酸化水平及VSAP mRNA表达水平明显下降(P＜0.05).结论 cAMP/PKA信号转导通路的活化参与了重症急性胰腺炎肺损伤的病理过程,可能与TNF-α及IL-1β水平上调及VASP磷酸化而发挥作用有关.
目的 探討環燐痠腺苷/蛋白激酶A(cAMP/PKA)信號轉導通路在重癥急性胰腺炎肺損傷的作用機製.方法 健康雄性SD大鼠72隻,按完全隨機法分為假手術(SO)組、重癥急性胰腺炎組(SAP組)、SAP+ H89(cAMP抑製劑)組,後兩組按取材時間不同又分為3、6、12及24 h四箇亞組,共9組,每組8隻.採用酶聯免疫吸附法檢測血清TNF-α、IL-1β,同時觀察胰腺和肺組織的病理變化,免疫組織化學蛋白方法檢測cAMP依賴PKA催化亞基C(PKA C)和燐痠化的血管擴張刺激燐蛋白(p-VASP),熒光定量聚閤酶鏈反應檢測肺組織VSAP mRNA錶達水平.結果 與SO組比較,SAP組各時間點血清TNF-α 、IL-1β明顯上升(P＜0.05),胰腺、肺病理學改變明顯,肺組織PKA C蛋白、VASP燐痠化水平及VASPmRNA錶達水平明顯增彊(P＜0.05),12 h達高峰[TNF-α(266.07±17.14) pg/mL、IL-1β (169.17±25.92) pg/mL、PKA C(210.69 ±6.32)×103、p-VASP(56.62 ±0.57)×103、VASPmRNA(2.06 ±0.21)],且與TNF-α、IL-1β之間存在明顯的正相關性.與SAP組比較,SAP+ H89組各時間點胰腺、肺病理學改變明顯減輕,肺組織PKA C蛋白、VASP燐痠化水平及VSAP mRNA錶達水平明顯下降(P＜0.05).結論 cAMP/PKA信號轉導通路的活化參與瞭重癥急性胰腺炎肺損傷的病理過程,可能與TNF-α及IL-1β水平上調及VASP燐痠化而髮揮作用有關.
목적 탐토배린산선감/단백격매A(cAMP/PKA)신호전도통로재중증급성이선염폐손상적작용궤제.방법 건강웅성SD대서72지,안완전수궤법분위가수술(SO)조、중증급성이선염조(SAP조)、SAP+ H89(cAMP억제제)조,후량조안취재시간불동우분위3、6、12급24 h사개아조,공9조,매조8지.채용매련면역흡부법검측혈청TNF-α、IL-1β,동시관찰이선화폐조직적병리변화,면역조직화학단백방법검측cAMP의뢰PKA최화아기C(PKA C)화린산화적혈관확장자격린단백(p-VASP),형광정량취합매련반응검측폐조직VSAP mRNA표체수평.결과 여SO조비교,SAP조각시간점혈청TNF-α 、IL-1β명현상승(P＜0.05),이선、폐병이학개변명현,폐조직PKA C단백、VASP린산화수평급VASPmRNA표체수평명현증강(P＜0.05),12 h체고봉[TNF-α(266.07±17.14) pg/mL、IL-1β (169.17±25.92) pg/mL、PKA C(210.69 ±6.32)×103、p-VASP(56.62 ±0.57)×103、VASPmRNA(2.06 ±0.21)],차여TNF-α、IL-1β지간존재명현적정상관성.여SAP조비교,SAP+ H89조각시간점이선、폐병이학개변명현감경,폐조직PKA C단백、VASP린산화수평급VSAP mRNA표체수평명현하강(P＜0.05).결론 cAMP/PKA신호전도통로적활화삼여료중증급성이선염폐손상적병리과정,가능여TNF-α급IL-1β수평상조급VASP린산화이발휘작용유관.
Objective To investigate the mechanism of cyclic adenosine monophosphate / protein kinase A signal transduction pathway in severe acute pancreatitis(SAP)-associated lung injury.Methods Seventy-two male healthy SD rats were completely randomized into three groups:sham operation (SO) group(n =8),SAP group and SAP plus H89 (cAMP inhibitor) group,then the latter two groups were divided into four sub-groups with eight rats in each sub-group according to the sampling time of 3,6,12 and 24 h,and the total numbers of groups were nine.The content change of TNF-α and IL-1β in serum,protein levels of cAMP-dependent protein kinase catalytic subunit (PKA C) and phosphorylated vasodilator-stimulated phosphoprotein(p-VASP) and the expression of VSAP mRNA in lung tissue were detected by enzyme-linked immunosorbent assay (ELISA),immunohistochemistry and quantitative real time PCR,respectively.Pathological changes of the pancreas and lung tissues were also observed.Results Compared with the SO group,the serum levels of TNF-α and IL-1β in the SAP group were obviously increased at different time points(P ＜0.05).Pathological changes of the pancreas and lung tissues were aggravated significantly.The protein levels of PKA C,p-VASP and the expression of VSAP mRNA in lung tissue were increased significantly (P ＜0.05)which peaked at 12 h in the SAP group [TNF-α was (266.07 ± 17.14) pg/mL,IL-1β(169.17 ±25.92) pg/mL,PKA C(210.69 ±6.32) × 103,p-VASP (56.62 ±0.57) × 103,VASP mRNA(2.06 ±0.21)],which had positive correlation with the serum level of TNF-α and IL-1β.Compared with the SAP group,pathological changes of the pancreas and lung tissues were alleviated significantly,the protein levels of PKA C,p-VASP and the expression of VSAP mRNA in lung tissue were decreased significantly in the SAP plus H89 group at different time points(P ＜ 0.05).Conclusion The cyclic adenosine monophosphate / protein kinase A signal transduction pathway is found to participate in the pathological process of SAP-associated lung injury through the up-regulations of TNF-α,IL-1 β and phospho-VASP.
