岩静脉离断动物模型的建立及岩静脉离断对兔脑组织的影响

发布时间：2018-01-11 08:24

本文关键词：岩静脉离断动物模型的建立及岩静脉离断对兔脑组织的影响　出处：《中南大学》2010年博士论文　论文类型：学位论文

【摘要】： 背景1929年,Dandy教授在其文章中插图展示了一支走行于桥小脑角区域并经三叉神经背侧汇入岩上窦的静脉,此静脉便是后来众所周知的岩静脉。岩静脉是非常重要且相对恒定的解剖标志,在涉及天幕以及桥小脑角区病变的手术中扮演重要角色。袁贤瑞教授回顾2004年6月至2007年10月进行的听神经瘤手术33例,其中2例手术中岩静脉被电凝处理,31例保护良好。31例岩静脉保护良好的患者术后恢复良好,2例电凝处理岩静脉的患者继发小脑出血性梗死并水肿；1例死亡,1例经后颅窝减压后病情好转。因此,袁贤瑞教授认为手术中应当妥善保护岩静脉,这一观点得到很多学者的支持；但亦有部分学者认为手术中离断岩静脉对术后效果影响不大。目前,文献中未查及关于岩静脉的动物实验研究。目的建立家兔单侧岩静脉离断(双极电凝烧灼后,显微剪剪断)动物模型,观察岩静脉离断后家兔脑组织病理结构、含水量、钠钾离子含量、SOD活力、MDA含量及AQP-4转录与表达。方法1选用健康家兔10只(雌雄不限),解剖分离其颈动脉及颈静脉,应用硅橡胶对新鲜家兔尸头进行灌注。解剖观察灌注好的家兔尸头标本,比较家兔与人类岩静脉的解剖结构。2健康成年家兔144只随机分成两组,即假手术组与岩静脉离断组。每组按处死时间点即手术操作后4h、8h、12h、24h、48h、72h分成六个亚组,每个亚组12只家兔。每个亚组中取6只获取标本后,干湿重法检测小脑及脑干组织含水量；原子火焰吸收分光光度法检测钠钾离子含量；黄嘌呤氧化酶法检测SOD活力；硫代巴比妥酸(TBA)法检测MDA含量,RT-PCR检测AQP-4mRNA转录。亚组中另外六只采用HE染色、尼氏染色及电镜观察脑组织病理结构改变；免疫组织化学检测AQP-4蛋白表达。结果 1家兔尸头灌注血管色泽鲜艳,细小血管显示良好,解剖结构保留完整,其岩静脉解剖结构与人类相似。 2含水量及钠钾离子含量：A小脑组织：假手术组各亚组脑组织含水量、钠钾离子含量无统计学差异(p0.05)；岩静脉离断后4h、8h、12h、24h组脑组织含水量、钠离子含量高于假手术组(p＜0.05),岩静脉离断后4h、8h、12h、24h组脑组织钾离子含量低于假手术组(p0.05)；岩静脉离断后4h脑组织含水量、钠离子含量增加,8-12达高峰,持续至24小时(p＜0.05),钾离子含量自4h降低,8-12小时降至最低(p0.05)。B脑干组织：假手术组各亚组脑组织含水量、钠钾离子含量无统计学差异(p＞0.05)；岩静脉离断后4h脑组织含水量、钠离子含量高于假手术组(p＜0.05),钾离子含量低于假手术组(p＜0.05)。 3小脑组织病理结构改变：假手术组小脑组织神经细胞结构基本正常,岩静脉离断后4h、8h、12h出现水肿,表现为细胞肿胀,尼氏体消失,神经细胞变性坏死等。脑干组织病理结构改变：岩静脉离断4h组出现细胞水肿,岩静脉离断组其余各亚组与假手术组病理结构基本正常。 4 SOD活力检测：A小脑组织：假手术组之间SOD活力无统计学差异(p＞0.05)岩静脉离断后4h、8h、12h、24h脑组织SOD活力低于假手术组(p0.05)岩静脉离断后4h脑组织SOD活力下降,8-12至最低,持续至24小时(p0.05)。B脑干组织：假手术组各亚组脑组织SOD活力无统计学差异(p0.05)岩静脉离断后4h脑组织SOD活力低于假手术组(p0.05)。 5 MDA含量检测：A小脑组织：假手术组之间MDA含量无统计学差异(p0.05)岩静脉离断后4h、8h、12h、24h脑组织MDA含量高于假手术组(p0.05)岩静脉离断后4h脑组织MDA含量上升,8-12达高峰,48-72小时接近假手术组(p0.05)。B脑干组织：假手术组各亚组脑组织MDA含量无统计学差异(p0.05)岩静脉离断后4h脑组织MDA含量高于假手术组(p0.05)。 6 AQP-4转录与表达：A小脑组织：假手术组各亚组脑组织AQP-4转录及表达无统计学差异(p0.05)；岩静脉离断后4h、8h、12h、24h脑组织AQP-4转录与表达增强(p0.05)。B脑干组织：假手术组各亚组脑组织AQP-4转录及表达无统计学差异(p0.05)：岩静脉离断后4h脑组织AQP-4转录及表达强于假手术组(p0.05)。结论 1家兔具有与人类相似的岩静脉结构及引流,从解剖学角度讲,家兔可用于建立岩静脉离断动物模型。 2岩静脉离断组4h、8h、12h、24h亚组小脑组织含水量及钠离子含量较假手术对照组增加,钾离子含量降低。表明岩静脉离断引起钠钾ATP酶功能障碍,导致小脑组织水肿。 3岩静脉离断导致家兔脑组织病理结构改变,此结构改变为可逆性。 4岩静脉离断组4h、8h、12h、24h亚组小脑组织较假手术对照组小脑组织MDA含量增加,SOD活力下降。表明脑细胞代谢及功能障碍,清除氧自由基的能力下降。提示对抗氧自由基可能有助于脑组织功能恢复。 5岩静脉离断组4h、8h、12h、24h亚组小脑组织AQP-4转录与表达较假手术组增强,说明AQP-4参与了脑水肿的形成与发展。提示抑制AQP-4的表达可能能够减轻岩静脉离断导致的脑水肿。
[Abstract]:Background in 1929, Professor Dandy in the illustration showed a walk in the cerebellopontine angle region and the dorsal trigeminal nerve into the superior petrosal sinus vein, the vein is later rock vein. As everyone knows the petrosal vein is a very important anatomic landmark and relatively constant, to play an important role in the bridge and the sky cerebellopontine angle lesions in the operation.
Professor Yuan Xianrui from June 2004 to October 2007 to 33 cases of acoustic neuroma surgery, including 2 cases of surgery in the petrosal vein were electric coagulation treatment, 31 cases of.31 were well protected petrosal vein protection good patients recovered well, 2 cases of edema and infarction electric coagulation of petrosal vein in patients with cerebellar hemorrhage; 1 cases died. 1 cases of posterior fossa decompression after improvement. Therefore, Professor Yuan Xianrui thinks that the petrosal vein should be protected during surgery, this view has been supported by many scholars; but some scholars believe that surgical transection of petrosal vein has little effect on the postoperative effect. At present, the animal experiment study did not check the literature and on the rocks vein.
