【作者】 林剑清；

【导师】 陈国忠；

【作者基本信息】 福建医科大学 ， 麻醉学， 2008， 硕士

【摘要】 目的:建立简单、重复率高的兔的SAH后的CVS模型和HTEB模型,证实HTEB对SAH后的CVS形成的预防作用,并进一步探讨HTEB预防SAH后的CVS形成的可能机制。方法:1.采用切开直接置入硬膜外导管法建立兔上胸段硬膜外阻滞的模型。2.硬膜外阻滞成功的新西兰兔60只,随机分为6组（n=10）:正常1 d组（N1组）、正常7 d组（N7组）、单纯SAH 1 d组（S1组）、单纯SAH 7 d组（S7组）、HTEB+SAH 1 d组（HS1组）、HTEB+SAH 7 d组（HS7组）;N1、N7组硬膜外导管推注生理盐水（NS）1 mL后接镇痛泵持续泵入NS 1 mL/h,30 min后枕大池推注NS 0.5 mL/kg;S1、S7组硬膜外推注NS 1 mL后接镇痛泵泵入NS 1 mL/h,30 min后制备SAH模型;HS1、HS7组硬膜外推注1 g·L^-1罗哌卡因1 mL后接镇痛泵泵入1 g·L^-1罗哌卡因1 mL/h,30 min后制备SAH模型。3.采用经皮枕大池穿刺自体动脉血注入法建立兔的SAH后的CVS模型。4.观察动物的进食量比较N7、HS7与S7三组兔日食量的差别。5.用经颅多普勒（TCD）动态检测基底动脉血流动力学指标,反映血管痉挛的情况。6.开颅取脑后,先进行大体标本观察;再剪取基底动脉下1/3,放入标本杯,40 g·L^-1多聚甲醛浸泡固定,备石蜡包埋切片、HE染色,观察基底动脉。剪取基底动脉上2/3,放入Eppendof管,放入液氮保存,备RT-PCR检测基底动脉preproET-lmRNA和eN0SmRNA表达含量变化。结果:1.食量:N7组1 d减少（P<0.05）,2 d后无明显变化（P>0.05）;S7组1～3 d（P<0.01）和4～7 d（P <0.05）均明显减少;HS7组1～2 d明显减少（P<0.01）,4 d后明显恢复（P>0.05）。2.大体标本可见S1组、S7组、HS1组、HS7组脑底面可见血凝块,以基底动脉周围最多,脑底池呈暗红色,N1组、N7组基底动脉清晰可见,无血凝块形成。3.兔基底动脉N1组、S1组、HS1组、N7组、S7组、HS7组的TCD指标变化。Vs和Vm: S1组较N1组明显加快（P<0.01）,S7组较N7组明显加快（P<0.01）,HS1组较N1组有加快（P<0.05）,HS7组和N7组相比差异不明显（P>0.05）。PI和RI:S1组较N1组明显降低（P<0.01）,S7组较N7组明显降低（P<0.01）。4. BA光镜观察:N1、N7组未见异常,S1、S7组管壁收缩、管腔狭窄,HS1、HS7组变化程度均轻于S1、S7组,HS7组狭窄程度轻于HS1。5. preproET-lmRNA的表达:与N1组相比,S1组表达显著增多（P?0.01）,HS1组也增多（P?0.05）;与N7组相比,S7组显著增多（P?0.01）,HS7组也增多（P?0.05）;与HS1相比,HS7组表达降低（P?0.05）。eN0SmRNA的表达:与N1组相比,S1组表达显著降低（P?0.01）,HS1组也降低（P?0.05）;与N7组相比,S7组显著降低（P?0.01）,HS7组也降低（P?0.05）;与HS1相比,HS7组表达增多（P?0.05）。结论:1.采用经T6-7切开置管法可建立兔上胸段硬膜外阻滞的模型。2.采用经皮枕大池穿刺自体动脉血注入法可建立兔的SAH后的CVS模型。3.预防性应用HTEB可以减轻SAH后CVS的程度,治疗时间越长效果越好。4.预防性应用HTEB可以下调SAH后基底动脉痉挛的内皮细胞的preproET-lmRNA的表达水平和上调eN0SmRNA的表达水平。更多还原

