巖靜脈離斷動(dòng)物模型的建立及巖靜脈離斷對(duì)兔腦組織的影響
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本文關(guān)鍵詞:巖靜脈離斷動(dòng)物模型的建立及巖靜脈離斷對(duì)兔腦組織的影響 出處:《中南大學(xué)》2010年博士論文 論文類(lèi)型:學(xué)位論文
更多相關(guān)文章: 巖靜脈 含水量 病理結(jié)構(gòu) MDA SOD AQP-4
【摘要】: 背景1929年,Dandy教授在其文章中插圖展示了一支走行于橋小腦角區(qū)域并經(jīng)三叉神經(jīng)背側(cè)匯入巖上竇的靜脈,此靜脈便是后來(lái)眾所周知的巖靜脈。巖靜脈是非常重要且相對(duì)恒定的解剖標(biāo)志,在涉及天幕以及橋小腦角區(qū)病變的手術(shù)中扮演重要角色。 袁賢瑞教授回顧2004年6月至2007年10月進(jìn)行的聽(tīng)神經(jīng)瘤手術(shù)33例,其中2例手術(shù)中巖靜脈被電凝處理,31例保護(hù)良好。31例巖靜脈保護(hù)良好的患者術(shù)后恢復(fù)良好,2例電凝處理巖靜脈的患者繼發(fā)小腦出血性梗死并水腫;1例死亡,1例經(jīng)后顱窩減壓后病情好轉(zhuǎn)。因此,袁賢瑞教授認(rèn)為手術(shù)中應(yīng)當(dāng)妥善保護(hù)巖靜脈,這一觀點(diǎn)得到很多學(xué)者的支持;但亦有部分學(xué)者認(rèn)為手術(shù)中離斷巖靜脈對(duì)術(shù)后效果影響不大。目前,文獻(xiàn)中未查及關(guān)于巖靜脈的動(dòng)物實(shí)驗(yàn)研究。 目的建立家兔單側(cè)巖靜脈離斷(雙極電凝燒灼后,顯微剪剪斷)動(dòng)物模型,觀察巖靜脈離斷后家兔腦組織病理結(jié)構(gòu)、含水量、鈉鉀離子含量、SOD活力、MDA含量及AQP-4轉(zhuǎn)錄與表達(dá)。 方法1選用健康家兔10只(雌雄不限),解剖分離其頸動(dòng)脈及頸靜脈,應(yīng)用硅橡膠對(duì)新鮮家兔尸頭進(jìn)行灌注。解剖觀察灌注好的家兔尸頭標(biāo)本,比較家兔與人類(lèi)巖靜脈的解剖結(jié)構(gòu)。2健康成年家兔144只隨機(jī)分成兩組,即假手術(shù)組與巖靜脈離斷組。每組按處死時(shí)間點(diǎn)即手術(shù)操作后4h、8h、12h、24h、48h、72h分成六個(gè)亞組,每個(gè)亞組12只家兔。每個(gè)亞組中取6只獲取標(biāo)本后,干濕重法檢測(cè)小腦及腦干組織含水量;原子火焰吸收分光光度法檢測(cè)鈉鉀離子含量;黃嘌呤氧化酶法檢測(cè)SOD活力;硫代巴比妥酸(TBA)法檢測(cè)MDA含量,RT-PCR檢測(cè)AQP-4mRNA轉(zhuǎn)錄。亞組中另外六只采用HE染色、尼氏染色及電鏡觀察腦組織病理結(jié)構(gòu)改變;免疫組織化學(xué)檢測(cè)AQP-4蛋白表達(dá)。 結(jié)果 1家兔尸頭灌注血管色澤鮮艷,細(xì)小血管顯示良好,解剖結(jié)構(gòu)保留完整,其巖靜脈解剖結(jié)構(gòu)與人類(lèi)相似。 2含水量及鈉鉀離子含量:A小腦組織:假手術(shù)組各亞組腦組織含水量、鈉鉀離子含量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05);巖靜脈離斷后4h、8h、12h、24h組腦組織含水量、鈉離子含量高于假手術(shù)組(p<0.05),巖靜脈離斷后4h、8h、12h、24h組腦組織鉀離子含量低于假手術(shù)組(p0.05);巖靜脈離斷后4h腦組織含水量、鈉離子含量增加,8-12達(dá)高峰,持續(xù)至24小時(shí)(p<0.05),鉀離子含量自4h降低,8-12小時(shí)降至最低(p0.05)。B腦干組織:假手術(shù)組各亞組腦組織含水量、鈉鉀離子含量無(wú)統(tǒng)計(jì)學(xué)差異(p>0.05);巖靜脈離斷后4h腦組織含水量、鈉離子含量高于假手術(shù)組(p<0.05),鉀離子含量低于假手術(shù)組(p<0.05)。 