腹腔高壓對肝臟氧化還原狀態(tài)和組織病理變化的影響
發(fā)布時間：2018-07-22 15:28
【摘要】：目的：研究不同腹腔內(nèi)壓力升高和不同持續(xù)時間對肝功能和肝細胞的影響,為臨床研究腹內(nèi)高壓與多器官功能障礙的關(guān)系及其治療提供實驗依據(jù)。 方法：實驗用45只雄性SD (Sprague-Dawley)大鼠隨機分為三個組：對照組,腹內(nèi)壓力(intra-abdominal pressure, IAP)10mmHg模型組和IAP20mmHg模型組,每個組15只。實驗動物模型采用氮氣氣腹法來制作SD大鼠腹腔高壓的模型,對照組除不充入氮氣外,其余步驟與模型組一致。每組15只大鼠再隨機分為三個組,每個組5只分別在造模成功持續(xù)1h,2hs,4hs后處死。達到預定實驗時間后,迅速取腹主動脈血標本檢測大鼠血中天門冬氨酸轉(zhuǎn)氨酶(AST)、丙氨酸轉(zhuǎn)氨酶(ALT)含量；取大鼠左葉肝組織,部分作HE染色切片光鏡下觀察肝組織病理變化,部分用電鏡液固定并在透射電鏡下觀察肝細胞超微結(jié)構(gòu)；取大鼠右葉肝組織凍存,后制作成組織勻漿檢測超氧化物歧化酶(SOD)和還原型谷胱甘肽(GSH)的活性以及丙二醛(MDA)的含量。 結(jié)果：1.成功制作了腹腔高壓的SD大鼠模型。2.肝功能結(jié)果：IAP10mmHg4hs組血清AST、ALT含量均顯著高于IAP10mmHg1h組(p0.01)和IAP10mmHg2hs組(p0.05)；而IAP10mmHg2hs組與IAP10mmHg1h組相比無顯著差異。IAP20mmHg4hs組血清ALT、AST含量均顯著大于IAP20mmHg1h組(p0.01)和IAP20mmHg2hs組(p0.05); IAP20mmHg2hs組血清ALT和AST含量與相同IAP1h組相比也有明顯差異(均p0.05)。持續(xù)1h的模型組血清ALT和AST含量與對照組1h組比較無顯著差異。模型組2hs血清ALT、AST含量：IAP20mmHg2hs組血清ALT、 AST含量均顯著高于對照組2hs組(p0.01)和IAP10mmHg2hs組(p0.05)。模型組4hs血清ALT、AST含量變化：IAP20mmHg4hs組血清ALT. AST含量與對照組4hs組比較,有顯著差異(p0.01),與IAP10mmHg4hs組血清比較,AST有顯著差異(p0.05),而ALT的差異無統(tǒng)計學意義；IAP10mmHg4hs組ALT和AST含量與持續(xù)4hs的對照組比較,有明顯差異(p0.05)。3.肝組織勻漿SOD,MDA,GSH檢測結(jié)果：當IAP為10mmHg并持續(xù)1,2,4hs的SOD活性與正常對照組并持續(xù)1,2,4hs的SOD活性差異無顯著改變(均p0.05)；當IAP為20mmHg時,持續(xù)1h組的SOD活性與1h正常對照組的活性無顯著差異(p0.05),而持續(xù)2hs和4hs組的SOD活性分別與其對照組的活性有顯著差異(均p0.05)。各組之間肝組織勻漿中MDA含量均無顯著差異(p0.05)。與各自相同持續(xù)時間的對照組相比,IAP10mmHg2hs組、IAP10mmHg4hs組、IAP20mmHg1h組、IAP20mmHg2hs組以及IAP20mmHg4hs組的GSH活性顯著減少,其差異有統(tǒng)計學意義(均p0.05)。4.肝組織病理形態(tài)學結(jié)果：IAP10mmHg4hs組,光鏡下,少量肝細胞輕度水腫,中央靜脈區(qū)附近肝竇輕度腫脹,可見核腫脹空泡；電鏡下,可見部分肝細胞膜的纖維突起變鈍,胞質(zhì)內(nèi)線粒體膜基本完好,但線粒體嵴減少。IAP20mmHg1h組光鏡下,少量肝細胞輕度水腫,細胞膜邊界清晰度欠佳,肝竇輕度充血；電鏡下與IAP10mmHg4hs組相似；IAP20mmHg2hs組肝細胞的損傷范圍較前更廣。IAP20mmHg4hs組光鏡下,肝小葉結(jié)構(gòu)不整齊,可見部分細胞點狀壞死,并出現(xiàn)少量炎癥細胞浸潤；電鏡下,可見部分肝細胞膜褶皺,纖維突起消失,核膜完整性受損,核碎裂,胞質(zhì)內(nèi)線粒體腫脹,膜不清,線粒體嵴溶解。 結(jié)論：1.成功制作了動物模型,可復制。2.腹腔高壓可導致肝功能損傷：相同IAP時,肝功能損害隨持續(xù)時間增加而加重；相同持續(xù)時間時,肝功能損害隨IAP升高而加重。3.腹內(nèi)高壓會導致肝細胞的氧化應激損傷作用,特別是當IAP達到20mmHg及以上。4.一定程度的腹腔高壓在持續(xù)一定時間后會導致肝細胞病理形態(tài)學和細胞超微結(jié)構(gòu)的改變。
[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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R459.7
本文編號：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.
【學位授予單位】：廣西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R459.7
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