大鼠静脉局部麻醉模型的建立与阿米替林静脉局部麻醉作用的研究

发布时间：2018-06-06 12:58

本文选题：静脉局部麻醉 + 大鼠　；参考：《四川大学》2007年硕士论文

【摘要】： 背景：静脉局部麻醉(Intravenous regional anesthesia；IVRA)，又称Bier Block，于1908年开始应用于临床。此种方法主要适用于成人的四肢短小手术，具有经济、简便、效果确切、无神经创伤等优点，现欧美等国家仍广泛应用。目前报道应用于IVRA研究的动物模型有狗和兔子等，但此类模型价格昂贵、操作复杂、重复性差，因此需要建立新的经济实用的动物模型。阿米替林是一种传统的三环类抗抑郁药，主要经全身用药治疗抑郁症、缓解神经病理性疼痛等。最近动物实验研究发现阿米替林可以产生与传统局麻药类似的神经阻滞作用。本研究旨在建立大鼠IVRA模型，利用该模型研究阿米替林的IVRA效能、作用时间、局部应用毒性、安全范围等，探讨阿米替林在IVRA中应用的可能性，为临床应用提供理论及实验基础。第一部分大鼠IVRA模型的建立目的： (1)建立大鼠IVRA模型(2)验证大鼠IVRA模型中药物扩散(3)观察模型中止血带阻断时间对鼠尾神经功能影响。方法： (1)健康成年雄性SD大鼠16只，随机均分为0.5％利多卡因实验组和生理盐水对照组。将鼠尾均分成三段，即上段、中段、末段。进行鼠尾静脉穿刺、驱血、固定止血带(鼠尾上段、中段交界处)、注药等操作建立大鼠IVRA模型。以夹尾反应阴性(夹尾无逃避等反应)做为鼠尾局部麻醉作用的有效指标，以甩尾反应时间(Tail-Flick Latency，TFL)恢复至1—4秒基础范围内为鼠尾感觉神经恢复的判断指标。观察并记录给药后鼠尾各段夹尾反应、TFL以及松止血带后夹尾反应、甩尾反应的恢复时间，并于第二日观察TFL及鼠尾皮肤情况。 (2)健康成年雄性大鼠3只，如实验第(1)建立大鼠IVRA模型，给予美蓝0.5ml。通过观察给药后鼠尾皮肤颜色变化，验证模型中药物的扩散。 (3)健康成年雄性大鼠24只，随机均分为四组：止血带阻断时间为10分钟组(S_(10)组)，15分钟组(S_(15)组)，30分钟组(S_(30)组)，60分钟组(S_(60)组)。如实验(1)建立大鼠IVRA模型，给予生理盐水0.5ml，四组分别留置不同时间的止血带。观察给予盐水后及松止血带前1分钟夹尾反应，松止血带后第1、5、10、20、30、60分钟、第二日的TFL及第二日的鼠尾皮肤情况。结果： (1)利多卡因组：大鼠给药后仅产生鼠尾中、末段(止血带以下)的夹尾反应阴性，且TFL为10秒。松止血带后鼠尾中段、末段夹尾反应恢复时间分别为17.5±11.2分钟，26.4±19.6分钟，；TFL恢复时间分别为30.3±20.5分钟，41.5±29.4分钟。鼠尾中、末段恢复时间比较均有统计学差异(P＜0.05)。生理盐水组：大鼠给药后鼠尾中、末段夹尾反应阳性，各时点TFL均＜4秒，TFL和基础值无统计学差异(P＞0.05)。两组鼠尾上段夹尾反应阳性，各时点TFL均＜4秒，和基础值比较无统计学差异(P＞0.05)。两组大鼠第二日鼠尾各段TFL和基础值比较均无统计学差异，鼠尾皮肤完好。 (2)3只大鼠给药后仅止血带以下的鼠尾皮肤变为蓝黑色。 (3)四个不同止血带时间组，大鼠给予生理盐水后夹尾反应均为阳性；松止血带后仅S_(60)组鼠尾中、末段第1分钟TFL＞4秒，和基础值比较有统计学差异(P＜0.05)。其余各组各时间点TFL和基础值比较均无统计学差异(P＞0.05)。大鼠第二日各组鼠尾各段TFL和基础值比较均无统计学差异，鼠尾皮肤完好。结论：大鼠IVRA简单易行，经济实用，成功率高，具有临床IVRA特点，可以用作IVRA的药物相关研究等。第二部分阿米替林、布比卡因及利多卡因IVRA效能的比较目的：本实验采用序贯法研究阿米替林IVRA作用的半数有效浓度(EC_(50))，并和传统的局麻药布比卡因、利多卡因比较，探讨阿米替林的IVRA麻醉效能。方法：体重为190—240克的健康成年雄性大鼠90只，随机均分为阿米替林组、布比卡因组及利多卡因组。