血管紧张素Ⅱ系统在家兔神经源性肺水肿中作用的实验研究
发布时间：2018-08-03 07:57
【摘要】：第一部分神经源性肺水肿家兔模型的制备 背景 神经源性肺水肿(Neurogenic pulmonary edema, NPE)是中枢神经系统(Central nervous system, CNS)损伤后急性的威胁生命的并发症,通常在严重CNS损伤后数分钟至数小时内发生。NPE缺乏流行病学数据资料,小儿患者NPE少见,但临床上容易漏诊。近年来,肠道病毒71型(Enterovirus71, EV71)感染所致手足口病成为重要的公众健康问题,重症感染患儿常合并中枢神经系统受累,并发NPE和心肺衰竭者预后不良,病死率、致残率高。 NPE确切的发病机制至今仍未十分明确,主要有冲击伤理论-血流动力性学说和渗透缺陷理论-肺毛细血管通透性学说,目前普遍认为是这两种机制共同作用的结果,交感神经过度兴奋在NPE发生中起重要作用。多年来,国内外研究学者在深入研究其发病机制及治疗的过程中,建立了多种动物模型制备方法。目前,通过EV71感染的方式,可成功复制出中枢神经系统受累模型,但未复制出NPE。因此,本部分研究的目的是选择除EV71感染以外的其他方法,制备神经源性肺水肿模型,同时观察NPE发生过程中交感神经兴奋和血流动力学变化,为后续进行神经源性肺水肿的研究打下基础。 目的 1.复制神经源性肺水肿家兔模型。 2.观察神经源性肺水肿发生过程中交感神经兴奋和血流动力学变化。 方法 1.18只健康成年新西兰兔随机分为3组：正常组6只,NPE组6只,生理盐水组(NS组)6只。 2.NPE模型通过小脑延髓池注射纤维蛋白原和凝血酶诱发形成。 3.家兔麻醉后按分组给予不同处理：①正常组：麻醉后不予其他处理,作为空白对照；②NPE组：麻醉后,进行手术操作(气管插管、PiCCO动静脉导管置管),小脑延髓池穿刺并注射纤维蛋白原和凝血酶；③NS组：麻醉后,进行手术操作(气管插管、PiCCO动静脉导管置管),小脑延髓池穿刺并注射等量生理盐水。 4.实验过程中,正常组观察麻醉后0h、1h、2h、3h、4h、5h、6h呼吸、心率变化；NPE组、NS组,1)观察小脑延髓池穿刺注射药物前后,呼吸(RR)、心率(HR)、平均动脉血压(MABP)的变化及其幅度变化；2)观察不同时点(基础状态B0,小脑延髓池穿刺注射后1min、5min、10min、15min、30min、1h、2h、3h、4h、5h、6h) MABP动态变化；3)分别于基础水平(B0)、注射后15min (P15min)、实验结束时(或P6h)留取血标本,检测血浆肾上腺素(EPI)、去甲肾上腺素(NE)浓度。 5.实验结束后处死动物,立即开胸,观察气管导管有无水肿液自行溢出及肺胸膜下出血程度,评估肺水肿程度,并行组织病理形态学检查。 结果 小脑延髓池注射纤维蛋白原和凝血酶诱导了家兔神经源性肺水肿。各项观察指标变化如下： 1)呼吸指标：各组的基础RR无统计学差异(P0.05)。正常组在观察时间点内,呼吸无明显变化(P0.05)。NPE组、NS组小脑延髓池穿刺注射后即刻(P1min),与基础水平(B0)相比,RR明显增快(P0.01)；两组间RR增快的幅度(△RR)差异无统计学意义。 2)平均动脉血压、心率变化：各组的基础HR无统计学差异(P0.05)。正常组在观察时间点内,心率无明显变化(P0.05)。NPE组、NS组小脑延髓池穿刺注射后即刻(P1min),与B0时比较,MABP升高(P0.01),HR增快(P0.05)；两组间MABP升高的幅度(ΔMABP)差异有统计学意义(P0.01),HR增快的幅度(△HR)差异无统计学意义(P0.05)。NPE组,MABP随时间有下降趋势,短时间内降至基础水平,甚至较低值；NS组,MABP随时间逐渐降至并维持于基础水平。 3)血浆肾上腺素(EPI)、去甲肾上腺素(NE)水平：NPE组和NS组的EPI、 NE基础水平(B0)均无差异(P0.05)。与B0相比,NS组EPI、NE水平在P15min、P6h均无明显变化(P0.05)。与B0相比,NPE组EPI水平在P15min、 P6h均明显升高(P0.01)；NE水平在P15min (P0.01)、P6h升高(P0.05)。NPE组与NS组相比,EPI水平在P15min、P6h均明显升高(P0.01)；NE水平在P15min (P0.01)、P6h升高(P0.05)。 4)气管导管水肿液：实验结束,开胸后不挤压肺,NPE组可见粉红色泡沫状液体自行从气管导管溢出；NS组、正常组均未见气管导管水肿液。 5)肺组织肉眼观：NPE组肺组织明显肿胀,体积增大,边缘变钝,外观暗红,大片淤血,肺切面可见粉红色泡沫状液体。NS组、正常组肺组织无明显肿胀,边缘锐利,外观呈粉红色,无淤血,肺切面未见泡沫状液体。 6)肺胸膜下出血程度：NPE组均有严重的肺水肿,各肺可见Ⅱ-Ⅲ级肺胸膜下出血,其中2/12为Ⅱ级,10/12为Ⅲ级；NS组和正常组均未见肺胸膜下出血,即肺胸膜下出血程度均为0级。 7)肺组织病理形态学：NPE组家兔肺组织肺泡隔明显变宽,部分肺泡可见肺泡隔断裂,肺泡腔融合,红细胞浸润至肺间质或肺泡腔,有的肺泡腔内可见均质淡染的粉红色渗出物。