枕大池注入利多卡因对兔SAH后基底动脉Rho/ROCK信号传导的影响
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摘要
目的探讨枕大池注入利多卡因对兔蛛网膜下腔出血(SAH)后基底动脉血管Rho/Rho相关激酶(ROCK)信号传导的影响。方法 24只新西兰大白兔随机分为3组,每组8只,分别为假手术(sham)组、蛛网膜下腔出血(SAH)组、枕大池利多卡因(CD)组。SAH组和CD组动物经枕大池注入非抗凝的自体动脉血(1mL/kg)复制SAH模型,sham组经枕大池注入37℃的生理盐水(1mL/kg);30min后,CD组经枕大池注入0.3mL 2%利多卡因,SAH组和sham组注入等量生理盐水。72h后测定摄食量和神经功能损害分级,实时荧光定量PCR(qRT-PCR)和Western blot检测基底动脉中Rho相关激酶2(ROCK2)、肌球蛋白轻链(MLC)及钙调蛋白(CaM)蛋的基因和蛋白表达。结果与sham组比较,SAH组和CD组家兔摄食量减少,且出现不同程度的神经功能损害,ROCK2在基底动脉中的mRNA及蛋白表达量增高,MLC和CaM的mRNA和蛋白表达减少(P<0.05);与SAH组比较,CD组家兔摄食量及神经功能损害差异无统计学意义(P>0.05),ROCK2在基底动脉中的mRNA及蛋白表达量减少,MLC和CaM的mRNA和蛋白表达增高(P<0.05)。结论枕大池注入利多卡因能够抑制兔SAH后基底动脉血管Rho/ROCK信号传导,减轻SAH后基底动脉平滑肌的收缩。
Objective To observe the expression of Rho/Rho-associated kinase(ROCK)signaling pathway in the basilar artery and the effect of lidocaine on this signaling pathway after subarachnoid hemorrhage(SAH)in rabbits.Methods 24 New Zealand white rabbits were randomly divided into sham operation(sham)group,SAH group,and occipital cisterna lidocaine(CD)group.There were 8 rabbits in each group.Intracisternal injection of non-anticoagulant autologous arterial blood(1 mL/kg)were applied to SAH group and CD group animals to establish SAH model,sham normal saline group was injected with 37 ℃ physiological saline(1 mL/kg);after30 min,CD group was injected with 0.3 mL 2%lidocaine cisterna,SAH group and sham group were injected with saline.After 72 h,food intake and neurologic function damage were measured.The expressions of Rho associated kinase 2(ROCK2)and myosin light chain(MLC)and calmodulin(CaM)protein in the basilar artery were measured by Western blot.The ROCK2 and MLCand CaM gene expressions were measured by using real-time quantitative PCR.Results Compared with sham group,reduced food intake,various degrees of neurological impairment,increased ROCK2 mRNA and protein expressions in basal artery,and decreased MLC and CaM expressions were observed in SAH group and CD group(P<0.05).Compared with the SAH group,there was no statistically significant difference in diet intake and neurological damage in the CD group(P>0.05);the mRNA and protein expressions of ROCK2 in the basilar artery decreased,and the expressions of MLC and CaM increased(P<0.05).Conclusion Intracisternal injection of lidocaine may inhibit the transmission of Rho/ROCK signal in the basilar artery and reduce the basilar artery smooth muscle contraction after SAH.
