乙醇對家兔腦基底動(dòng)脈影響的研究
發(fā)布時(shí)間：2018-07-23 14:26
【摘要】： 研究背景在法醫(yī)實(shí)踐中，經(jīng)常遇到蛛網(wǎng)膜下腔出血而導(dǎo)致死亡的案例，這類案件常發(fā)生在飲酒后的糾紛事件中，這就導(dǎo)致了在法醫(yī)學(xué)實(shí)際鑒定中，外力、酒精含量及疾病對蛛網(wǎng)膜下腔出血的作用往往成為一個(gè)爭論的焦點(diǎn)，其原因是我們不清楚其間的相互關(guān)系和確切的作用機(jī)制，因此提供不出一個(gè)科學(xué)的診斷標(biāo)準(zhǔn)。近年來，生物力學(xué)的研究方法和手段，為我們解決這方面的問題提供了一種新的思路和方法。 目的本研究以不同的酒精濃度，對家兔灌酒，測定血中酒精濃度，建立血液乙醇濃度梯度動(dòng)物模型，測定不同乙醇濃度動(dòng)物模型局部腦微區(qū)的血流灌注量改變。在此基礎(chǔ)上測定不同酒精濃度下家兔腦基底動(dòng)脈的零壓力狀態(tài)，并用免疫組化的方法檢測血管內(nèi)皮細(xì)胞HSP70的表達(dá)，以探討乙醇對腦基底動(dòng)脈的影響，從而探索運(yùn)用生物力學(xué)的方法為法醫(yī)學(xué)實(shí)踐中解決諸如醉酒在蛛網(wǎng)膜下腔出血中的作用等案件提供一定的理論基礎(chǔ)。 材料與方法 1．動(dòng)物分組及血液乙醇濃度梯度家兔動(dòng)物模型的建立家兔40只，隨機(jī)分成4組。用酒精灌胃造模，按0ml／Kg、5ml／Kg、10ml／Kg、15ml／Kg給家兔灌注56％市售食用白酒。 2．家兔灌胃后不同時(shí)間血液酒精濃度的測定 采用上海天美公司生產(chǎn)的D-7900氣相色譜儀-美國DANNI86.85頂空自動(dòng)進(jìn)樣器檢測各組家兔灌胃后30 min、60 min、75 min、90min、105min、120min血液酒精濃度。 3．家兔灌胃后不同時(shí)間測定腦血流量 采用瑞典Perimed公司生產(chǎn)的PeriFlux5000激光多譜勒血流儀檢測各組家兔灌胃前及灌胃后5 min、15 min、30 min、60 min腦微區(qū)血流量情況。 4．基底動(dòng)脈內(nèi)皮細(xì)胞熱休克蛋白(HSP)70的表達(dá)情況 取灌胃2小時(shí)后家兔基底動(dòng)脈，做組織切片，用免疫組織化學(xué)的方法染色，檢測熱休克蛋白(HSP)70的表達(dá)。 5．家兔基底動(dòng)脈零應(yīng)力狀態(tài)的測定 各組家兔酒精灌胃2小時(shí)后，開顱取出家兔基底動(dòng)脈，沿血管長軸連續(xù)橫切，得到長度約為1mm的血管環(huán)，然后沿血管環(huán)徑向剪開血管，靜置于kreb's液中20分鐘，用數(shù)碼相機(jī)采集得到血管零應(yīng)力狀態(tài)的數(shù)字化幾何信息。用計(jì)算機(jī)圖像分析軟件AutoCAD Electrical 2006測量基底動(dòng)脈張開角大小，以了解血管零應(yīng)力狀態(tài)的變化。 結(jié)果 1．灌酒后血液乙醇濃度持續(xù)增高，灌胃后1.5-2h，血液乙醇濃度達(dá)到峰值；灌酒后血液乙醇濃度與灌酒量存在正相關(guān)關(guān)系； 2．家兔灌酒后局部腦微區(qū)血流量值明顯增大，至30分鐘后，各組家兔腦血流量基本恒定；灌酒后局部腦微區(qū)血流量值與灌酒量存在正相關(guān)關(guān)系； 3．灌酒2小時(shí)后，家兔基底動(dòng)脈切片，做免疫組織化學(xué)染色，，發(fā)現(xiàn)熱休克蛋白(HSP)70的表達(dá)，隨灌酒量的增加而增加(P＜0.05)； 4．各組家兔酒精灌胃2小時(shí)后基底動(dòng)脈張開角均明顯降低，各組之間差異存在統(tǒng)計(jì)學(xué)意義(P＜0.05)，且隨灌胃量的增大基底動(dòng)脈張開角降低越大。說明酒精濃度越大，基底動(dòng)脈零應(yīng)力狀態(tài)越低。 結(jié)論本研究通過實(shí)驗(yàn)證明：①灌酒后血液乙醇濃度與灌酒量存在正相關(guān)關(guān)系；②灌酒后局部腦微區(qū)血流量值與灌酒量存在正相關(guān)關(guān)系；③HSP70的表達(dá)隨灌酒量的增加而增加；④酒精濃度越高，腦基底動(dòng)脈零應(yīng)力狀態(tài)越低。
[Abstract]:Background in forensic practice, cases of death caused by subarachnoid hemorrhage are often encountered. These cases often occur in post alcohol disputes. This leads to the effect of external force, alcohol content and disease on subarachnoid hemorrhage in forensic forensic identification, which is often the focus of argument. The reason is that we In recent years, the research methods and means of biomechanics have provided us with a new way of thinking and methods for solving the problems in this field.