NS组和正常组家兔肺组织肺泡隔正常,肺泡腔完整,未见红细胞浸润,肺泡腔干燥无渗出。 结论 1.小脑延髓池注射纤维蛋白原和凝血酶后,家兔发生交感神经兴奋改变。 2.小脑延髓池注射纤维蛋白原和凝血酶后,家兔出现气管导管水肿液和肺组织病理学肺水肿改变。 3.小脑延髓池注射纤维蛋白原和凝血酶的方法,可用于制备家兔神经源性肺水肿模型。 第二部分AngⅡ系统在家兔神经源性肺水肿中作用的实验研究 背景 近年来研究发现,除了存在一个全身的肾素-血管紧张素系统(Renin-angiotensin system, RAS),机体多个器官和组织也存在独立的局部RAS。研究发现,急性肺损伤时,肺组织局部RAS激活,如急性呼吸窘迫综合征、胎粪吸入综合征、哮喘、严重急性呼吸综合征。因而我们探讨神经源性肺水肿(Neurogenic pulmonary edema, NPE)发生的肺损伤中,局部RAS是否激活,并观察血管紧张素转换酶抑制剂对于NPE的影响,从而为临床防治NPE提供新的突破口。 目的 1.在纤维蛋白诱导的NPE家兔模型中,探讨NPE发生时局部RAS是否激活。 2.给予血管紧张素转换酶抑制剂干预NPE,观察其对NPE的影响。 方法 1.在NPE模型确认的基础上,添加NPE干预组,即依那普利拉组(Ena组),6只健康成年新西兰兔,造模15min后,静脉注射依那普利拉(0.5mg/kg)。实验中纳入的组别为正常组、NPE组、Ena组。 2.实验过程中,观察Ena组不同时点(基础状态B0,小脑延髓池穿刺注射后1min、5min、10min、15min、30min、1h、2h、3h、4h、5h、6h)的呼吸(RR)、平均动脉压(MABP)、心率(HR)的动态变化。 3.实验结束后处死动物,1)立即打开胸腔,观察气管导管有无水肿液自行溢出及肺胸膜下出血情况,评估肺水肿程度；2)留取左肺下叶,常规HE染色行病理组织形态学检查；3)留取右肺组织,测定肺组织Ang Ⅱ浓度及肺组织ACE、ACE2、AT1R mRNA表达水平。 结果 1.肺组织Ang Ⅱ水平NPE组较正常组(539.7±146.6vs253.4±37.2pg/ml)和Ena组(539.7±146.6vs308.7±35.4pg/ml)均增高(P0.05); Ena组与正常组相比,差异无统计学意义(P0.05)。 2.肺组织ACE、ACE2、AT1R mRNA表达水平RT-PCR结果显示： ①ACE mRNA表达水平三组间比较,差异无统计学意义(P0.05),表达水平趋势为NPE组正常组Ena组。 ②ACE2mRNA表达水平与正常组相比,NPE组与Ena组ACE2mRNA表达水平均有下调(P0.05); NPE组与Ena组比较,ACE2mRNA表达水平无差异(P0.05)。 ③AT1R mRNA表达水平三组间比较,差异无统计学意义(P0.05)。 ④ACE/ACE2mRNA表达水平三组间比较,差异无统计学意义(P0.05),趋势为NPE组Ena组正常组。 3.肺水肿评估 1)气管导管水肿液：正常组、Ena组气管导管内均未见粉红色泡沫水肿液自行溢出,NPE组气管导管内可见水肿液。 2)肺胸膜下出血程度：正常组12/12为0级；NPE组2/12为Ⅱ级,10/12为Ⅲ级；Ena组2/12为Ⅰ级,8/12为Ⅱ级,2/12为Ⅲ级。 4.组织病理学肺损伤评分 肺泡过度扩张,Ena组vs NPE组,1.2±0.1vs2.8±0.4分,差异有统计学意义(P0.01)；肺间质水肿,Ena组vs NPE组,1.3±0.2vs2.3±0.5分,差异有统计学意义(P0.01)；肺泡渗出,Ena组vs NPE组,1.5±0.2vs1.8±0.3分,差异无统计学意义(P0.05)。肺损伤总计得分,Ena组vs NPE组,1.3±0.1vs2.3±0.4分,差异有统计学意义(P0.01)。正常组未见肺泡过度扩张、肺间质水肿、肺泡渗出,肺损伤评分为0。 结论 1.NPE发生时,家兔肺组织Ang Ⅱ浓度升高,ACE2mRNA表达下调,肺组织局部RAS激活。 2.依那普利拉干预NPE,家兔肺组织Ang Ⅱ浓度、病理学肺损伤评分降低,提示血管紧张素转换酶抑制剂可能有减轻家兔NPE时肺损伤的作用。
[Abstract]:Part one preparation of rabbit model of neurogenic pulmonary edema
background
Neurogenic pulmonary edema (NPE) is an acute life-threatening complication after the injury of the central nervous system (Central nervous system, CNS)..NPE lacks epidemiological data for several minutes to several hours after severe CNS injury. Children are rarely seen in NPE, but they are easily missed clinically. In recent years, intestines have been found. Hand foot and mouth disease (HMD) caused by Enterovirus71 (Enterovirus71, EV71) infection is an important public health problem. Patients with severe infection often merge with the central nervous system, and the patients with NPE and cardiopulmonary failure have poor prognosis, high mortality and high disability rate.