建立大鼠IVRA模型，以鼠尾中段夹尾反应阴性作为鼠尾局部麻醉作用的判断指标，以序贯法分别求出三种药物的EC_(50)值，直到实验中出现有效到无效6个交叉点时终止实验。计算EC_(50)值及药物效价比。结果：阿米替林、布比卡因及利多卡因的IVRA作用EC_(50)分别为0.11％、0.06％及0.13％。阿米替林和布比卡因的效价比为0.54(95％CI，0.38-0.71)，阿米替林和利多卡因的效价比为1.18(95％CI，1.52-0.84)。布比卡因和利多卡因的效价比为0.46(95％CI，0.31-0.61)。结论：大鼠IVRA模型可用作药物IVRA效能的测定。阿米替林IVRA效能高于利多卡因，效价比为1.18(95％CI，1.52-0.84)；低于布比卡因，效价比为0.54(95％CI，0.38-0.71)。第三部分阿米替林、布比卡因局部麻醉作用持续时间及局部组织毒性的比较目的：通过和等效浓度的布比卡因比较，探讨阿米替林局部麻醉作用持续时间和局部组织毒性。方法：体重为190—240克的健康成年雄性大鼠32只，随机分为四组，即药物等效浓度实验组：0.22％阿米替林组(A_(2EC50)组，n=8)、0.33％阿米替林组(A_(3EC50)组，n=8)、0.12％布比卡因组(B_(2EC50)组，，n=8)，0.18％布比卡因组(B_(3EC50)组，n=8)。建立大鼠IVRA模型，各组分别给予0.22％阿米替林0.5ml、0.33％阿米替林0.5ml、0.12％布比卡因0.5ml、0.18％布比卡因0.5ml。观察实验当日局部麻醉作用、神经功能恢复时间，并于第二、五、九日进行夹尾反应、TFL测量及鼠尾大体皮肤组织评分。结果： B_(2EC50)组、B_(3EC50)组局部麻醉作用恢复时间分别为38.7±16.8分钟、51.3±17.7分钟；A_(2EC50)组及A_(3EC50)组局部麻醉作用恢复时间分别为232.5±96.4分钟和380.0±127.9分钟，显著长于布比卡因组(P＜0.001)，约为同等效浓度布比卡因的6—7倍。第二、五、九日A_(2EC50)组及B_(2EC50)组、B_(3EC50)组及A_(3EC50)组的5只大鼠：大鼠各段夹尾反应阳性，TFL均在1—4秒内，和基础值比较均无统计学差异(P＞0.05)；鼠尾皮肤完好，皮肤评分为0分。A_(3EC50)组另外3只大鼠夹尾反应和TFL未恢复；皮肤损伤，皮肤评分为6分。结论： 1．阿米替林局部麻醉作用时间长，为同等效浓度布比卡因长的6—7倍。 2．阿米替林组织毒性与浓度相关，0.33％可产生局部神经损伤。小结本研究(1)利用大鼠鼠尾特点，建立新的IVRA模型。应用夹尾反应作为鼠尾局部麻醉作用的判断指标，应用甩尾时间作为鼠尾神经功能恢复的评价指标。大鼠IVRA模型中，通过给予美蓝试剂，证明药物可以通过渗透至止血带以下皮肤组织；通过给予利多卡因，证明局麻药可以产生止血带以下的局部麻醉作用；止血带阻断60分钟未造成鼠尾神经不可逆损伤，阻断30分钟未影响松止血带后的TFL。新建的大鼠IVRA模型有以下优点：①具有临床IVRA的基本特点。②评价指标明确③模型成功率高④简单经济⑤可重复性强等。(2)利用大鼠IVRA模型，采用序贯法得出阿米替林、布比卡因及利多卡因IVRA作用的EC_(50)分别为0.11％、0.06％及0.13％。阿米替林与两种传统的局麻药效价比分别为0.54(95％CI，0.38-0.71)和1.18(95％CI，1.52-0.84)。(3)大鼠IVRA模型中，等效浓度的阿米替林麻醉作用恢复时间显著长于布比卡因(P＜0.001)，约为布比卡因的6-7倍。0.33％阿米替林组3只大鼠出现TFL未恢复，鼠尾皮肤组织损伤，说明①阿米替林和布比卡因相比，虽然IVRA效能较低，但一旦达到有效浓度后，其产生神经阻滞时间明显长于同等效浓度甚至更高浓度的布比卡因。②阿米替林存在浓度相关的局部组织毒性等，0.33％可产生局部组织损伤。综上所述，大鼠IVRA模型简单易行，经济实用，成功率高，可以用作IVRA的药物相关研究等。根据阿米替林作用时间长、效能低、高浓度会产生局部组织毒性的特点，探讨阿米替林和传统的局麻药合用或作为局麻药的辅助药发挥其超长效神经阻滞作用等具有重要意义。
[Abstract]:Background :