【Abstract】 Objective:The purpose of the study was to establish two simple and inexpensive animal models of experimental CVS after SAH and HTEB in rabbits and to investigate the preventive effects of HTEB on CVS following SAH and to study the possible mechanism of the preventive effects .Methods:1. Sixty rabbits with HTEB made successfully were randomly assigned into 6 groups （n = 10,each）: the normal 1 day group （N1）、the normal 7 day group（N7）、SAH 1 day group （S1）、SAH 7 day group （S7）、HTEB plus SAH 1 day group （HS1）、HTEB plus SAH 7 day group （HS7）. the normal group （N1、N7）: normal saline 0.5 mL/kg was injected into the cisterna magna and 1 mL/h injected through the epidural pipe; SAH group （S1、S7）: non-anticoagulant autologous arterial blood 0.5 mL/kg was injected into the cisterna magna and normal saline 1mL/h was injected through the epidural pipe; HTEB plus SAH group （HS1、HS7）: non-anticoagulant autologous arterial blood 0.5 mL/kg was injected into the cisterna magna and 1 g·L^-1 Ropivacaine 1 mL/h injected through the epidural pipe.2. To establish the HTEB model, T6-7 epidural space was explored, and an epidural pipe was introduced. The 1 g·L^-1 ropivacaine was injected continually through the pipe at 1 mL/h to block T1-5 sympathetic nerves.3. SAH models of rabbits were developed by injecting non-anticoagulant autologous arterial blood 0.5 mL/kg into the cisterna magna.4. In-taking food of group N7、S7、HS7 was evaluated every day for 7 days..5. Transcranial Doppler （TCD） ultrasonography was used to detect the basilar artery blood flow velocity changes.6. After taking out the brain by opening skulls, a general observation of specimens was made firstly. Then one-third of basilar arteries were preservinged and fixed by 40 g·L^-1 paraformaldehyde, and paraffin-embedded、sected、HE stained. Lastly the basilar artery changes were observed through Optical Microscope. Two-third of basilar arteries were preserved in liquid nitrogen by Eppendof tubes and was used to detect the expressions of preproET-lmRNA and eN0SmRNA by RT-PCR.Results:1. In-taking food was not significant decreased in group N7 except the first day after injecting Sodium Chloride into the cisterna magna. In group S7, it was decreased significantly through the 7 days observative period. It was decreased in the first two days, and returned to normal from the fourth day in group HS7.2. The changes of pathology: By eyes there were blood clots scattering on the basal surface of the brain and around the basilar artery in group HS1、HS7 and S1、S7. But there were no blood clots in group N1、N7.3. Changes of vascular hemodynamic indexes were showed by TCD. Vs and Vm in group S1 and S7 were increased significantly compared with group N1 and N7 （P<0.01）; but PI and RI were decreased respectively （P<0.05）. Vm and Vs in group HS1 and HS7 were decreased compared with group S1 and S7（P<0.05）, but PI and RI were increased respectively（P<0.05）. 4. Pathological observation:The structure of the basilar artery in group N1 and N7 were normal. The vessel wall was shrink and the lumina became narrow in group S1 and S7. The pathologic changes in group HS1 and HS7 were slighter than in group S1 and S7.5. The expression of preproET-lmRNA of group S1 and S7 were higher than group N1 and N7（P?0.05）,and group HS1 and HS7 higher than group N1 and N7（P?0.05）.The expression of eN0SmRNA of group S1 and S7 were decreased significantly compared with N1 and N7 respectively（P?0.05）;but there were no significant differences between group N7 and HS7 （P>0.05）.Conclusions:1. The HTEB rabbit model can be established successfully by incision T6-7 epidural space and introducing epidural pipe.2. the CVS following SAH rabbit model can be established successfully by injecting non-anticoagulant autologous arterial blood 0.5mL/kg into the cisterna magna.3. HTEB used before SAH can reduce the extent of CVS following SAH, and the longer of the treatment, the better of the effect.4. HTEB used before SAH can inhibit the expressions of preproET-lmRNA and promote the expressions of eNOSmRNA of the vasospasm BA.更多还原