The exact pathogenesis of NPE is still not very clear, mainly including the theory of impact injury, the theory of hemodynamic and the theory of permeability defect, the theory of pulmonary capillary permeability, which is generally believed to be the result of the joint action of these two mechanisms. The sympathetic excitability plays an important role in the occurrence of NPE. In the process of studying its pathogenesis and treatment, a variety of animal model preparation methods have been established. At present, the central nervous system involvement model can be successfully replicated by EV71 infection, but NPE. has not been replicated. The purpose of this part is to select other methods except EV71 infection to prepare the neurogenic pulmonary edema model. The changes of sympathetic nerve excitation and hemodynamics during the development of NPE were observed to lay a foundation for the follow-up study of neurogenic pulmonary edema.
objective
1. the rabbit model of neurogenic pulmonary edema was replicated.
2. to observe sympathetic excitation and hemodynamic changes in the course of neurogenic pulmonary edema.
Method
1.18 healthy adult New Zealand rabbits were randomly divided into 3 groups: 6 in normal group, 6 in group NPE, and 6 in saline group (group NS).
The 2.NPE model was induced by injection of fibrinogen and thrombin through the cerebellopontine cisterna.
3. rabbits were given different treatments after anesthesia: (1) normal group: after anesthesia, no other treatment, as blank control; group NPE: after anesthesia, operation (tracheal intubation, PiCCO arteriovenous catheterization), cerebellar medullary pool puncture and injection of fibrinogen and thrombin; group NS: after anesthesia, operation (trachea) Intubation, PiCCO arteriovenous catheterization, puncture of cerebellar medullary cistern and injection of equivalent saline.
