活体荧光显微镜技术在观察肝硬化门静脉高压大鼠肝脏微循环结构变化中的应用  

Application of intravital fluorescence microscopy in the observation of the changes of hepatic microcircula- tion in rat with hepatic cirrhosis and portal hypertension

作　　者：胡良硕[1] 王浩华[1] 王建华[2] 吕毅[1] 

机构地区：[1]西安交通大学医学院第一附属医院肝胆外科,710061 [2]澳大利亚2145,悉尼大学维斯敏医院胃肠和肝脏内科

出　　处：《中华消化外科杂志》2014年第4期286-290,共5页Chinese Journal of Digestive Surgery

基　　金：卫生部部属（管）医院临床学科重点项目（105）

摘　　要：目的探讨活体荧光显微镜技术观察肝硬化门静脉高压模型大鼠的肝脏微循环结构变化的应用效果。方法取雄性SD大鼠70只，将其中40只SD大鼠采用随机数字表法分为假手术组、胆总管结扎（BDL）模型2周组、BDL模型4周组及BDL模型6周组，每组10只，应用活体荧光显微镜技术观察大鼠肝脏微循环结构变化。将余下的30只sD大鼠于BDL模型建立4周后采用随机数字表法分为3组，动态观察给予生理盐水（生理盐水组）、内皮素-1（ET-1，ET-1组）、NO供体亚硝基半胱氨酸（GSNO，GSNO组）后BDL模型大鼠肝脏微循环的急性改变。组间比较采用单因素方差分析，组内比较采用配对样本t检验。结果BDL模型大鼠共死亡9只，存活率为85．0％（51／60）。假手术组大鼠全部存活。随着BDL造模时间延长，肝脏肝窦直径逐渐变小，BDL模型2周组、BDL模型4周组及BDL模型6周组肝窦直径分别为（13．6±1．0）μm、（8．8±0．7）μm和（8．0±0．5）μm，与假手术组的（17．4±1．0）μm比较，差异有统计学意义（t=5．86，18．24，15．57，P〈0．05）。随着BDL造模时间延长，肝窦开放数量逐渐减少，BDL模型2周组、BDL模型4周组及BDL模型6周组肝窦开放数量分别为（6．8±0．8）个／200μm、（4．3±1．8）个／200μm、（4．0±1．2）个／200μm，与假手术组的（8．8±0．5）／200μm比较，差异有统计学意义（t=3．25，2．77，2．12，P〈0．05）。注射生理盐水15min后，生理盐水组肝窦直径为（7．2±1．2）μm，与注射前的（6．9±0．5）Dm比较，差异无统计学意义（t=0．89，P〉0．05）；肝窦开放数量为（6．8±1．4）＋／200μm，与注射前的（6．6±0．4）个／200μm比较，差异无统计学意义（t=1．12，P〉0．05）。注射ET-115min后，ET-1组肝窦直径为（5．4±0．5）μm，与注射前的（7．9±0．6）μm比较，差异有统计学意义（t=7�Objective To investigate the value of intravital fluorescence microscopy in the observation of the changes of hepatic microcirculation in the rat model with hepatic cirrhosis and portal hypertension. Methods Seventy male SD rats were selected. According to the random number table, 40 SD rats were randomly divided into the sham operation group, bile duct ligation （BDL） 2 weeks group, 4 weeks group and 6 weeks group, there were 10 rats in each group, and the hepatic microcirculation of the rats was observed with intravital fluorescence microscope; the remaing 30 SD rats were randomly divided into the normal saline （ NS ） group, endothelin-1（ET-1） group and the S-nitrosoglutathion （GSNO） group at 4 weeks later after the establishment of BDL model. The changes of hepatic microcirculation of the 3 groups were observed. All data were analyzed using the one-way analysis of variance （ANOVA） or paired samples t test. Results Nine rats died in the BDL model groups, and the survival rate was 85.0% （51/60）. All rats in the sham operation group were survived. The hepatic sinusoid diameters were decreased as time passed by. The hepatic sinusoid diameters of the BDL 2 weeks group, 4 weeks group and 6 weeks group were （ 13.6 ± 1.0） μm, （8.8 ±0.7） μm and （8.0 ±0.5）μm, respectively, which were significantly shorter than （ 17.4± 1. 0） μm of the sham operation group （ t = 5.86, 18.24, 15.57, P 〈 0. 05 ）. The hepatic sinusoid densities of the BDL 2 weeks group, 4 weeks group and 6 weeks group were （6.8 ±0.8）/ 200 μm, （4.3 ± 1.8 ）/200 μm and （4.0 ± 1.2）/200μm, which were significantly lesser than （8.8 ± 0.5）/200 （ t = 3.25, 2. 77, 2. 12, P 〈 0.05 ）. At 15 minutes after injection of NS, the hepatic sinusoid diameter of the NS group was （7.2 ± 1.2） μm, which was significantly different from （ 6.9 ± 0.5 ）μm before injection of NS （ t = 0. 89, P 〉0.05） ; the hepatic sinusoid density of the NS group before and after injection of NS were （

关 键 词：肝硬化 门静脉高压 肝脏微循环 活体荧光显微镜 

分 类 号：R575.2[医药卫生—消化系统]