Objective to study the blood concentration of alcohol in the blood and establish a blood alcohol concentration gradient animal model with different alcohol concentration, and to determine the change of blood flow perfusion in the local brain microregion of the animal models with different ethanol concentration. The expression of HSP70 in vascular endothelial cells was detected in order to explore the effect of ethanol on the cerebral basilar artery, and to explore the application of biomechanics to provide a theoretical basis for solving cases such as the effect of drunken alcohol in the subarachnoid hemorrhage in forensic practice.
Materials and methods
1. the rabbits were divided into 4 groups, and 40 rabbits were randomly divided into 4 groups. The rabbits were randomly divided into 4 groups. The rabbits were perfused by alcohol to make the model, and 56% municipal liquor was perfused to the rabbits by 0ml / Kg, 5ml / Kg, 10ml / Kg and 15ml / Kg.
2. determination of blood alcohol concentration in rabbits after different times of gavage
The blood alcohol concentration of 30 min, 60 min, 75 min, 90min, 105min, 120min in each group of rabbits was detected by D-7900 gas chromatograph produced by Shanghai Tian Mei company and American DANNI86.85 headspace automatic injector.
The determination of cerebral blood flow in 3. rabbits at different time after gavage
The blood flow of 5 min, 15 min, 30 min, and 60 min brain microregion before and after intragastric perfusion was detected by PeriFlux5000 laser Doppler flow meter produced by Swedish Perimed company.
4. expression of heat shock protein (HSP) 70 in basilar artery endothelial cells
After 2 hours of gavage, the basilar artery of rabbits was sectioning, and the expression of heat shock protein (HSP) 70 was detected by immunohistochemical staining.
Determination of the zero stress state of basilar artery in 5. rabbits
After 2 hours of alcohol irrigated by the rabbits, the basilar artery was removed by craniotomy, and the vascular ring was cut along the long axis of the blood vessel. The vascular ring length about 1mm was obtained, then the blood vessels were cut open along the ring of blood vessel, and the digital geometric information of the zero stress state of the blood vessel was collected by the digital camera for 20 minutes. The computer image analysis software A was used. UtoCAD Electrical 2006 measured the opening angle of the basilar artery to understand the change of the vessel's zero stress state.
Result
1. after irrigation, the concentration of alcohol in blood continued to increase, 1.5-2h after gavage, the concentration of blood ethanol reached its peak value, and there was a positive correlation between the concentration of alcohol and the amount of alcohol after irrigation.
2. after drinking alcohol, the blood flow rate of local cerebral microregion increased obviously. After 30 minutes, the cerebral blood flow of the rabbits in each group was basically constant, and there was a positive correlation between the blood flow value of local brain microarea and the amount of wine after drinking.
3. after 2 hours of drinking, the expression of heat shock protein (HSP) 70 increased with the increase of the amount of drinking alcohol (P < 0.05).
4. the basilar artery opening angle decreased obviously after 2 hours of alcohol gavage, and the difference between each group was statistically significant (P < 0.05), and the larger the basilar artery opening angle decreased with the increase of gastric perfusion, the higher the alcohol concentration, the lower the zero stress state of the basilar artery.