4. during the experiment, the normal group observed 0h, 1H, 2h, 3h, 4h, 5h, 6h respiration, heart rate change, NPE, NS, 1) before and after the cerebellar medullary cistern puncture injection, respiration (RR), heart rate (HR), mean arterial pressure (MABP) changes and amplitude changes; 2) observation of different points (after cerebellar medullary cistern puncture injection, 10 Min, 15min, 30min, 1H, 2h, 3h, 4h, 5h, 6h) MABP dynamic changes; 3) respectively at the base level (B0), after the injection, at the end of the experiment, to leave the blood samples, to detect the plasma adrenaline, the concentration of norepinephrine.
5. after the end of the experiment, the animals were killed and the chest was opened immediately to observe the self overflow of the tracheal tube and the degree of the hemorrhage of the pulmonary pleura. The degree of pulmonary edema was evaluated and the histopathological examination was performed.
Result
Neurogenic pulmonary edema was induced by injection of fibrinogen and thrombin in cerebellopontine cisterna.
1) respiratory index: there was no statistical difference in the base RR of each group (P0.05). In the normal group, there was no significant change in the respiration (P0.05).NPE group in the observation time point, and the cerebellum medullary pool in NS group was immediately after injection (P1min), and the RR increased significantly (P0.01) compared with the basic level (B0), and there was no significant difference in the amplitude (delta RR) of the RR in the two groups.
2) mean arterial blood pressure and heart rate change: there was no statistical difference in the base HR of each group (P0.05). In the normal group, there was no significant change in heart rate (P0.05).NPE in the observation time point. In group NS, the cerebellar medullary pool was injected immediately after injection (P1min). Compared with B0, MABP increased (P0.01) and HR increased (P0.05); there was a statistical difference between the amplitude of MABP increase (delta) among the two groups. Learning significance (P0.01), the amplitude of HR faster (delta HR) had no statistical significance (P0.05).NPE group, MABP decreased with time, decreased to the base level and even lower in a short time, and MABP in NS group gradually declined and maintained at the base level with time.
3) plasma adrenaline (EPI) and norepinephrine (NE) level: EPI in group NPE and NS group, and NE base level (P0.05). Compared with B0, NS group EPI, NE level is no obvious change. Compared with group NS, EPI levels were significantly higher in P15min and P6h (P0.01); NE levels were higher in P15min (P0.01) and P6h (P0.05).
4) edema fluid of tracheal catheter: the end of the experiment, no squeezing of the lungs after the opening of the chest, and the NPE group can see that the pink foam liquid spilt from the tracheal catheter; in group NS, no tracheal ductus edema fluid is found in the normal group.
5) the naked eye view of lung tissue: the lung tissue of group NPE was obviously swollen, the volume increased, the edge became dull, the appearance was dark red, the blood was red, the pink foam liquid was found in group.NS, the lung tissue of the normal group had no obvious swelling, the edge was sharp, the appearance was pink, no blood, no foam like liquid in the cut face of the lung.
6) the degree of pulmonary and pleural hemorrhage: there were severe pulmonary edema in group NPE, and sub pleural hemorrhage of class II - III in each lung, of which 2/12 was grade II and 10/12 was grade III; no sub pleura hemorrhage was found in group NS and normal group, that is, the degree of sub pleural hemorrhage was grade 0.
7) pathomorphology of lung tissue: pulmonary alveolar septum in NPE group was broadened obviously, alveolar septum was seen in some alveoli, alveolar cavities were fused, red cells infiltrated into pulmonary interstitial or alveolar cavity. In some alveoli, there were homogeneous and light stained pink exudate in.NS group and normal group of pulmonary alveolus in normal group, intact alveolar cavity and no red cells. Infiltration of the alveolar cavity without exudation.
conclusion
1. after injection of fibrinogen and thrombin in the cerebellopontine cisterna, the sympathetic excitation in rabbits changed.
2. After injection of fibrinogen and thrombin into cerebellomedullary cistern, edema of tracheal tube and pulmonary histopathological changes of pulmonary edema occurred in rabbits.
3. the method of injecting fibrinogen and thrombin into cerebellar medullary cistern can be used to prepare rabbit model of neurogenic pulmonary edema.
