腹腔高压对肝脏氧化还原状态和组织病理变化的影响
发布时间：2018-07-22 15:28
【摘要】：目的：研究不同腹腔内压力升高和不同持续时间对肝功能和肝细胞的影响,为临床研究腹内高压与多器官功能障碍的关系及其治疗提供实验依据。 方法：实验用45只雄性SD (Sprague-Dawley)大鼠随机分为三个组：对照组,腹内压力(intra-abdominal pressure, IAP)10mmHg模型组和IAP20mmHg模型组,每个组15只。实验动物模型采用氮气气腹法来制作SD大鼠腹腔高压的模型,对照组除不充入氮气外,其余步骤与模型组一致。每组15只大鼠再随机分为三个组,每个组5只分别在造模成功持续1h,2hs,4hs后处死。达到预定实验时间后,迅速取腹主动脉血标本检测大鼠血中天门冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)含量；取大鼠左叶肝组织,部分作HE染色切片光镜下观察肝组织病理变化,部分用电镜液固定并在透射电镜下观察肝细胞超微结构；取大鼠右叶肝组织冻存,后制作成组织匀浆检测超氧化物歧化酶(SOD)和还原型谷胱甘肽(GSH)的活性以及丙二醛(MDA)的含量。 结果：1.成功制作了腹腔高压的SD大鼠模型。2.肝功能结果：IAP10mmHg4hs组血清AST、ALT含量均显著高于IAP10mmHg1h组(p0.01)和IAP10mmHg2hs组(p0.05)；而IAP10mmHg2hs组与IAP10mmHg1h组相比无显著差异。IAP20mmHg4hs组血清ALT、AST含量均显著大于IAP20mmHg1h组(p0.01)和IAP20mmHg2hs组(p0.05); IAP20mmHg2hs组血清ALT和AST含量与相同IAP1h组相比也有明显差异(均p0.05)。持续1h的模型组血清ALT和AST含量与对照组1h组比较无显著差异。模型组2hs血清ALT、AST含量：IAP20mmHg2hs组血清ALT、 AST含量均显著高于对照组2hs组(p0.01)和IAP10mmHg2hs组(p0.05)。模型组4hs血清ALT、AST含量变化：IAP20mmHg4hs组血清ALT. AST含量与对照组4hs组比较,有显著差异(p0.01),与IAP10mmHg4hs组血清比较,AST有显著差异(p0.05),而ALT的差异无统计学意义；IAP10mmHg4hs组ALT和AST含量与持续4hs的对照组比较,有明显差异(p0.05)。3.肝组织匀浆SOD,MDA,GSH检测结果：当IAP为10mmHg并持续1,2,4hs的SOD活性与正常对照组并持续1,2,4hs的SOD活性差异无显著改变(均p0.05)；当IAP为20mmHg时,持续1h组的SOD活性与1h正常对照组的活性无显著差异(p0.05),而持续2hs和4hs组的SOD活性分别与其对照组的活性有显著差异(均p0.05)。各组之间肝组织匀浆中MDA含量均无显著差异(p0.05)。与各自相同持续时间的对照组相比,IAP10mmHg2hs组、IAP10mmHg4hs组、IAP20mmHg1h组、IAP20mmHg2hs组以及IAP20mmHg4hs组的GSH活性显著减少,其差异有统计学意义(均p0.05)。4.肝组织病理形态学结果：IAP10mmHg4hs组,光镜下,少量肝细胞轻度水肿,中央静脉区附近肝窦轻度肿胀,可见核肿胀空泡；电镜下,可见部分肝细胞膜的纤维突起变钝,胞质内线粒体膜基本完好,但线粒体嵴减少。IAP20mmHg1h组光镜下,少量肝细胞轻度水肿,细胞膜边界清晰度欠佳,肝窦轻度充血；电镜下与IAP10mmHg4hs组相似；IAP20mmHg2hs组肝细胞的损伤范围较前更广。IAP20mmHg4hs组光镜下,肝小叶结构不整齐,可见部分细胞点状坏死,并出现少量炎症细胞浸润；电镜下,可见部分肝细胞膜褶皱,纤维突起消失,核膜完整性受损,核碎裂,胞质内线粒体肿胀,膜不清,线粒体嵴溶解。 结论：1.成功制作了动物模型,可复制。2.腹腔高压可导致肝功能损伤：相同IAP时,肝功能损害随持续时间增加而加重；相同持续时间时,肝功能损害随IAP升高而加重。3.腹内高压会导致肝细胞的氧化应激损伤作用,特别是当IAP达到20mmHg及以上。4.一定程度的腹腔高压在持续一定时间后会导致肝细胞病理形态学和细胞超微结构的改变。
[Abstract]:Objective: To study the effects of different intraperitoneal pressure and duration on liver function and liver cells, and to provide experimental basis for the clinical study of the relationship between intraperitoneal high pressure and multiple organ dysfunction.