Conclusion the results of the study show that: (1) there is a positive correlation between the concentration of alcohol and the amount of alcohol after drinking; (2) there is a positive correlation between the blood flow value of the local brain microarea and the amount of drinking wine after drinking alcohol; (3) the expression of HSP70 increases with the increase of the amount of alcohol; the higher the alcohol concentration, the lower the zero stress state of the cerebral basilar artery.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：D919
[Abstract]:Background in forensic practice, cases of death caused by subarachnoid hemorrhage are often encountered. These cases often occur in post alcohol disputes. This leads to the effect of external force, alcohol content and disease on subarachnoid hemorrhage in forensic forensic identification, which is often the focus of argument. The reason is that we In recent years, the research methods and means of biomechanics have provided us with a new way of thinking and methods for solving the problems in this field.
Objective to study the blood concentration of alcohol in the blood and establish a blood alcohol concentration gradient animal model with different alcohol concentration, and to determine the change of blood flow perfusion in the local brain microregion of the animal models with different ethanol concentration. The expression of HSP70 in vascular endothelial cells was detected in order to explore the effect of ethanol on the cerebral basilar artery, and to explore the application of biomechanics to provide a theoretical basis for solving cases such as the effect of drunken alcohol in the subarachnoid hemorrhage in forensic practice.
Materials and methods
1. the rabbits were divided into 4 groups, and 40 rabbits were randomly divided into 4 groups. The rabbits were randomly divided into 4 groups. The rabbits were perfused by alcohol to make the model, and 56% municipal liquor was perfused to the rabbits by 0ml / Kg, 5ml / Kg, 10ml / Kg and 15ml / Kg.
2. determination of blood alcohol concentration in rabbits after different times of gavage
The blood alcohol concentration of 30 min, 60 min, 75 min, 90min, 105min, 120min in each group of rabbits was detected by D-7900 gas chromatograph produced by Shanghai Tian Mei company and American DANNI86.85 headspace automatic injector.
The determination of cerebral blood flow in 3. rabbits at different time after gavage
The blood flow of 5 min, 15 min, 30 min, and 60 min brain microregion before and after intragastric perfusion was detected by PeriFlux5000 laser Doppler flow meter produced by Swedish Perimed company.
4. expression of heat shock protein (HSP) 70 in basilar artery endothelial cells
After 2 hours of gavage, the basilar artery of rabbits was sectioning, and the expression of heat shock protein (HSP) 70 was detected by immunohistochemical staining.
Determination of the zero stress state of basilar artery in 5. rabbits
After 2 hours of alcohol irrigated by the rabbits, the basilar artery was removed by craniotomy, and the vascular ring was cut along the long axis of the blood vessel. The vascular ring length about 1mm was obtained, then the blood vessels were cut open along the ring of blood vessel, and the digital geometric information of the zero stress state of the blood vessel was collected by the digital camera for 20 minutes. The computer image analysis software A was used. UtoCAD Electrical 2006 measured the opening angle of the basilar artery to understand the change of the vessel's zero stress state.
Result
1. after irrigation, the concentration of alcohol in blood continued to increase, 1.5-2h after gavage, the concentration of blood ethanol reached its peak value, and there was a positive correlation between the concentration of alcohol and the amount of alcohol after irrigation.
2. after drinking alcohol, the blood flow rate of local cerebral microregion increased obviously. After 30 minutes, the cerebral blood flow of the rabbits in each group was basically constant, and there was a positive correlation between the blood flow value of local brain microarea and the amount of wine after drinking.
3. after 2 hours of drinking, the expression of heat shock protein (HSP) 70 increased with the increase of the amount of drinking alcohol (P < 0.05).
4. the basilar artery opening angle decreased obviously after 2 hours of alcohol gavage, and the difference between each group was statistically significant (P < 0.05), and the larger the basilar artery opening angle decreased with the increase of gastric perfusion, the higher the alcohol concentration, the lower the zero stress state of the basilar artery.
Conclusion the results of the study show that: (1) there is a positive correlation between the concentration of alcohol and the amount of alcohol after drinking; (2) there is a positive correlation between the blood flow value of the local brain microarea and the amount of drinking wine after drinking alcohol; (3) the expression of HSP70 increases with the increase of the amount of alcohol; the higher the alcohol concentration, the lower the zero stress state of the cerebral basilar artery.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：D919
【參考文獻(xiàn)】
相關(guān)期刊論文 前10條
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