The second part is the experimental study of the role of Ang II system in rabbit neurogenic pulmonary edema.
background
In recent years, in addition to the existence of a systemic renin angiotensin system (Renin-angiotensin system, RAS), multiple organs and tissues of the body also have independent local RAS. studies. In acute lung injury, the local RAS activation of the lung tissue, such as acute respiratory distress syndrome, meconium aspiration syndrome, asthma, and severe acute respiration, is found. Therefore, we explored the activation of local RAS in the lung injury of Neurogenic pulmonary edema (NPE) and the effect of angiotensin converting enzyme inhibitor on NPE, thus providing a new breakthrough for the clinical prevention and treatment of NPE.
objective
1. in the fibrinogen induced NPE rabbit model, we explored whether local RAS activation occurred in NPE.
2. angiotensin-converting enzyme inhibitor was used to interfere with NPE and observe its effect on NPE.
Method
1. on the basis of NPE model confirmation, the NPE intervention group was added, that is, 6 healthy adult New Zealand rabbits in the enalapril group (group Ena), and after the modeling of 15min, the intravenous injection of enalpli (0.5mg/kg). The group included in the experiment was the normal group, the NPE group, and the Ena group.
2. during the experiment, the dynamic changes of Ena group (basal state B0, 1min, 5min, 10min, 15min, 30min, 1H, 2h, 3h, 4h, 5h, heart rate) were observed.
3. after the end of the experiment, animals were killed, 1) immediately open the thoracic cavity, observe the endotracheal tube with self-propelled fluid overflow and pulmonary pleura hemorrhage, evaluate the degree of pulmonary edema; 2) leave the left lower lobe, routine HE staining, pathological histomorphology examination; 3) leave the right lung tissue, determine the lung tissue Ang II concentration and ACE, ACE2, AT1R mRNA table of lung tissue Up to the level.
Result
1. the level of Ang II in the lung tissue was higher than that of the normal group (539.7 + 146.6vs253.4 + 37.2pg/ml) and Ena group (539.7 + 146.6vs308.7 + 35.4pg/ml) (P0.05), and there was no significant difference between the group Ena and the normal group (P0.05).
2. the expression level of ACE, ACE2 and AT1R mRNA in lung tissue RT-PCR results showed that:
(1) there was no significant difference in the expression level of ACE mRNA between the three groups (P0.05), and the trend of expression was NPE group normal group Ena group.
2. Compared with the normal group, the expression level of ACE2mRNA in group NPE and Ena group was down (P0.05), and there was no difference in the level of ACE2mRNA expression between group NPE and Ena group (P0.05).
(3) there was no significant difference in the expression level of AT1R mRNA between the three groups (P0.05).
(4) there was no significant difference in the level of ACE/ACE2mRNA expression between the three groups (P0.05), and the trend was NPE group Ena group normal group.
3. assessment of pulmonary edema
1) Tracheal catheter edema fluid: Normal group, Ena group tracheal catheter no pink foam edema fluid spontaneous overflow, NPE group tracheal catheter edema fluid can be seen.
2) Degree of pulmonary subpleural hemorrhage: normal group 12/12 was grade 0; NPE group 2/12 was grade II, 10/12 was grade III; Ena group 2/12 was grade I, 8/12 was grade II, 2/12 was grade III.
4. histopathological lung injury score
Pulmonary alveolus overexpansion, group Ena vs NPE group, 1.2 + 0.1vs2.8 + 0.4 points, the difference was statistically significant (P0.01); pulmonary interstitial edema, Ena group vs NPE group, 1.3 + 0.2vs2.3 + 0.5 points, the difference was statistically significant (P0.01); alveolar exudation, Ena group vs groups, 1.5 + 0.3 points, the difference was not statistically significant. Total score of lung injury There was no alveolar hyperdilatation, interstitial edema, alveolar exudation and lung injury score in the normal group.
conclusion
When 1.NPE occurred, the concentration of Ang II in lung tissue increased, the expression of ACE2mRNA was downregulated, and local RAS activation in lung tissue.
2. in NPE, the concentration of Ang II in rabbit lung tissue and the score of pathological lung injury were reduced, suggesting that angiotensin converting enzyme inhibitor may reduce the effect of NPE on lung injury in rabbits.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
[Abstract]:Part one preparation of rabbit model of neurogenic pulmonary edema
background
Neurogenic pulmonary edema (NPE) is an acute life-threatening complication after the injury of the central nervous system (Central nervous system, CNS)..NPE lacks epidemiological data for several minutes to several hours after severe CNS injury. Children are rarely seen in NPE, but they are easily missed clinically. In recent years, intestines have been found. Hand foot and mouth disease (HMD) caused by Enterovirus71 (Enterovirus71, EV71) infection is an important public health problem. Patients with severe infection often merge with the central nervous system, and the patients with NPE and cardiopulmonary failure have poor prognosis, high mortality and high disability rate.