Methods: 45 male SD (Sprague-Dawley) rats were randomly divided into three groups: the control group, the intra-abdominal pressure (IAP) 10mmHg model group and the IAP20mmHg model group, and each group 15. The experimental animal model was made by nitrogen pneumoperitoneum method to make the model of abdominal pressure in the peritoneal cavity of the SD rats, and the control group was not filled with nitrogen. The steps were the same as that in the model group. 15 rats in each group were randomly divided into three groups, and 5 rats in each group were executed after successful continuous 1H, 2hs, and 4hs. After the experiment time was reached, the blood samples from the abdominal aorta were quickly examined to detect the concentration of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the blood of rats. The pathological changes of liver tissue were observed under HE staining microscope. The ultrastructure of liver cells was fixed by electron microscope and observed under transmission electron microscope. The rat right lobe liver tissue was frozen, and then the activity of superoxide dismutase (SOD) and reduced glutathione (GSH) and the content of malondialdehyde (MDA) were detected by tissue homogenate.
Results: 1. the results of.2. liver function were successfully made in SD rat model of abdominal hypertension: the serum AST, ALT content in group IAP10mmHg4hs were significantly higher than that in group IAP10mmHg1h (P0.01) and IAP10mmHg2hs group (P0.05), but there was no significant difference between IAP10mmHg2hs and IAP10mmHg1h group. 1) and IAP20mmHg2hs group (P0.05), the serum ALT and AST content in IAP20mmHg2hs group were also significantly different from that of the same IAP1h group (P0.05). The serum ALT and AST content of the model group of continuous 1H was not significantly different from that of the control group. .01 and IAP10mmHg2hs group (P0.05). The change of ALT and AST content in the serum of model group: the AST content of serum ALT. in IAP20mmHg4hs group was significantly different from that of control group 4hs group (P0.01). In group comparison, there was a significant difference (P0.05).3. liver homogenate SOD, MDA, and GSH detection results: there was no significant difference between the SOD activity of IAP as 10mmHg and the normal control group and the SOD activity of the normal control group, and there was no significant difference between the viability of the 1,2,4hs and the activity of the normal control group. The activity of SOD in the continued 2hs and 4hs group was significantly different from that of the control group (P0.05). There was no significant difference in the MDA content in the liver homogenate between each group (P0.05). The GSH activity of the IAP10mmHg2hs group, IAP10mmHg4hs, IAP20mmHg1h, IAP20mmHg2hs and IAP20mmHg4hs group decreased significantly compared with the control group with the same duration. The difference was statistically significant (P0.05).4. hepatic histopathological results: group IAP10mmHg4hs, light microscopy, a small amount of liver cells with mild edema, mild swelling of the hepatic sinusoids near the central venous area, and nuclear swelling vacuoles; under electron microscopy, the fibrous protuberances of some hepatic cell membranes were blunt, and the mitochondria in the cytoplasm were basically intact, but mitochondria were basically intact, but mitochondria The crest reduced.IAP20mmHg1h group light microscopy, a small amount of liver cells with mild edema, poor clarity of the boundary of the cell membrane, mild hyperemia of the hepatic sinusoids, similar to that of the IAP10mmHg4hs group. The damage range of hepatocytes in group IAP20mmHg2hs was more extensive than that of group.IAP20mmHg4hs, the structure of liver lobule was not neat, some cells were punctiform necrosis, and a small amount of liver cells appeared. Inflammatory cells infiltrated; under electron microscope, some hepatic cell membrane folds, fibrous protuberances disappeared, nuclear membrane integrity damaged, nuclear fragmentation, mitochondria swelling in the cytoplasm, membrane indistinct, mitochondrial crista dissolving.
Conclusion: 1. the animal model was successfully made, and the liver function damage could be caused by the replication of.2. abdominal pressure. When the same duration of IAP, the liver function damage increased with the increase of duration. In the same duration, the liver function damage increased with the increase of IAP, which could lead to the oxidative stress damage of the liver cells, especially when IAP reached 20mmHg. And.4. above a certain degree of intra-abdominal hypertension will lead to changes in liver cell pathomorphology and cell ultrastructure after a certain period of time.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7
本文编号：2137908
[Abstract]:Objective: To study the effects of different intraperitoneal pressure and duration on liver function and liver cells, and to provide experimental basis for the clinical study of the relationship between intraperitoneal high pressure and multiple organ dysfunction.