Objective To observe the expression of Rho/Rho-associated kinase(ROCK)signaling pathway in the basilar artery and the effect of lidocaine on this signaling pathway after subarachnoid hemorrhage(SAH)in rabbits.Methods 24 New Zealand white rabbits were randomly divided into sham operation(sham)group,SAH group,and occipital cisterna lidocaine(CD)group.There were 8 rabbits in each group.Intracisternal injection of non-anticoagulant autologous arterial blood(1 mL/kg)were applied to SAH group and CD group animals to establish SAH model,sham normal saline group was injected with 37 ℃ physiological saline(1 mL/kg);after30 min,CD group was injected with 0.3 mL 2%lidocaine cisterna,SAH group and sham group were injected with saline.After 72 h,food intake and neurologic function damage were measured.The expressions of Rho associated kinase 2(ROCK2)and myosin light chain(MLC)and calmodulin(CaM)protein in the basilar artery were measured by Western blot.The ROCK2 and MLCand CaM gene expressions were measured by using real-time quantitative PCR.Results Compared with sham group,reduced food intake,various degrees of neurological impairment,increased ROCK2 mRNA and protein expressions in basal artery,and decreased MLC and CaM expressions were observed in SAH group and CD group(P<0.05).Compared with the SAH group,there was no statistically significant difference in diet intake and neurological damage in the CD group(P>0.05);the mRNA and protein expressions of ROCK2 in the basilar artery decreased,and the expressions of MLC and CaM increased(P<0.05).Conclusion Intracisternal injection of lidocaine may inhibit the transmission of Rho/ROCK signal in the basilar artery and reduce the basilar artery smooth muscle contraction after SAH.
引文
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[4]吴科,魏剑波,张月辉,等.自发性蛛网膜下腔出血后脑血管痉挛节律特征的研究.四川生理科学杂志,2012,34(4):155-156.
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[7]PUETZ S,LUBOMIROV LT,PFITZER G.Regulation of smooth muscle contraction by small GTPases.Physiology(Bethesda),2009,24(6):342-356.
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[9]RODRIGUEZ A,EGEA-GUERRERO J,RUIZ D,et al.Biomarkers of vasospasm development and outcome in aneurysmal subarachnoid hemorrhage.J Neurol Sci,2014,341(1/2):119-127.
[10]DANURA H,SCHATLO B,MARBACHER S,et al.Acute angiographic vasospasm and the incidence of delayed cerebral vasospasm preliminary results.Acta Neurochir Suppl,2015,120(120):187-190.
[11]国伟,孟建中,陈宇.Rho/Rho激酶信号通路与血管内皮通透性的研究.生物医学工程研究,2009,28(2):154-158.
[12]AMANO M,NAKAYAMA M,KAIBUCHI K.Rhokinase/ROCK:a key regulator of the cytoskeleton and cell polarity.Cytoskeleton,2010,67(9):545-554.
[13]MORGAN-FISHER M,WEWER UM,YONEDA A.Regulation of ROCK activity in cancer.J Histochem Cytochem,2013,61(3):185-198.
[14]PARK J,KIM S,KIM H,et al.Transretroperitoneal CTguided embolization of growing internal iliac artery aneurysm after repair of abdominal aortic aneurysm:a trasretroperitoneal approach with intramuscular lidocaine injection technique.Cardiovasc Intervent Radiol,2015,38(1):227-231.
[15]LIN C,TSAI Y,WANG H,et al.Pre-emptive trement of lidocaine attenuates neuropathic pain and reduces pain-related biochemical markers in the rat cuneate necleus in median nerve chronic constriction injury model.Anesthesiol Res Pract,2012,20(12):921405.
[16]SHIRAO S,FUJISAWA H,KUDO A,et al.Inhibitory Effects of eicosapentaenoic acid on chronic cerebral vasospasm after subarachnoid hemorrhage:possible involvement of a sphingosylphosphorylcholine-rho-kinase pathway.Cerebrovasc Dis,2008,26(1):30-37.
[17]YUAN TY,YAN Y,WU YJ,et al.Vasodilatory effect of a novel Rho-kinase inhibitor,DL0805-2,on the rat mesenteric artery and its potential mechanisms.Cardiovasc Drugs Ther,2014,28(5):415-424.
[18]赵炜疆.Rho激酶与脑血管痉挛综述.基础医学与临床,2001,25(10):896-990.
