中国危重病急救医学
中國危重病急救醫學
중국위중병급구의학
CHINESE CRITICAL CARE MEDICINE
2012年
8期
457-460
,共4页
张立民%牛春雨%赵自刚%司永华%张玉平
張立民%牛春雨%趙自剛%司永華%張玉平
장립민%우춘우%조자강%사영화%장옥평
失血性休克%离体实验%淋巴管%收缩性%一氧化氮%ATP敏感性钾通道
失血性休剋%離體實驗%淋巴管%收縮性%一氧化氮%ATP敏感性鉀通道
실혈성휴극%리체실험%림파관%수축성%일양화담%ATP민감성갑통도
Hemorrhagic shock%In vitro%Lymphatics%Contractility%Nitric oxide%ATP-sensitive potassium channel
目的 探讨ATP敏感性钾通道(KATP)在一氧化氮(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)、HS 0.5 h组(n=36)、HS2h组(n=42)；制备离体淋巴管条,分别或联合与KATP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)、一氧化氮合酶(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1 h-[1,2,4](恶)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HS 0.5 h、HS 0.5 h+L-Arg、HS0.5 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组及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、HS2h+KT-5823+ Pin组,n=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 HS 0.5 h淋巴管的CF、TI、FPF显著高于对照组；L-Arg能显著降低HS 0.5 h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HS 0.5 h淋巴管的CF、FPF,Gli与8-Br-cAMP共孵育后,CF显著高于HS 0.5 h+8-Br-cAMP组,且TI、FPF显著低于HS 0.5 h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.
目的 探討ATP敏感性鉀通道(KATP)在一氧化氮(NO)調節失血性休剋(HS)大鼠離體淋巴管收縮性中的作用機製.方法 84隻雄性Wistar大鼠按隨機數字錶法分為對照組(n=6)、HS 0.5 h組(n=36)、HS2h組(n=42)；製備離體淋巴管條,分彆或聯閤與KATP阻斷劑格列本脲(Gli)及其開放劑吡那地爾(Pin)、NO供體L-精氨痠(L-Arg)、蛋白激酶A(PKA)抑製劑H-89及其供體8-溴-環燐痠腺苷(8-Br-cAMP)、一氧化氮閤酶(NOS)抑製劑L-硝基-精氨痠甲脂(L-NAME)、可溶性鳥苷痠環化酶抑製劑1 h-[1,2,4](噁)二唑[4,3-a]喹喔啉-1-酮(ODQ)、蛋白激酶G(PKG)抑製劑KT-5823共同孵育(分彆為HS 0.5 h、HS 0.5 h+L-Arg、HS0.5 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組及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、HS2h+KT-5823+ Pin組,n=6).採用離體淋巴管灌流技術,記錄淋巴管收縮頻率(CF),計算收縮幅度(CA)、緊張指數(TI)和泵流分數(FPF).結果 HS 0.5 h淋巴管的CF、TI、FPF顯著高于對照組；L-Arg能顯著降低HS 0.5 h淋巴管的CF、TI 、FPF,H-89、Gli分彆與L-Arg共孵育後,H-89能夠逆轉L-Arg降低CF、FPF的作用,Gli能夠逆轉L-Arg降低FPF的作用；8-Br-cAMP能降低HS 0.5 h淋巴管的CF、FPF,Gli與8-Br-cAMP共孵育後,CF顯著高于HS 0.5 h+8-Br-cAMP組,且TI、FPF顯著低于HS 0.5 h組.HS2h淋巴管的CF、FPF、TI較對照組顯著降低；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分彆與Pin共孵育後,分彆能顯著降低HS2h淋巴管單獨與L-NAME孵育時的CF、FPF,降低與單獨ODQ孵育時的CF、TI、FPF,併恢複CA至對照組水平,降低單獨與KT-5823孵育時的CF、FPF.結論 KATP參與瞭NO對HS大鼠離體淋巴管泵功能的調節作用,其作用與NO-cAMP-PKA、NO-cGMP-PKG信號通路有關.
목적 탐토ATP민감성갑통도(KATP)재일양화담(NO)조절실혈성휴극(HS)대서리체림파관수축성중적작용궤제.방법 84지웅성Wistar대서안수궤수자표법분위대조조(n=6)、HS 0.5 h조(n=36)、HS2h조(n=42)；제비리체림파관조,분별혹연합여KATP조단제격렬본뇨(Gli)급기개방제필나지이(Pin)、NO공체L-정안산(L-Arg)、단백격매A(PKA)억제제H-89급기공체8-추-배린산선감(8-Br-cAMP)、일양화담합매(NOS)억제제L-초기-정안산갑지(L-NAME)、가용성조감산배화매억제제1 h-[1,2,4](악)이서[4,3-a]규악람-1-동(ODQ)、단백격매G(PKG)억제제KT-5823공동부육(분별위HS 0.5 h、HS 0.5 h+L-Arg、HS0.5 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+ Gli조급HS 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、HS2h+KT-5823+ Pin조,n=6).채용리체림파관관류기술,기록림파관수축빈솔(CF),계산수축폭도(CA)、긴장지수(TI)화빙류분수(FPF).결과 HS 0.5 h림파관적CF、TI、FPF현저고우대조조；L-Arg능현저강저HS 0.5 h림파관적CF、TI 、FPF,H-89、Gli분별여L-Arg공부육후,H-89능구역전L-Arg강저CF、FPF적작용,Gli능구역전L-Arg강저FPF적작용；8-Br-cAMP능강저HS 0.5 h림파관적CF、FPF,Gli여8-Br-cAMP공부육후,CF현저고우HS 0.5 h+8-Br-cAMP조,차TI、FPF현저저우HS 0.5 h조.HS2h림파관적CF、FPF、TI교대조조현저강저；L-NAME가제고HS2h림파관적CF、TI、FPF,ODQ가제고HS2h림파관적CF、TI,KT-5823가제고HS2h림파관적CF、FPF;이L-NAME、ODQ、KT-5823분별여Pin공부육후,분별능현저강저HS2h림파관단독여L-NAME부육시적CF、FPF,강저여단독ODQ부육시적CF、TI、FPF,병회복CA지대조조수평,강저단독여KT-5823부육시적CF、FPF.결론 KATP삼여료NO대HS대서리체림파관빙공능적조절작용,기작용여NO-cAMP-PKA、NO-cGMP-PKG신호통로유관.
Objective To investigate the mechanism of ATP-sensitive potassium channel (KATP) 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 berelative to the signal pathway of NO-cAMP-PKA and NO-cGMP-PKG.