Intravenous regional anesthesia
IVRA , also known as Bier Block , began to be used in clinic in 1908 . This method is mainly applicable to adult limb short - term operation , has the advantages of economy , convenience , exact effect , no nerve trauma , etc . It is widely used in animal models such as dogs and rabbits . However , the models are expensive , complex in operation and poor in repeatability . Therefore , it is necessary to establish a new economic and practical animal model .

Amidlin is a kind of traditional tricyclic antidepressants . It is mainly used for treating depression and relieving neuropathic pain . Recently , it has been found that amitripin can produce nerve block similar to traditional Chinese medicine . This study aims to establish the rat IVRA model , and use the model to study the IVRA efficacy , the action time , the local application toxicity , the safety range and so on . The possibility of the application of amitriptimibe in IVRA is discussed , and the theory and experimental basis for clinical application are provided .

The establishment of the IVRA model in the first part of rats

Purpose :

( 1 ) To establish the rat IVRA model ( 2 ) to verify the effect of tourniquet blocking time on the rat tail neurological function in the rat IVRA model .

Method :

( 1 ) Sixteen healthy adult male SD rats were randomly divided into 0.5 % lidocaine group and normal saline control group . The rats were divided into three segments : upper segment , middle segment and last segment .

( 2 ) The rat IVRA model was established in 3 healthy adult male rats ( 1 ) , and 0.5 ml of methylene blue was given . After observation of the color change of the tail skin of rats , the diffusion of the drug in the model was verified .

( 3 ) Twenty - four healthy male rats were randomly divided into four groups : group S _ ( 10 ) , 15 - minute group ( S _ ( 15 ) group ) , 30 - minute group ( S _ ( 30 ) group ) and 60 - minute group ( S _ ( 60 ) group ) .

Results :

( 1 ) In the lidocaine group , only the tail of the rat tail was produced after the administration of the rats , the tail end ( below the tourniquet ) was negative , and the TFL was 10 seconds . The recovery time of the tail of the tail of the tail was 17.5 卤 11.2 min , 26 . 4 卤 19.6 min , respectively .
The recovery time of TFL was 30 . 3 卤 20 . 5 min , 41 . 5 卤 29 . 4 min .

( 2 ) After administration of 3 rats , only the tail skin of the mouse tail below the tourniquet became blue - black .

( 3 ) After four different tourniquet time groups , the tail reaction was positive after the rats were given physiological saline .
There was no statistical difference between the TFL and the basal values in the other groups ( P > 0.05 ) . There was no statistical difference between the TFL and the basal values at the second day of the rats , and the tail skin was intact .

Conclusion :

IVRA in rats is simple , economical , practical and has high success rate . It has the characteristics of IVRA and can be used as a drug - related study of IVRA .

