乙醇对家兔脑基底动脉影响的研究
发布时间：2018-07-23 14:26
【摘要】： 研究背景在法医实践中，经常遇到蛛网膜下腔出血而导致死亡的案例，这类案件常发生在饮酒后的纠纷事件中，这就导致了在法医学实际鉴定中，外力、酒精含量及疾病对蛛网膜下腔出血的作用往往成为一个争论的焦点，其原因是我们不清楚其间的相互关系和确切的作用机制，因此提供不出一个科学的诊断标准。近年来，生物力学的研究方法和手段，为我们解决这方面的问题提供了一种新的思路和方法。 目的本研究以不同的酒精浓度，对家兔灌酒，测定血中酒精浓度，建立血液乙醇浓度梯度动物模型，测定不同乙醇浓度动物模型局部脑微区的血流灌注量改变。在此基础上测定不同酒精浓度下家兔脑基底动脉的零压力状态，并用免疫组化的方法检测血管内皮细胞HSP70的表达，以探讨乙醇对脑基底动脉的影响，从而探索运用生物力学的方法为法医学实践中解决诸如醉酒在蛛网膜下腔出血中的作用等案件提供一定的理论基础。 材料与方法 1．动物分组及血液乙醇浓度梯度家兔动物模型的建立家兔40只，随机分成4组。用酒精灌胃造模，按0ml／Kg、5ml／Kg、10ml／Kg、15ml／Kg给家兔灌注56％市售食用白酒。 2．家兔灌胃后不同时间血液酒精浓度的测定 采用上海天美公司生产的D-7900气相色谱仪-美国DANNI86.85顶空自动进样器检测各组家兔灌胃后30 min、60 min、75 min、90min、105min、120min血液酒精浓度。 3．家兔灌胃后不同时间测定脑血流量 采用瑞典Perimed公司生产的PeriFlux5000激光多谱勒血流仪检测各组家兔灌胃前及灌胃后5 min、15 min、30 min、60 min脑微区血流量情况。 4．基底动脉内皮细胞热休克蛋白(HSP)70的表达情况 取灌胃2小时后家兔基底动脉，做组织切片，用免疫组织化学的方法染色，检测热休克蛋白(HSP)70的表达。 5．家兔基底动脉零应力状态的测定 各组家兔酒精灌胃2小时后，开颅取出家兔基底动脉，沿血管长轴连续横切，得到长度约为1mm的血管环，然后沿血管环径向剪开血管，静置于kreb's液中20分钟，用数码相机采集得到血管零应力状态的数字化几何信息。用计算机图像分析软件AutoCAD Electrical 2006测量基底动脉张开角大小，以了解血管零应力状态的变化。 结果 1．灌酒后血液乙醇浓度持续增高，灌胃后1.5-2h，血液乙醇浓度达到峰值；灌酒后血液乙醇浓度与灌酒量存在正相关关系； 2．家兔灌酒后局部脑微区血流量值明显增大，至30分钟后，各组家兔脑血流量基本恒定；灌酒后局部脑微区血流量值与灌酒量存在正相关关系； 3．灌酒2小时后，家兔基底动脉切片，做免疫组织化学染色，，发现热休克蛋白(HSP)70的表达，随灌酒量的增加而增加(P＜0.05)； 4．各组家兔酒精灌胃2小时后基底动脉张开角均明显降低，各组之间差异存在统计学意义(P＜0.05)，且随灌胃量的增大基底动脉张开角降低越大。说明酒精浓度越大，基底动脉零应力状态越低。 结论本研究通过实验证明：①灌酒后血液乙醇浓度与灌酒量存在正相关关系；②灌酒后局部脑微区血流量值与灌酒量存在正相关关系；③HSP70的表达随灌酒量的增加而增加；④酒精浓度越高，脑基底动脉零应力状态越低。
[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.
【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】：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.
【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】：D919
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