The exact pathogenesis of NPE is still not very clear, mainly including the theory of impact injury, the theory of hemodynamic and the theory of permeability defect, the theory of pulmonary capillary permeability, which is generally believed to be the result of the joint action of these two mechanisms. The sympathetic excitability plays an important role in the occurrence of NPE. In the process of studying its pathogenesis and treatment, a variety of animal model preparation methods have been established. At present, the central nervous system involvement model can be successfully replicated by EV71 infection, but NPE. has not been replicated. The purpose of this part is to select other methods except EV71 infection to prepare the neurogenic pulmonary edema model. The changes of sympathetic nerve excitation and hemodynamics during the development of NPE were observed to lay a foundation for the follow-up study of neurogenic pulmonary edema.
objective
1. the rabbit model of neurogenic pulmonary edema was replicated.
2. to observe sympathetic excitation and hemodynamic changes in the course of neurogenic pulmonary edema.
Method
1.18 healthy adult New Zealand rabbits were randomly divided into 3 groups: 6 in normal group, 6 in group NPE, and 6 in saline group (group NS).
The 2.NPE model was induced by injection of fibrinogen and thrombin through the cerebellopontine cisterna.
3. rabbits were given different treatments after anesthesia: (1) normal group: after anesthesia, no other treatment, as blank control; group NPE: after anesthesia, operation (tracheal intubation, PiCCO arteriovenous catheterization), cerebellar medullary pool puncture and injection of fibrinogen and thrombin; group NS: after anesthesia, operation (trachea) Intubation, PiCCO arteriovenous catheterization, puncture of cerebellar medullary cistern and injection of equivalent saline.
4. during the experiment, the normal group observed 0h, 1H, 2h, 3h, 4h, 5h, 6h respiration, heart rate change, NPE, NS, 1) before and after the cerebellar medullary cistern puncture injection, respiration (RR), heart rate (HR), mean arterial pressure (MABP) changes and amplitude changes; 2) observation of different points (after cerebellar medullary cistern puncture injection, 10 Min, 15min, 30min, 1H, 2h, 3h, 4h, 5h, 6h) MABP dynamic changes; 3) respectively at the base level (B0), after the injection, at the end of the experiment, to leave the blood samples, to detect the plasma adrenaline, the concentration of norepinephrine.
5. after the end of the experiment, the animals were killed and the chest was opened immediately to observe the self overflow of the tracheal tube and the degree of the hemorrhage of the pulmonary pleura. The degree of pulmonary edema was evaluated and the histopathological examination was performed.
Result
Neurogenic pulmonary edema was induced by injection of fibrinogen and thrombin in cerebellopontine cisterna.
1) respiratory index: there was no statistical difference in the base RR of each group (P0.05). In the normal group, there was no significant change in the respiration (P0.05).NPE group in the observation time point, and the cerebellum medullary pool in NS group was immediately after injection (P1min), and the RR increased significantly (P0.01) compared with the basic level (B0), and there was no significant difference in the amplitude (delta RR) of the RR in the two groups.
2) mean arterial blood pressure and heart rate change: there was no statistical difference in the base HR of each group (P0.05). In the normal group, there was no significant change in heart rate (P0.05).NPE in the observation time point. In group NS, the cerebellar medullary pool was injected immediately after injection (P1min). Compared with B0, MABP increased (P0.01) and HR increased (P0.05); there was a statistical difference between the amplitude of MABP increase (delta) among the two groups. Learning significance (P0.01), the amplitude of HR faster (delta HR) had no statistical significance (P0.05).NPE group, MABP decreased with time, decreased to the base level and even lower in a short time, and MABP in NS group gradually declined and maintained at the base level with time.
3) plasma adrenaline (EPI) and norepinephrine (NE) level: EPI in group NPE and NS group, and NE base level (P0.05). Compared with B0, NS group EPI, NE level is no obvious change. Compared with group NS, EPI levels were significantly higher in P15min and P6h (P0.01); NE levels were higher in P15min (P0.01) and P6h (P0.05).