Comparison of the efficacy of the second part of Amidin , bupivacaine and lidocaine IVRA

Purpose :

In this experiment , the effective concentration ( EC _ ( 50 )) of IVRA was studied by sequential method , and compared with traditional local anesthetic bupivacaine and lidocaine , the efficacy of IVRA in amitripis was discussed .

Method :

Ninety - four healthy adult male rats weighing 190 - 240 g were randomly divided into amitriplin group , bupivacaine group and lidocaine group . The model of IVRA was established . The EC _ ( 50 ) value of three drugs was determined by sequential method . The EC _ ( 50 ) value and the drug effect ratio were calculated .

Results :

The potency ratio of amitriplocillin , bupivacaine and lidocaine was 0.11 % , 0.06 % and 0.13 % , respectively . The potency ratio of amitripinide and bupivacaine was 0.54 ( 95 % CI , 0.38 - 0.71 ) , and the potency ratio of amitriplocillin and lidocaine was 1.18 ( 95 % CI , 1.52 - 0.84 ) . The potency ratio of bupivacaine and lidocaine was 0.46 ( 95 % CI , 0.31 - 0.61 ) .

Conclusion :

The IVRA model could be used as the efficacy of IVRA in rats . The efficacy of amitripinidine IVRA was higher than that of lidocaine and the potency ratio was 1.18 ( 95 % CI , 1.52 - 0.84 ) ;
Below bupivacaine , the potency ratio was 0.54 ( 95 % CI , 0.38 - 0.71 ) .

Comparison of the duration of local anesthesia and local tissue toxicity in the third part of amitripin and bupivacaine

Purpose :

The duration of local anesthesia and local tissue toxicity of amitriplocillin were investigated by comparison with bupivacaine in the equivalent concentration .

Method :

Thirty - two healthy adult male rats weighing 190 - 240 g were randomly divided into four groups : group A _ ( 2 _ 50 ) , n = 8 ) , 0 . 33 % ( group A _ ( 3 EC50 ) group , n = 8 ) , 0 . 12 % bupivacaine group ( group B _ ( 2 EC50 ) , n = 8 ) , 0 . 18 % bupivacaine group ( group B _ ( 3 EC50 ) , n = 8 ) . A rat IVRA model was established , and the rats were given 0 . 22 % Amidlin 0.5 ml , 0 . 33 % amoeteine 0.5 ml , 0 . 12 % bupivacaine 0.5 ml , 0 . 18 % bupivacaine 0.5 ml . observe the local anesthetic effect on the day of the experiment , the nerve function recovery time , and the second , fifth and ninth day carry on the tail reaction , the TFL measurement and the rat tail gross skin tissue score .

Results :

The recovery time of local anesthesia in group B _ ( 2EC50 ) group and B _ ( 3EC50 ) group was 38.7 卤 16.8 minutes , 51.3 卤 17.7 min , respectively .
The recovery time of local anesthesia in A _ ( 2EC50 ) group and A _ ( 3EC50 ) group was 232.5 卤 96.4 minutes and 38.0 卤 127.9 minutes , respectively , which was significantly longer than that of bupivacaine group ( P < 0.001 ) , which was about 6 - 7 times of the same equivalent concentration bupivacaine . The second , fifth , nine - day A _ ( 2EC50 ) group and B _ ( 2EC50 ) group , B _ ( 3EC50 ) group , and A _ ( 3EC50 ) group had no statistical difference ( P > 0.05 ) .
The skin was intact and the skin score was 0 . The other 3 rats in the A _ ( 3 EC50 ) group did not recover from the tail reaction and TFL .
Skin injury , skin score of 6 points .

Conclusion :

1 . The local anesthetic duration of amitripin is 6 - 7 times longer than that of bupivacaine .

2 . The toxicity and concentration of amitrip盲lin were correlated , 0.33 % could result in local nerve injury .

small knot

In this study , a new IVRA model was established by using tail - tail characteristics of rats . The tail - tail reaction was used as the index to evaluate the local anesthetic effect of rat tail . The tail - tail time was used as the evaluation index for the recovery of rat tail nerve function .
By administering lidocaine , it is shown that the local anesthetic effect of the local anesthetic can be produced by the local anesthetic .
The results showed that the recovery time of IVRA in rats was significantly longer than that of bupivacaine ( P & lt ; 0.001 ) .
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R614;R-332

【参考文献】