Objective to establish the rabbit isolated unilateral petrosal vein (bipolar electrocoagulation, micro scissors cut) animal model, observe the petrosal vein from the pathological structure of brain tissue in rabbits after moisture content, sodium and potassium ion and SOD activity, MDA content and expression of AQP-4 transcription.
Methods 1 healthy 10 rabbits (male or female), dissecting the carotid artery and jugular vein, application of silicone rubber are perfused with fresh rabbit cadaver heads. Anatomical observation of rabbit cadaveric head specimens perfused, 144 rats were randomly divided into two groups of.2 healthy adult rabbits were compared with anatomical structure of rabbit and human petrosal vein, i.e. the sham operation group and petrosal vein mutilation group. Each group according to the time of sacrifice is the operation of 4h, 8h, 12h, 24h, 48h, 72h are divided into six sub groups, each sub group of 12 rabbits. Each sub group of 6 specimens were obtained after dry wet weight measurement method in cerebellum and brain tissue water content flame; atomic absorption spectrophotometry to determine the content of sodium and potassium ion; SOD activity was measured with xanthine oxidase method; thiobarbituric acid (TBA) method to detect the content of MDA, RT-PCR detection of AQP-4mRNA subgroup transcription. Another six rabbits with HE staining, Nissl staining and electron microscopy to observe the brain tissue pathological structure Change; immunohistochemical detection of AQP-4 protein expression.
Result
1 the blood vessels in the head of the rabbit were colourful, fine blood vessels were displayed well, the anatomical structure was intact, and the anatomical structure of the veins of the rabbit was similar to that of the human.
2 water and sodium and potassium ion content: A cerebellum: sham operation group, the brain water content, no significant difference in the content of sodium and potassium (P0.05); petrosal vein mutilation of 4h, 8h, 12h, 24h group of brain water content, sodium ion content is higher than that of sham operation group (P < 0.05) petrosal vein mutilation, 4h, 8h, 12h, 24h content of potassium ion in brain tissue were lower than the sham operation group (P0.05); after the mutilation of petrosal vein, 4H content, increase the content of sodium ion, 8-12 reached the peak at 24 hours (P < 0.05), potassium content decreased 4h, 8-12 hours to a minimum (P0.05).B brain stem: sham operation group, the brain water content, no significant difference in the content of sodium and potassium (P > 0.05); after the mutilation of petrosal vein, 4H water, sodium ion content is higher than that of sham operation group (P < 0.05), the potassium content was lower than that of sham operation group (P < 0.05).
3 changes of cerebellar tissue pathological structure: structure of nerve cells of cerebellum were normal in sham operation group, petrosal vein mutilation of 4h, 8h, 12h showed edema, cell swelling, disappearance of Nissl bodies, cell degeneration and necrosis. Pathological changes in the brain stem structure: petrosal vein mutilation of 4H group cells appeared edema, petrosal vein the rest of the break group in each sub group and the pathological structure of sham group was normal.