4) edema fluid of tracheal catheter: the end of the experiment, no squeezing of the lungs after the opening of the chest, and the NPE group can see that the pink foam liquid spilt from the tracheal catheter; in group NS, no tracheal ductus edema fluid is found in the normal group.
5) the naked eye view of lung tissue: the lung tissue of group NPE was obviously swollen, the volume increased, the edge became dull, the appearance was dark red, the blood was red, the pink foam liquid was found in group.NS, the lung tissue of the normal group had no obvious swelling, the edge was sharp, the appearance was pink, no blood, no foam like liquid in the cut face of the lung.
6) the degree of pulmonary and pleural hemorrhage: there were severe pulmonary edema in group NPE, and sub pleural hemorrhage of class II - III in each lung, of which 2/12 was grade II and 10/12 was grade III; no sub pleura hemorrhage was found in group NS and normal group, that is, the degree of sub pleural hemorrhage was grade 0.
7) pathomorphology of lung tissue: pulmonary alveolar septum in NPE group was broadened obviously, alveolar septum was seen in some alveoli, alveolar cavities were fused, red cells infiltrated into pulmonary interstitial or alveolar cavity. In some alveoli, there were homogeneous and light stained pink exudate in.NS group and normal group of pulmonary alveolus in normal group, intact alveolar cavity and no red cells. Infiltration of the alveolar cavity without exudation.
conclusion
1. after injection of fibrinogen and thrombin in the cerebellopontine cisterna, the sympathetic excitation in rabbits changed.
2. After injection of fibrinogen and thrombin into cerebellomedullary cistern, edema of tracheal tube and pulmonary histopathological changes of pulmonary edema occurred in rabbits.
3. the method of injecting fibrinogen and thrombin into cerebellar medullary cistern can be used to prepare rabbit model of neurogenic pulmonary edema.
The second part is the experimental study of the role of Ang II system in rabbit neurogenic pulmonary edema.
background
In recent years, in addition to the existence of a systemic renin angiotensin system (Renin-angiotensin system, RAS), multiple organs and tissues of the body also have independent local RAS. studies. In acute lung injury, the local RAS activation of the lung tissue, such as acute respiratory distress syndrome, meconium aspiration syndrome, asthma, and severe acute respiration, is found. Therefore, we explored the activation of local RAS in the lung injury of Neurogenic pulmonary edema (NPE) and the effect of angiotensin converting enzyme inhibitor on NPE, thus providing a new breakthrough for the clinical prevention and treatment of NPE.
objective
1. in the fibrinogen induced NPE rabbit model, we explored whether local RAS activation occurred in NPE.
2. angiotensin-converting enzyme inhibitor was used to interfere with NPE and observe its effect on NPE.
Method
1. on the basis of NPE model confirmation, the NPE intervention group was added, that is, 6 healthy adult New Zealand rabbits in the enalapril group (group Ena), and after the modeling of 15min, the intravenous injection of enalpli (0.5mg/kg). The group included in the experiment was the normal group, the NPE group, and the Ena group.
2. during the experiment, the dynamic changes of Ena group (basal state B0, 1min, 5min, 10min, 15min, 30min, 1H, 2h, 3h, 4h, 5h, heart rate) were observed.
3. after the end of the experiment, animals were killed, 1) immediately open the thoracic cavity, observe the endotracheal tube with self-propelled fluid overflow and pulmonary pleura hemorrhage, evaluate the degree of pulmonary edema; 2) leave the left lower lobe, routine HE staining, pathological histomorphology examination; 3) leave the right lung tissue, determine the lung tissue Ang II concentration and ACE, ACE2, AT1R mRNA table of lung tissue Up to the level.
Result
1. the level of Ang II in the lung tissue was higher than that of the normal group (539.7 + 146.6vs253.4 + 37.2pg/ml) and Ena group (539.7 + 146.6vs308.7 + 35.4pg/ml) (P0.05), and there was no significant difference between the group Ena and the normal group (P0.05).
2. the expression level of ACE, ACE2 and AT1R mRNA in lung tissue RT-PCR results showed that:
(1) there was no significant difference in the expression level of ACE mRNA between the three groups (P0.05), and the trend of expression was NPE group normal group Ena group.