[19]魏华.利多卡因治疗椎基底动脉供血不足35例的临床观察.中外医学研究,2015,31(11):145-147.
[2]D’SOUZA S.Aneurysmal subarachnoid hemorrhage.Neurosurg Anesthesiol,2015,27(3):222-240.
[3]FU M,YAN J,ROLLAND WB,et al.Early brain injury,an evolving frontier in subarachnoid hemorrhage research.Transl Stroke Res,2013,4(4):432-446.
[4]吴科,魏剑波,张月辉,等.自发性蛛网膜下腔出血后脑血管痉挛节律特征的研究.四川生理科学杂志,2012,34(4):155-156.
[5]PRZYBCIEN-SZYMANSKA MM,ASHLEY WW.Biomarker discovery in cerebral vasospasm after aneurysmal subarachnoid hemorrhage.Stroke Cerebrovasc Dis,2015,24(7):1453-1464.
[6]LOIRAND G,PACAUD P.The role of Rho protein signaling in hypertension.Nat Rev Cardiol,2010,7(11):637-647.
[7]PUETZ S,LUBOMIROV LT,PFITZER G.Regulation of smooth muscle contraction by small GTPases.Physiology(Bethesda),2009,24(6):342-356.
[8]ENDO S,BRANSON PJ,ALKSNE JF.Experimental model of sumptomatic vasospasmin rabbits.Ssroke,1988,19(11):1420-1425.
[9]RODRIGUEZ A,EGEA-GUERRERO J,RUIZ D,et al.Biomarkers of vasospasm development and outcome in aneurysmal subarachnoid hemorrhage.J Neurol Sci,2014,341(1/2):119-127.
[10]DANURA H,SCHATLO B,MARBACHER S,et al.Acute angiographic vasospasm and the incidence of delayed cerebral vasospasm preliminary results.Acta Neurochir Suppl,2015,120(120):187-190.
[11]国伟,孟建中,陈宇.Rho/Rho激酶信号通路与血管内皮通透性的研究.生物医学工程研究,2009,28(2):154-158.
[12]AMANO M,NAKAYAMA M,KAIBUCHI K.Rhokinase/ROCK:a key regulator of the cytoskeleton and cell polarity.Cytoskeleton,2010,67(9):545-554.
[13]MORGAN-FISHER M,WEWER UM,YONEDA A.Regulation of ROCK activity in cancer.J Histochem Cytochem,2013,61(3):185-198.
[14]PARK J,KIM S,KIM H,et al.Transretroperitoneal CTguided embolization of growing internal iliac artery aneurysm after repair of abdominal aortic aneurysm:a trasretroperitoneal approach with intramuscular lidocaine injection technique.Cardiovasc Intervent Radiol,2015,38(1):227-231.
[15]LIN C,TSAI Y,WANG H,et al.Pre-emptive trement of lidocaine attenuates neuropathic pain and reduces pain-related biochemical markers in the rat cuneate necleus in median nerve chronic constriction injury model.Anesthesiol Res Pract,2012,20(12):921405.
[16]SHIRAO S,FUJISAWA H,KUDO A,et al.Inhibitory Effects of eicosapentaenoic acid on chronic cerebral vasospasm after subarachnoid hemorrhage:possible involvement of a sphingosylphosphorylcholine-rho-kinase pathway.Cerebrovasc Dis,2008,26(1):30-37.
[17]YUAN TY,YAN Y,WU YJ,et al.Vasodilatory effect of a novel Rho-kinase inhibitor,DL0805-2,on the rat mesenteric artery and its potential mechanisms.Cardiovasc Drugs Ther,2014,28(5):415-424.
[18]赵炜疆.Rho激酶与脑血管痉挛综述.基础医学与临床,2001,25(10):896-990.
[19]魏华.利多卡因治疗椎基底动脉供血不足35例的临床观察.中外医学研究,2015,31(11):145-147.