摘要
目的探讨ATP敏感性钾通道（KATP）在-氧化氮（NO）调节失血性休克（HS）大鼠离体淋巴管收缩性中的作用机制。方法84只雄性Wistar大鼠按随机数字表法分为对照组（n=6）、HS0．5h组（n=36）、HS2h组（n=42）；制备离体淋巴管条，分别或联合与K。阻断剂格列本脲（Gli）及其开放剂吡那地尔（Pin）、NO供体L-精氨酸（L—Arg）、蛋白激酶A（PKA）抑制剂H-89及其供体8-溴-环磷酸腺苷（8-Br—cAMP）、-氧化氮合酶（NOS）抑制剂L-硝基-精氨酸甲脂（L—NAME）、可溶性鸟苷酸环化酶抑制剂1h-[1，2，4]藤二唑[4，3-a]喹喔啉-1-酮（ODQ）、蛋白激酶G（PKG）抑制剂KT-5823共同孵育（分别为HS0．5h、HS0．5h＋L—Arg、HS0．5h＋L—Arg＋H-89、HS0．5h＋I，-Arg＋Gli、HS0．5h＋8-Br-cAMP、HS0．5h＋8-Br—cAMP＋Gli组及HS2h、HS2h十L—NAME、HS2h＋L—NAME手Pin，HS2h十ODQ？HS2h手ODQ手Pin、HS2h手KT-5823、HS2h手KT-5823＋Pin组，月=6）。采用离体淋巴管灌流技术，记录淋巴管收缩频率（CF），计算收缩幅度（CA）、紧张指数（TI）和泵流分数（FPF）。结果HS0．5h淋巴管的CF、TI、FPF显著高于对照组；L—Arg能显著降低HS0．5h淋巴管的cF、TI、FPF，H-89、Gli分别与L—Arg共孵育后，H-89能够逆转L—Arg降低CF、FPF的作用，Gli能够逆转L—Arg降低FPF的作用；8-Br—cAMP能降低HS0．5h淋巴管的CF、FPF，Gli与8-Br—cAMP共孵育后，CF显著高于HS0．5h＋8-Br—cAMP组，且TI、FPF显著低于HS0．5h组。HS2h淋巴管的CF、FPF、TI较对照组显著降低；L—NAME可提高HS2h淋巴管的cF、TI、FPF，ODQ可提高HS2h淋巴管的CF、TI，KT-5823可提高Hs2h淋巴管的CF、FPF；而L-NAME、ODQ、KT－5823分别与Pin共孵育后，分别能显著降低HS2h淋巴管单独与L—NAME孵育时的CF、FPF，降低与单独ODQ孵育时的CF、TI、FPF，并恢复CA至对照组水平，降低单独与KT-5823孵育时的CF、FPF。结论KATP参与了NO对HS大鼠离体淋巴管泵功能的调节作用，其作用与NO—cAMP—PKA、NO—cGMP—PKG信号通路有关。
Objective To investigate the mechanism of ATP-sensitive potassium channel （KAy） on nitride oxide （NO） regulating contractile activity of isolated lymphatics from hemorrhagic shock （HS） rats. Methods Eighty - four Wistar rats were randomly divided into control group （n=6）, HS 0.5-hour group （n=36）, HS 2-hour group （n：42）. A segment of thoracic duct of rats was adopted for isolated lymphatics after HS, then the HS 0.5-hour and 2-hour lymphatics were incubated combinedly or respectively with KATP inhibitor glibenclaimde （Gli）, opener of KATp pinacidil （Pin）, NO doner L-arginine （L-Arg）, protein kinase A （PKA） inhibitor H-89, PKA doner 8-bromine-cyclic adenosine monophosphate （8-Br-cAMP）, nitricoxide synthase （NOS） inhibitor N-nitro-L-arginine methyl ester （L-NAME）, soluble guanylyl cyclase （sGC） inhibitor 1 h-[1,2,4]-oxadiazole-[4,3-a]- quinoxalin-l-one （ODQ）, protein kinase G （PKG） inhibitor KT-5823 （named as HS 0.5 h, HS 0.5 h ＋ L-Arg, HS 0.5 h ＋ L-Arg ＋ H-89, HS 0.5 h ＋ L-Arg ＋ Gli, HS 0.5 h ＋ 8-Br-cAMP, HS 0.5 h ＋ 8-Br-cAMP ＋ Gli and HS 2 h, HS 2 h ＋ L-NAME, HS 2 h ＋ L-NAME ＋ Pin, HS 2 h ＋ ODQ, HS 2 h ＋ ODQ ＋ Pin, HS 2 h ＋ KT-5823, HS 2 h ＋ KT-5823 ＋ Pin, n=6）. By lymphatic perfusion in vitro, contractile frequency （CF） was recorded, and contractile amplitude （CA）, tonic index （TI） and fractional pump flow （FPF） were calculated. Results The results suggested that the CF, TI, FPF of HS 0.5-hour lymphatics were significantly increased compared with control group, and the CF, TI, FPF decreased significantly when incubated with L-Arg. H-89 could deteriorate the decreased effect of L-Arg on CF and FPF, and Gli could deteriorate the decreased effect of L-Arg on FPF. When the HS 0.5-hour lymphatics incubated with 8-Br-cAMP, the CF and FPF were all decreased significantly, and when the HS 0.5-hour lymphatics incubated with 8-Br-cAMP and Gli, the CF was significantly higher than HS 0.5 h ＋ 8-Br-cAMP group, and the TI and FPF decreased significantly compared with HS 0.5-hour group. The CF, FPF, TI of HS 2-hour lymphatics were significantly decreased compared with control group. L-NAME could increase the CF, TI, FPF ; ODQ could increase the CF, TI; KT-5823 could increase the CF and FPF; when incubated with Pin respectively, the CF and FPF when incubated with L-NAME were decreased, the CF, TI and FPF when incubated with ODQ were decreased, in addition, the CA was recovered as level of control group, the CF and FPF when incubated with KT-5823 were decreased. Conclusion KATP involved in NO modulating pumping function of isolated lymphatics of HS rats, and the effect may be relative to the signal pathway of NO-cAMP-PKA and NO-cGMP-PKG.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2012年第8期457-460,共4页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目（30770845）
河北省自然科学基金资助项目（C2008000503）
河北省教育厅科学研究重点项目（ZH2007101,ZD20100201）