2. Compared with the normal group, the expression level of ACE2mRNA in group NPE and Ena group was down (P0.05), and there was no difference in the level of ACE2mRNA expression between group NPE and Ena group (P0.05).
(3) there was no significant difference in the expression level of AT1R mRNA between the three groups (P0.05).
(4) there was no significant difference in the level of ACE/ACE2mRNA expression between the three groups (P0.05), and the trend was NPE group Ena group normal group.
3. assessment of pulmonary edema
1) Tracheal catheter edema fluid: Normal group, Ena group tracheal catheter no pink foam edema fluid spontaneous overflow, NPE group tracheal catheter edema fluid can be seen.
2) Degree of pulmonary subpleural hemorrhage: normal group 12/12 was grade 0; NPE group 2/12 was grade II, 10/12 was grade III; Ena group 2/12 was grade I, 8/12 was grade II, 2/12 was grade III.
4. histopathological lung injury score
Pulmonary alveolus overexpansion, group Ena vs NPE group, 1.2 + 0.1vs2.8 + 0.4 points, the difference was statistically significant (P0.01); pulmonary interstitial edema, Ena group vs NPE group, 1.3 + 0.2vs2.3 + 0.5 points, the difference was statistically significant (P0.01); alveolar exudation, Ena group vs groups, 1.5 + 0.3 points, the difference was not statistically significant. Total score of lung injury There was no alveolar hyperdilatation, interstitial edema, alveolar exudation and lung injury score in the normal group.
conclusion
When 1.NPE occurred, the concentration of Ang II in lung tissue increased, the expression of ACE2mRNA was downregulated, and local RAS activation in lung tissue.
2. in NPE, the concentration of Ang II in rabbit lung tissue and the score of pathological lung injury were reduced, suggesting that angiotensin converting enzyme inhibitor may reduce the effect of NPE on lung injury in rabbits.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.6
【相似文献】
相关期刊论文 前10条
1 左佑!453100河南卫辉,周文科!453100河南卫辉,周杰!453100河南卫辉,惠磊!453100河南卫辉,冯光!453100河南卫辉;脑外伤并发神经源性肺水肿10例[J];新乡医学院学报;1999年04期
2 胡华成;过中方;诸荣恩;;神经源性肺水肿[J];国外医学.内科学分册;1980年03期
3 章永伟,邹洁静;颅脑损伤并发神经源性肺水肿的护理体会[J];第四军医大学吉林军医学院学报;1995年02期
4 郑少钦,杨应明;神经源性肺水肿的病因、诊断与治疗[J];医师进修杂志;1999年07期
5 尹康,朱良才,腾灵方,金玲江,彭余江,林军挺,宋维康,姜爱国;颅脑损伤继发神经源性肺水肿的临床研究[J];青海医药杂志;1999年07期
6 王红梅,巩新华;颅脑损伤并发神经源性肺水肿的观察与护理[J];齐鲁护理杂志;2000年02期
7 周文科,周国胜,周杰,左佑,冯光,黄立勇,史耀亭,张新中;脑外伤并发神经源性肺水肿的防治[J];中国急救医学;2001年02期
8 张义举,孟良,周亚梅;出血性脑血管疾病并发神经源性肺水肿11例临床观察[J];牡丹江医学院学报;2001年02期
9 黄登煌;急性颅脑损伤并发神经源性肺水肿8例报告[J];交通医学;1997年03期
10 彭燕;急性脑血管病并发神经源性肺水肿40例临床分析[J];河南实用神经疾病杂志;1999年05期
相关会议论文 前10条
1 郭万华;谷子林;刘亚娟;王增利;;野生凬草对家兔生产性能及血清生化指标的的影响[A];京津冀畜牧兽医科技创新交流会暨新思想、新观点、新方法论坛论文集[C];2008年
2 乔西波;栾新;李标;姜运良;樊新忠;;家兔MSTN基因SNPs及其与肉用性能关系的研究[A];中国动物遗传育种研究进展——第十五次全国动物遗传育种学术讨论会论文集[C];2009年
3 周红梅;郭晓令;徐宁迎;;C.elegans fat-1基因克隆及其家兔表达载体构建[A];中国动物遗传育种研究进展——第十五次全国动物遗传育种学术讨论会论文集[C];2009年
4 R塷"}3;王，
本文编号：2161105
本文链接：https://www.wllwen.com/yixuelunwen/eklw/2161105.html
最近更新
教材专著