3小腦組織病理結(jié)構(gòu)改變:假手術(shù)組小腦組織神經(jīng)細(xì)胞結(jié)構(gòu)基本正常,巖靜脈離斷后4h、8h、12h出現(xiàn)水腫,表現(xiàn)為細(xì)胞腫脹,尼氏體消失,神經(jīng)細(xì)胞變性壞死等。腦干組織病理結(jié)構(gòu)改變:巖靜脈離斷4h組出現(xiàn)細(xì)胞水腫,巖靜脈離斷組其余各亞組與假手術(shù)組病理結(jié)構(gòu)基本正常。 4 SOD活力檢測(cè):A小腦組織:假手術(shù)組之間SOD活力無(wú)統(tǒng)計(jì)學(xué)差異(p>0.05)巖靜脈離斷后4h、8h、12h、24h腦組織SOD活力低于假手術(shù)組(p0.05)巖靜脈離斷后4h腦組織SOD活力下降,8-12至最低,持續(xù)至24小時(shí)(p0.05)。B腦干組織:假手術(shù)組各亞組腦組織SOD活力無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)巖靜脈離斷后4h腦組織SOD活力低于假手術(shù)組(p0.05)。 5 MDA含量檢測(cè):A小腦組織:假手術(shù)組之間MDA含量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)巖靜脈離斷后4h、8h、12h、24h腦組織MDA含量高于假手術(shù)組(p0.05)巖靜脈離斷后4h腦組織MDA含量上升,8-12達(dá)高峰,48-72小時(shí)接近假手術(shù)組(p0.05)。B腦干組織:假手術(shù)組各亞組腦組織MDA含量無(wú)統(tǒng)計(jì)學(xué)差異(p0.05)巖靜脈離斷后4h腦組織MDA含量高于假手術(shù)組(p0.05)。 6 AQP-4轉(zhuǎn)錄與表達(dá):A小腦組織:假手術(shù)組各亞組腦組織AQP-4轉(zhuǎn)錄及表達(dá)無(wú)統(tǒng)計(jì)學(xué)差異(p0.05);巖靜脈離斷后4h、8h、12h、24h腦組織AQP-4轉(zhuǎn)錄與表達(dá)增強(qiáng)(p0.05)。B腦干組織:假手術(shù)組各亞組腦組織AQP-4轉(zhuǎn)錄及表達(dá)無(wú)統(tǒng)計(jì)學(xué)差異(p0.05):巖靜脈離斷后4h腦組織AQP-4轉(zhuǎn)錄及表達(dá)強(qiáng)于假手術(shù)組(p0.05)。 結(jié)論 1家兔具有與人類(lèi)相似的巖靜脈結(jié)構(gòu)及引流,從解剖學(xué)角度講,家兔可用于建立巖靜脈離斷動(dòng)物模型。 2巖靜脈離斷組4h、8h、12h、24h亞組小腦組織含水量及鈉離子含量較假手術(shù)對(duì)照組增加,鉀離子含量降低。表明巖靜脈離斷引起鈉鉀ATP酶功能障礙,導(dǎo)致小腦組織水腫。 3巖靜脈離斷導(dǎo)致家兔腦組織病理結(jié)構(gòu)改變,此結(jié)構(gòu)改變?yōu)榭赡嫘浴?4巖靜脈離斷組4h、8h、12h、24h亞組小腦組織較假手術(shù)對(duì)照組小腦組織MDA含量增加,SOD活力下降。表明腦細(xì)胞代謝及功能障礙,清除氧自由基的能力下降。提示對(duì)抗氧自由基可能有助于腦組織功能恢復(fù)。 5巖靜脈離斷組4h、8h、12h、24h亞組小腦組織AQP-4轉(zhuǎn)錄與表達(dá)較假手術(shù)組增強(qiáng),說(shuō)明AQP-4參與了腦水腫的形成與發(fā)展。提示抑制AQP-4的表達(dá)可能能夠減輕巖靜脈離斷導(dǎo)致的腦水腫。
[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.
【學(xué)位授予單位】:中南大學(xué)
【學(xué)位級(jí)別】:博士
【學(xué)位授予年份】:2010
【分類(lèi)號(hào)】:R-332
【引證文獻(xiàn)】
相關(guān)期刊論文 前1條
1 成磊;袁賢瑞;;巖靜脈的臨床意義[J];中華神經(jīng)外科疾病研究雜志;2012年06期
,本文編號(hào):1408828
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