4 SOD activity detection: A cerebellar tissue: there was no difference between the sham operation group the activity of SOD (P > 0.05) of petrosal vein mutilation of 4h, 8h, 12h, SOD activity in brain tissue of 24h lower than the sham operation group (P0.05) of petrosal vein from SOD activity of brain tissue after 4h 8-12 decreased to the lowest, 24 hours (P0.05).B brain stem: sham operation group SOD activity in each sub group of brain tissue had no significant difference (P0.05) of petrosal vein from the SOD activity of brain tissue after 4H was lower than that of sham operation group (P0.05).
Detection of 5 MDA content: A cerebellar tissues: there was no difference between the sham operation group the content of MDA (P0.05) of petrosal vein mutilation of 4h, 8h, 12h, MDA, 24h content of brain tissue was higher than that of sham group (P0.05) from the petrosal vein MDA in brain tissue after 4H increased, peaked at 8-12, 48-72 hours near sham operation group.B (P0.05) in brain stem: sham operation group MDA in each sub group of brain tissue had no significant difference (P0.05) from the petrosal vein MDA in brain tissue after 4H was higher than that of sham operation group (P0.05).
6: A AQP-4 transcription and expression in cerebellum: sham operation group AQP-4 transcription sub groups of brain tissue and the expression of no significant difference (P0.05); petrosal vein mutilation of 4h, 8h, 12h, enhanced AQP-4 transcription and expression of 24h in brain tissue of brain stem.B (P0.05): sham operation group AQP-4 transcription sub groups of brain tissue and there were no significant difference (P0.05): petrosal vein from AQP-4 brain tissue after 4H transcription and expression in the sham operation group (P0.05).
conclusion
1 the rabbit has the structure and drainage of the vein, which is similar to that of the human. From the anatomic point of view, the rabbit can be used to establish the animal model of the rock vein disconnection.
2, in the 4h, 8h, 12h and 24h subgroups, the content of water and sodium ion in cerebellar tissue was increased compared with the sham operated control group, and the content of potassium decreased.
The changes of the pathological structure of the rabbit brain tissue were caused by 3 venous disconnection, and the structure changed to reversibility.
4, in 4h, 8h, 12h and 24h subgroups of cerebellar vein detachment group, the content of MDA increased and SOD activity decreased in cerebellum tissue compared with sham operated control group. It indicated that the metabolism and dysfunction of brain cells and the ability of scavenging oxygen free radicals decreased. It suggests that anti oxygen free radicals may contribute to the recovery of brain tissue function.
5, the expression and expression of AQP-4 in cerebellar tissue of 4h, 8h, 12h and 24h subgroups were enhanced compared with sham operation group, indicating that AQP-4 was involved in the formation and development of cerebral edema. It suggests that inhibition of AQP-4 expression may reduce cerebral edema caused by disconnection of the petrous vein.

【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R-332

【引证文献】