本文選題：蛛網(wǎng)膜下腔出血　切入點(diǎn)：早期腦損傷　出處：《貴州醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:探討利多卡因(Lidocaine,LD)對(duì)兔蛛網(wǎng)膜下腔出血(subarachnoid hemorrhage,SAH)后早期腦損傷(early brain injury,EBI)的保護(hù)機(jī)制。方法:18只新西蘭大白兔隨機(jī)分為3組:假手術(shù)組(Sham);蛛網(wǎng)膜下腔出血組(SAH);利多卡因組(LD)。采用枕大池一次注血法復(fù)制兔蛛網(wǎng)膜下腔出血模型,各組動(dòng)物稱重后先予咪達(dá)唑侖肌肉注射做基礎(chǔ)麻醉,然后進(jìn)行耳緣靜脈穿刺置管,經(jīng)耳緣靜脈緩慢注入咪達(dá)唑侖、枸櫞酸舒芬太尼注射液及阿托品進(jìn)行全麻誘導(dǎo),待兔全麻生效后立即進(jìn)行氣管插管,氣管插管成功后即可進(jìn)行枕大池穿刺。觸及枕骨下間隙,有突破感后見(jiàn)清亮腦脊液流出,然后采取兔耳中動(dòng)脈血,蛛網(wǎng)膜下腔出血組及利多卡因組動(dòng)物將兔自體動(dòng)脈血1 mL/Kg經(jīng)枕大池注入,假手術(shù)組則注入1 mL/Kg生理鹽水,兔枕大池注射完畢后取30°頭低位30 min,30 min后利多卡因組動(dòng)物再次經(jīng)枕大池注入2%利多卡因0.3 m L,蛛網(wǎng)膜下腔出血組、假手術(shù)組注入0.3 mL生理鹽水,枕大池注液過(guò)程中給予呼吸機(jī)輔助呼吸,全部操作完畢后待兔自然蘇醒,然后送回動(dòng)物實(shí)驗(yàn)中心給予飼養(yǎng)觀察。72 h后對(duì)兔進(jìn)行攝食量和神經(jīng)功能損害分級(jí);采用實(shí)時(shí)熒光定量PCR(RT-PCR)方法及蛋白質(zhì)免疫印跡(Western Blot)方法分別檢測(cè)兔腦海馬組織中含半胱氨酸的天冬氨酸蛋白水解酶3(cysteinyl aspartate specific proteinase 3,Caspase-3)、細(xì)胞色素C(Cytochrome C,Cyt-C)mRNA及蛋白表達(dá)變化;采用免疫組織化學(xué)染色,鏡下觀察兔腦海馬CA1區(qū)Caspase-3、Cyt-C陽(yáng)性表達(dá)情況。結(jié)果:對(duì)兔攝食量和神經(jīng)功能損害分級(jí)發(fā)現(xiàn),與Sham組比較,SAH、LD組動(dòng)物攝食量減少,出現(xiàn)不同程度的神經(jīng)功能損害(P0.05),與SAH組比較,LD組動(dòng)物攝食量及神經(jīng)功能損害無(wú)統(tǒng)計(jì)學(xué)差異(P0.05);RT-PCR、Western Blot統(tǒng)計(jì)結(jié)果顯示,與Sham組比較,SAH、LD組Caspase-3、Cyt-C m RNA及蛋白表達(dá)增加(P0.05),與SAH組比較,LD組Caspase-3、Cyt-C m RNA及蛋白表達(dá)減少(P0.05);免疫組化檢測(cè)Caspase-3、Cyt-C陽(yáng)性細(xì)胞結(jié)果顯示,與Sham組比較,SAH、LD組腦海馬CA1區(qū)Caspase-3、Cyt-C陽(yáng)性細(xì)胞數(shù)增加(P0.05),與SAH組比較,LD組Caspase-3、Cyt-C陽(yáng)性細(xì)胞數(shù)減少(P0.05)。結(jié)論:利多卡因?qū)ν弥刖W(wǎng)膜下腔出血后早期腦損傷的保護(hù)機(jī)制可能與抑制線粒體通路激活有關(guān)。
[Abstract]:Objective: to investigate the protective mechanism of Lidocainedia lidocaine (LDD) on early brain injuryafter subarachnoid hemorrhage (SAH) in rabbits. Methods: eighteen New Zealand white rabbits were randomly divided into 3 groups: sham-operated group, subarachnoid hemorrhage group, Lidocaine group and lidocarcoma group. The rabbit model of subarachnoid hemorrhage was established by single blood injection into the cistern of occipital cistern. After weighing each group, midazolam was injected intramuscularly for basic anesthesia, then the ear margin vein was inserted into the tube, midazolam was injected slowly through the auricular vein, sufentanil citrate injection and atropine were induced by general anesthesia. Tracheal intubation was performed immediately after the rabbit general anesthesia took effect. After the intubation was successful, the cistern cistern was punctured. The suboccipital space was touched, the clear cerebrospinal fluid flowed out after a sense of breakthrough, and then the rabbit middle ear arterial blood was taken. Rabbits in subarachnoid hemorrhage group and lidocaine group were injected with 1 mL/Kg of autologous arterial blood through cistern magnum, while those in sham operation group were injected with 1 mL/Kg normal saline. After 30 擄cistern injection, the lidocaine group was injected with 2% lidocaine (0.3 mL) again through the cistern of the occipital cistern, subarachnoid hemorrhage group and sham operation group (0.3 mL normal saline). During the injection of fluid into the cistern of the occipital cistern, breathing machine was given to the rabbits. After all the operation was completed, the rabbits recovered naturally, and then sent back to the animal experimental center for feeding observation. 72 hours later, the feeding amount and the neurological damage were graded. The expression of 3(cysteinyl aspartate specific proteinase 3 caspase-3, cytochrome C(Cytochrome cyt mRNA and protein in rabbit brain tissues were detected by real-time fluorescence quantitative PCR assay and Western blot assay. Immunohistochemical staining was used to observe the positive expression of Caspase-3 Cyt-C in the CA1 region of rabbit hippocampus under microscope. Results: compared with the Sham group, the consumption of Caspase-3 Cyt-C was decreased in the Sham group. Compared with SAH group, there was no significant difference in food intake and nerve function damage between LD group and SAH group. The results of Western Blot analysis showed that there was no significant difference between LD group and LD group. Compared with Sham group, the expression of Caspase-3 Cyt-C m RNA and protein increased in SAH group, and the expression of Caspase-3 Cyt-C m RNA and protein decreased in LD group compared with SAH group. Compared with the Sham group, the number of Caspase-3 Cyt-C positive cells in the CA1 area of hippocampus increased in the Sham group, and the number of Caspase-3 Cyt-C positive cells decreased in the LD group compared with the SAH group. Conclusion: the protective mechanism of lidocaine on early brain injury after subarachnoid hemorrhage in rabbits may be related to the inhibitory line. Activation of granulocyte pathway is involved.
【學(xué)位授予單位】：貴州醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R743.35

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