Methods: 45 male SD (Sprague-Dawley) rats were randomly divided into three groups: the control group, the intra-abdominal pressure (IAP) 10mmHg model group and the IAP20mmHg model group, and each group 15. The experimental animal model was made by nitrogen pneumoperitoneum method to make the model of abdominal pressure in the peritoneal cavity of the SD rats, and the control group was not filled with nitrogen. The steps were the same as that in the model group. 15 rats in each group were randomly divided into three groups, and 5 rats in each group were executed after successful continuous 1H, 2hs, and 4hs. After the experiment time was reached, the blood samples from the abdominal aorta were quickly examined to detect the concentration of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the blood of rats. The pathological changes of liver tissue were observed under HE staining microscope. The ultrastructure of liver cells was fixed by electron microscope and observed under transmission electron microscope. The rat right lobe liver tissue was frozen, and then the activity of superoxide dismutase (SOD) and reduced glutathione (GSH) and the content of malondialdehyde (MDA) were detected by tissue homogenate.
Results: 1. the results of.2. liver function were successfully made in SD rat model of abdominal hypertension: the serum AST, ALT content in group IAP10mmHg4hs were significantly higher than that in group IAP10mmHg1h (P0.01) and IAP10mmHg2hs group (P0.05), but there was no significant difference between IAP10mmHg2hs and IAP10mmHg1h group. 1) and IAP20mmHg2hs group (P0.05), the serum ALT and AST content in IAP20mmHg2hs group were also significantly different from that of the same IAP1h group (P0.05). The serum ALT and AST content of the model group of continuous 1H was not significantly different from that of the control group. .01 and IAP10mmHg2hs group (P0.05). The change of ALT and AST content in the serum of model group: the AST content of serum ALT. in IAP20mmHg4hs group was significantly different from that of control group 4hs group (P0.01). In group comparison, there was a significant difference (P0.05).3. liver homogenate SOD, MDA, and GSH detection results: there was no significant difference between the SOD activity of IAP as 10mmHg and the normal control group and the SOD activity of the normal control group, and there was no significant difference between the viability of the 1,2,4hs and the activity of the normal control group. The activity of SOD in the continued 2hs and 4hs group was significantly different from that of the control group (P0.05). There was no significant difference in the MDA content in the liver homogenate between each group (P0.05). The GSH activity of the IAP10mmHg2hs group, IAP10mmHg4hs, IAP20mmHg1h, IAP20mmHg2hs and IAP20mmHg4hs group decreased significantly compared with the control group with the same duration. The difference was statistically significant (P0.05).4. hepatic histopathological results: group IAP10mmHg4hs, light microscopy, a small amount of liver cells with mild edema, mild swelling of the hepatic sinusoids near the central venous area, and nuclear swelling vacuoles; under electron microscopy, the fibrous protuberances of some hepatic cell membranes were blunt, and the mitochondria in the cytoplasm were basically intact, but mitochondria were basically intact, but mitochondria The crest reduced.IAP20mmHg1h group light microscopy, a small amount of liver cells with mild edema, poor clarity of the boundary of the cell membrane, mild hyperemia of the hepatic sinusoids, similar to that of the IAP10mmHg4hs group. The damage range of hepatocytes in group IAP20mmHg2hs was more extensive than that of group.IAP20mmHg4hs, the structure of liver lobule was not neat, some cells were punctiform necrosis, and a small amount of liver cells appeared. Inflammatory cells infiltrated; under electron microscope, some hepatic cell membrane folds, fibrous protuberances disappeared, nuclear membrane integrity damaged, nuclear fragmentation, mitochondria swelling in the cytoplasm, membrane indistinct, mitochondrial crista dissolving.
Conclusion: 1. the animal model was successfully made, and the liver function damage could be caused by the replication of.2. abdominal pressure. When the same duration of IAP, the liver function damage increased with the increase of duration. In the same duration, the liver function damage increased with the increase of IAP, which could lead to the oxidative stress damage of the liver cells, especially when IAP reached 20mmHg. And.4. above a certain degree of intra-abdominal hypertension will lead to changes in liver cell pathomorphology and cell ultrastructure after a certain period of time.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R459.7
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